@article {1185, title = {Acacia nilotica Pod Extract has an Anti-cancer Effect on the U937 Cell Line}, journal = {Pharmacognosy Research}, volume = {15}, year = {2023}, month = {October 2023}, pages = {771-775}, type = {Original Article}, chapter = {771}, abstract = {

Background: Acacia nilotica (A. nilotica), a tree that thrives in tropical regions, has been reported to have various medicinal uses and various pharmacological properties. Potential anti-cancer effects are one of the medical properties of parts of this tree. Objectives: To explore and evaluate the cytotoxic effect of A. nilotica, we used Acacia nilotica{\textquoteright}s pod extract on U937 cell lines as an acute myeloid leukemia model. Materials and Methods: Microscopic analysis, flow cytometry, and WST-1 tests were performed on the U937 cell line after 72 hr of treatment with different concentrations of ethanol pod extract from Acacia nilotica (A. nilotica) to test for DNA cell cycle, cell viability and apoptosis, and to determine the IC50 of the drug. Results: Treatment with A. nilotica pod extract significantly induced cell death in U937-treated cells compared with untreated control cells. Microscopic analysis revealed morphological changes in dead and fragmented cells associated with a reduction in cell number. A reduction in cell viability with an increase of cells in the late apoptosis stage by flow cytometry indicated the cytotoxic effect of A. nilotica and accumulation of cells in the sub-G1 phase of the DNA cell cycle analysis. The IC50 was identified by the WST-1 assay. Conclusion: The findings of the present study suggest that A. nilotica may be employed as a natural anti-cancer agent in research on acute myeloid leukemia in humans.

}, keywords = {Apoptosis, cytotoxicity, Flow cytometry, Natural drugs}, doi = {10.5530/pres.15.4.081}, author = {Halla Falih Bakheit and Sebastien Taurin and Elwaleed Mohamed Elamin and Moiz Bakhiet} } @article {1133, title = {Essential Oils of Rosmarinus officinalis and Eucalyptus globulus Cultivated in the Mountainous Region of the State of Rio de Janeiro (Brazil): Chemical Profile and Antileishmanial Activity}, journal = {Pharmacognosy Research}, volume = {15}, year = {2023}, month = {February 2023}, pages = {297-306}, type = {Original Article }, chapter = {297}, abstract = {

The present study aimed to characterize the chemical profile of essential oils of Rosmarinus officinalis (EORo) and Eucalyptus globulus (EOEg) cultivated in the mountainous region of the state of Rio de Janeiro, Brazil, and to verify the leishmanicidal potential, as well as to determine the cytotoxicity in vitro in mammalian cells. EORo had α-Pinene (37.99\%), Cineol (21.48\%), D-Verbenone (5.78\%) and Limonene (3.28\%) as major constituents. The EOEg had as major components Cineol (75.52\%), Limonene (8.82\%) and o-Cymeno (7.81\%). The EOEg had a lower IC50 value of 14.03 {\textpm} 2.08 μg/mL, while EORo had an IC50 value of 31.12 {\textpm} 4.6 μg/mL, both during the 24-hr exposure period. In the 48-hr period, the IC50 value was almost twice as high with rosemary oil, while in eucalyptus oil there was no significant difference when exposed in this period. The selectivity of EOs was demonstrated in murine cells, showing almost 30 times less toxic to the cell than to the parasite. It can be concluded that this is a pioneering study on the potential of these essential oils against the leishmania parasite. Furthermore, the low toxicity effect on the host cell encourages future studies and in vivo applications.

}, keywords = {cytotoxicity, E. globulus, Essential oil, Leishmanicidal, R. officinalis}, doi = {10.5530/pres.15.2.032}, author = {Rog{\'e}ria Cristina Zauli and Tarc{\'\i}sio Martins Santos and Alberto Wisniewski Jr and Patr{\'\i}cia Xander and Aislan Cristina Rheder Fagundes Pascoal} } @article {1086, title = {A New Cytotoxic Ceramide from Dalbergia boehmii Taub. Stem Bark}, journal = {Pharmacognosy Research}, volume = {14}, year = {2022}, month = {October 2022}, pages = {442-445}, type = {Original Article}, chapter = {442}, abstract = {

Background: Dalbergia boehmii (Leguminosae), is a medicinal plant used in the Northen part of Cameroon, against digestive tract diseases and fever. Objectives: This work which is part of the search for bioactive molecules through natural products, aimed to isolate, characterize and evaluate the cytotoxicity of compounds from D. boehmii. Materials and Methods: Chemical investigations were carrying out on stem barks of Dalbergia boehmii using chromatographic techniques. Structures elucidation of isolated compounds were done using 1D and 2D NMR as well as EI-MS and literature data. Some isolated compounds were evaluated in vitro against MCF-7 cell lines for their cytotoxicity activity, using the MTT colorimetric method. Results: The chemical investigation leaded to the isolation of a new ceramide derivative named Dalbergiamide (1), a triterpene identified as olean-12-ene-3,11-dione (2) isolated here for the first time from Dalbergia genus, along with two known compounds, glucoside 3-O-β- D-sitosterol (3) and glucoside 3-O-β-D-stigmasterol (4). Compounds 1 and 2 were found to be cytotoxic at 50 μM with IC50 value of 17.13 μM and 35.52 {\textpm} 2.95 μM respectively. Conclusion: These promising results suggested more in-depth studies should be carried out on this plant in order to assess its potential in the fight against cancer

}, keywords = {cytotoxicity, Dalbergia boehmii, MCF-7, Natural Products}, doi = {10.5530/pres.14.4.64}, author = {Jean Pierre Abdou and Jean No{\"e}l Nyemb and Jean Momeni and Atikou Fadimatou and Abel Jo{\"e}l Yaya Gbaweng and Jean Paul Tsopmejio and Guidawa Fawa and Muhammad I. Choudhary} } @article {853, title = {Cytotoxic Essential Oil from Annona sengalensis Pers. Leaves}, journal = {Pharmacognosy Research}, volume = {2}, year = {2021}, month = {September 2010}, pages = {211-214}, type = {Original Article}, chapter = {211}, abstract = {

The cytotoxicity against brine shrimp of the essential oil obtained from the leaves of Annona senegalensis Pers. (Annonaceae) was studied. The confirmation of this toxicity has been done by using selected tumor cell lines (A549, HT29, MCF 7, RPMI, and U251). The results showed that the total oil and its fractions have showed mild to moderate cytotoxicity in brine shrimp lethality bioassay with LC50 = 27.3 μg/ml, and against some human tumor cell lines. The total oil and its fractions were analyzed by gas chromatography/mass spectroscopy (GC/MS). Seventy three compounds were identified.

}, keywords = {Annona senegalensis, Brine shrimp, cytotoxicity, Essential oil, GC/MS}, doi = {10.4103/0974-8490.69105}, author = {A L Ahmed, and S E M Bassemx and Y H Mohamed and MW Gamila} } @article {1002, title = {Effects of Extraction Solvents on Phytochemical Screening, Cytotoxicity and Anti-Obesity Activities of Selected Sri Lankan Medicinal Plants}, journal = {Pharmacognosy Research}, volume = {13}, year = {2021}, month = {October 2021}, pages = {246-256}, type = {Original Article}, chapter = {246}, abstract = {

Background: Phytoconstituents are known to have an arena of health benefits. The choice of extraction solvents for a desired pharmacological activity is poorly characterized. Objectives: The present study compared the use of different solvents to extract phytoconstituents, evaluate cytotoxicity and anti-obesity activities. Materials and Methods: Leaves, bark, flowers or tubers of fourteen commonly utilized plants in the Sri Lankan Ayurvedic system were used to prepare extracts of distilled water (DW), ethanol or n-hexane. Phytochemical screening, total phenolic and flavonoid contents (TPC and TFC), cytotoxicity and triglyceride assays were performed. Results: DW served as the best solvent to extract the highest yield of phytochemicals followed by ethanol marking hexane as the least efficient solvent. The water extract (WE) of Averrhoa bilimbi leaves and ethanol extract (EE) of Cassia auriculata showed the highest TPC. The TFC was lowest in the hexane extract (HE) of all plants. The WE of plants showed >=75\% cell viability below 500 μg/mL with no potential anti-adipogenic activity. The EE of Justicia adhathoda showed maximum lipid inhibition (88.31\%), followed by Aerva lanata (74.65\%), and Cyperus rotundus (74.04\%) at 100 μg/mL. The EE of Ipomoea aquatica (45.11\%), Murrya koenigii (41.04\%), and Hemidesmus indicus (32.97\%) showed the highest viable anti-adiposity activity at 50 μg/mL. Conclusion: The WE of most plants provided a greater yield of phytoconstituents enabling their safe usage in higher doses during Ayurvedic medication. Ethanol served as a better solvent to extract constituents responsible for anti-adipogenesis and no possible correlation was found between the phenolic content and anti-adiposity.

}, keywords = {3T3-L1, Anti-adipogenesis, Ayurveda, cytotoxicity, Medicinal plants, Phytochemicals}, doi = {10.5530/pres.13.4.18}, author = {Silva Waruni Randika and Nugara Ruwani Nilushi and Manage Pathmalal and Undugoda Lanka and Udayanga Dhanushka and Liyanage Renuka Nilmini and Galhena Prasanna} } @article {179, title = {Evaluation of the Antioxidant and Gastric Antiulcerogenic Activities of the Hydroalcoholic Extract and Leaf Fractions of Solanum stipulaceum Roem. and Schult.}, journal = {Pharmacognosy Research}, volume = {12}, year = {2021}, month = {August 2020}, pages = {328-335}, type = {Original Article}, chapter = {328}, abstract = {

Background: An earlier ethnobotanical study showed that the species Solanum stipulaceum was indicated first in many orders for the treatment of gastric ulcers. Aims: The aim of this study was to evaluate phytochemical profile and the antioxidant, antiulcerogenic, protective redox, and cytotoxicity activities of the hydroethanol extract (HEE) and leaf fractions of this plant. Materials and Methods: The HEE was obtained by maceration and fractionated by liquid{\textendash}liquid extraction in fractions: hexane fraction, chloroform fraction (CF), ethyl acetate fraction (EAF), and hydromethanol fraction (HMF). Phytochemical profile was evaluated by colorimetric methods. Antioxidant activity was measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The cellular toxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In vivo antiulcerogenic activity was investigated using the ethanol-induced acute ulcer model. Afterward, protective redox activity was assessed with the plasma and liver of these animals using the thiobarbituric acid reactive substances assay. Results: Tannins, free steroids, triterpenes, alkaloids, and flavonoids were found in HEE, and total flavonoid measured was 5.26 {\textpm} 0.23 μg/mg of the extract. The DPPH activity was higher than 70\% for HEE, EAF, and CF. Treatment with HEE (200 and 400 mg/kg) and CF, EAF, and HMF (200 mg/kg) inhibited the ulcer. CF reduces gastric lesions by 88.29\% (50 mg/kg) and 85.65\% (100 mg/kg). The HEE (400 mg/kg), CF, and EAF (200 mg/kg) reduced oxidative stress in the plasma and in the liver. HMF (200 mg/kg) reduced oxidative stress only in the liver. The HEE and fractions 10{\textendash}50 μg/mL (P \< 0.05) were nontoxic, maintaining cell viability at 65\%. Conclusion: The extract and fractions have antioxidant activity in vitro and ex vivo, antiulcerogenic effect in vivo, and low cytotoxicity up to 50 μg/mL.

}, keywords = {Antioxidant activity, Antiulcerogenic activity, cytotoxicity, Phytochemical screening, Solanum stipulaceum, Total flavonoid}, doi = {10.4103/pr.pr_2_20}, author = {Cirlane Alves Araujo de Lima and Daniela Feitosa de Oliveira and Andressa da Silva and Robson Silva de Lima and Jymmys Lopes dos Santos and Ant{\^o}nio Santos Dias and Andr{\'e} Luiz Lima Menezes dos Santos and Jamylle Nunes de Souza Ferro and Andrea Yu Kwan Villar Shan and Brancilene Santos de Araujo and Emiliano de Oliveira Barreto and Maria Silene da Silva and Josemar Sena Batista and Sara Maria Thomazzi and Sonia Piacente and Cosimo Pizza and Charles dos Santos Estevam} } @article {985, title = {Trypanocidal Activity of Traditional Antiparasitic Medicinal Plants from the Amazon}, journal = {Pharmacognosy Research}, volume = {13}, year = {2021}, month = {October 2021}, pages = {227-232}, type = {Original Article}, chapter = {227}, abstract = {

Background: Neglected Infectious Diseases control is an urgent need in endemic areas, but it{\textquoteright}s not a priority in the commercial interests of the pharmaceutical industry. Chagas disease (caused by Trypanosoma cruzi) is the parasitic disease of greatest impact in Latin America. Medicinal plants continue to be an affordable front-line option in endemic areas. Thus, we aimed to advance in the pharmacological evaluation of four medicinal plants traditionally used in the Amazon against parasitic infections. Materials and Methods: The plants were collected in the Amazon region of Colombia. The dry plant material was submitted to water percolation extraction. Extracts were tested in vitro against Trypanosoma cruzi (epimastigotes), and cytotoxicity was assessed against HepG2 and MRC5 cells. Finally, the general profile of metabolites present in the extracts was studied by thin-layer chromatography. Results: In vitro, against T. cruzi, extracts of Ambelania duckei and Curarea toxicofera shows concentration- dependent inhibition (IC50 of 221+/-29 and 50+/-5 μg/mL respectively), comparable with benznidazole (IC50: 0.7 μg/mL); while Abuta grandifolia and Aspidosperma excelsum exhibited IC50{\textquoteright}s \> 500 μg/mL. All extracts showed no cytotoxicity against HepG2 and MRC5 cells. Yields of extraction were between 3.2 and 9.5\% and preliminary phytochemical profile showed flavonoids and steroids in all extracts. Conclusion: Promising plants, traditionally used to treat other protozoan infections, could be assessed against T. cruzi. C. toxicofera exhibits good activity against epimastigotes of T. cruzi, being a species that can reasonably be considered for bioassay-guided antitrypanosomal fractionation.

}, keywords = {Chagas disease, cytotoxicity, Medicinal plants, Neglected Infectious Diseases, Trypanocidal agents, Trypanosoma cruzi}, doi = {10.5530/pres.13.4.15}, author = {Mar{\'\i}a-Helena Arias and Gustavo A. Vallejo and Giovanny Garavito} } @article {162, title = {Evaluation of Cytotoxicity and Antioxidant Potential of Bael Leaf (Aegle marmelos) on Human Hepatocellular Carcinoma Cell Line}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {August 2020}, pages = {267-271}, type = {Original Article}, chapter = {267}, abstract = {

Background: Aegle marmelos (Bael) is one of the important medicinal plants for curing human diseases. Nevertheless, the information about the cytotoxic and antioxidant effects of Bael leaf is not much. Objectives: The objective of this study is to evaluate the cytotoxicity and antioxidant activity of Bael leaf in human hepatocellular carcinoma (HepG2) cell lines. Materials and Methods: Bael leaves extract was prepared by 80\% (v/v) aqueous ethanol and measured for total phenolic compound and antioxidant capacity. The ability of ethanolic Bael leaf extract to inhibit hepatic cancer cells (HepG2 cell) growth was evaluated by determining the percentages of cell viability. The levels of intracellular reactive-oxygen species (ROS) and lipid peroxidation were determined. Results: Bael leaf extract demonstrated the potentiality to inhibit the cancer cell growth with a 50\% inhibitory concentration (IC50) at 50 μg/ml and 72 μg/ml after 24 h and 48 h incubated times, respectively. The total phenolic content and antioxidant capacity of ethanolic extract were 218.33 {\textpm} 43.81 mg gallic acid equivalent/g of dry extract and 33.62 {\textpm} 3.07 mg Trolox equivalent antioxidant capacity/g of dry extract, respectively. At the lowest concentration of Bael leaf extract, it was found that Bael leaf extract shows the antioxidant activity by the ability to lowering the level of ROS and malondialdehyde in HepG2 cell. Conclusion: Bael leaf has a high antioxidant component, which is beneficial and can be developed as new therapeutic uses. However, further studies on the benefits of Bael leaf should be performed for better realizing and effective use soon.

}, keywords = {Antioxidant, Bael, cytotoxicity, Human hepatocellular carcinoma cell line, Malondialdehyde, Reactive-oxygen species}, doi = {10.4103/pr.pr_15_20}, author = {Kanokwan Kulprachakarn and Sakaewan Ounjaijean and Somdet Srichairatanakool and Duangta Kanjanapothi} } @article {159, title = {Screening of Cytotoxic Activity of Hematite (α-Fe2O3) Nanoparticles from Butea monosperma on MCF-7 Cells}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {August 2020}, pages = {260-266}, type = {Original Article}, chapter = {260}, abstract = {

Background: Hematite (α-Fe2O3) has been used as an antimicrobial and disinfectant agent. Nevertheless, there are limited data about antitumor potential. This study was focused on investigating cytotoxic effects of Hematite (α-Fe2O3) from Butea monosperma flower extract on MCF -7 breast cancer cells and its mechanism of action. Materials and Methods: Thus, a green method was created for the synthesis of Hematite (α-Fe2O3) using an aqueous extract of B. monosperma flower. Synthesis of Hematite (α-Fe2O3) was described by different analytical techniques including ultraviolet-visible spectrophotometer, field-emission scanning electron microscopy, X-ray diffraction, and Fourier transforms infrared spectroscopy. Cell viability was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Reactive oxygen species (ROS) formation was measured using probe 2{\textquoteright},7{\textquoteright}-dichlorofluorescein diacetate and intracellular calcium (Cai2+) was evaluated with probe flu3-AM. Cells were treated with different concentrations of Hematite (α-Fe2O3) (1, 3, 6, 10, 15, 25, 50, and 100 μg/mL). Results: The results showed that Hematite (α-Fe2O3) hindered cell growth in a dose-dependent manner. Hematite (α-Fe2O3) appeared to have dose-dependent cytotoxicity against MCF-7 cells through activation of the ROS generation and an increase in the intracellular Cai2+ (half-maximal inhibitory concentration 52 {\textpm} 3.14). Conclusion: In conclusion, the results of this preliminary study demonstrated that Hematite (α-Fe2O3) from B. monosperma flower extract may be a potential therapeutic potential medicament for human breast cancer treatment.

}, keywords = {Butea monosperma, cytotoxicity, Hematite (α-Fe2O3), MCF-7 cell line, Nanoparticle}, doi = {10.4103/pr.pr_115_19}, author = {Debasish Pradhan and Shaktiprasad Pradhan and Bandana Behera and Adyasa Samantaray} } @article {201, title = {Toxicity Evaluation of Camellia sinensis var. assamica and Its Fermented Miang Product}, journal = {Pharmacognosy Research}, volume = {12}, year = {2020}, month = {January 2021}, pages = {430-436}, type = {Original Article}, chapter = {430}, abstract = {

Introduction: Camellia sinensis var. assamica (Assam tea or Cha-Miang [CM]) is widely accepted to be beneficial to health. The tea leaf is used to produce various fermented tea products such as black tea (China, India) and Miang (Thailand). Despite its medicinal properties, toxicological information regarding certain tea variety and its fermented Miang product, especially its long-term toxicity, is currently limited. This study aimed to evaluate the potential toxicity of the extract of fresh resh Cha-Miang leaves (CM) and its fermented Miang product (FCM) in both in vitro and in vivo models. Materials and Methods: Cytotoxic effect on cell viability of HepG2, HEK293, and EA.hy926 cell lines incubated with CM or FCM extract for 24 h and 48 h was investigated by the MTT assay. After that, 14-day repeat oral toxicity test was performed on Wistar rats. Results: No in vitro cytotoxic effect of CM and FCM extract was found in the tested cell types, at a dose up to 1000 {\textmu}g/ml. For in vivo study, both CM and FCM extracts at a dose of 300 mg/kg/day did not produce any sign or symptom of toxicity; no mortality was observed. Furthermore, investigation of hematological parameters, blood chemistry, body weight, and organ weight in the treated rats revealed no significant difference compared with that of normal controls. Conclusion: The results suggested that crude extract of CM and FCM at the doses used in this study are safe, however, further evaluation of possible chronic toxicity is recommended.

}, keywords = {Camellia sinensis var. assamica, cytotoxicity, Fermented tea product, Miang, Subacute toxicity}, doi = {10.4103/pr.pr_22_20}, author = {Sukanya Chachiyo and Kanokwan Kulprachakarn and Chalermpong Saenjum and Kittipan Rerkasem and Somdet Srichairatakool and Kongsak Boonyapranai and Wason Parklak and Voravuth Somsak and Sakaewan Ounjaijean} } @article {140, title = {Cytotoxicity and Apoptosis Induction of Sea Cucumber Holothuria atra Extracts}, journal = {Pharmacognosy Research}, volume = {11}, year = {2019}, month = {February 2019}, pages = {41-46}, type = {Original Article}, chapter = {41}, abstract = {

Context: Cancer is one of the major causes of death. Sea cucumbers are marine invertebrate that is widely used and utilized in traditional medicine. One of the most studied of sea cucumbers efficacy is its anticancer property. The anticancer property of sea cucumbers is much related to the content of an active compound called saponin. Objective: The objective of this research is to determine the cytotoxicity and apoptosis induction of sea cucumber Holothuria atra ethanol extract and to study its anticancer active compound. Materials and Methods: Sea cucumber of H. atra was taken from Halmahera waters, North Maluku Indonesia. The extraction was conducted by maceration method using ethanol 96\% then continued with liquid-liquid partition and separation using C18 column. The investigation of active compound content was conducted by phytochemical and spectroscopic analysis using liquid chromatography-ion trap-time of a flight-mass spectrophotometer (LC-IT-ToF-MS). The cytotoxicity test was performed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide method against human breast ductal carcinoma cell (T47D), human breast adenocarcinoma (MCF7), human colorectal adenocarcinoma cell (WiDr), and human cervix adenocarcinoma cell lines (HeLa). The apoptotic induction test was performed using flow cytometry using Fluorescein isothiocyanate (FITC) Annexin-V-Apoptosis and caspase-3 detection using FITC Active Caspase-3 kit. Results: Phytochemical test showed that ethanol extract of H. atra contained alkaloids, flavonoids, steroids-triterpenoids, phenols, and saponins. The extract showed cytotoxic activity against the 4 cell lines with Inhibition concentration 50 values ranging from 9.6 to 14.3 μg/ml. Flow cytometry analysis showed that the T47D cell population underwent apoptosis after treated with ethanol extract. The extract also activated caspase-3 on the T47D cells. The results of LC-IT-ToF-MS analysis showed that the active fraction from C18 column separation contained saponin and identified as Cucumechinol and Philinopgenin B. Conclusions: The results of this study indicated that H. atra active extract has good cytotoxicity and has potential to be developed as an anticancer agent.

}, keywords = {Apoptosis, caspase-3, cytotoxicity, Holothuria atra, Saponin, Sea cucumber}, doi = {10.4103/pr.pr_3_18}, author = {Muhammad Nursid and Endar Marraskuranto and Ekowati Chasanah} } @article {284, title = {Cytotoxicity and Apoptosis Induction of Sea Cucumber Holothuria atra Extracts}, journal = {Pharmacognosy Research}, volume = {11}, year = {2019}, month = {February,2019}, pages = {41-46}, type = {Original Article}, chapter = {41}, abstract = {

Context: Cancer is one of the major causes of death. Sea cucumbers are marine invertebrate that is widely used and utilized in traditional medicine. One of the most studied of sea cucumbers efficacy is its anticancer property. The anticancer property of sea cucumbers is much related to the content of an active compound called saponin. Objective: The objective of this research is to determine the cytotoxicity and apoptosis induction of sea cucumber Holothuria atra ethanol extract and to study its anticancer active compound. Materials and Methods: Sea cucumber of H. atra was taken from Halmahera waters, North Maluku Indonesia. The extraction was conducted by maceration method using ethanol 96\% then continued with liquid-liquid partition and separation using C18 column. The investigation of active compound content was conducted by phytochemical and spectroscopic analysis using liquid chromatography-ion trap-time of a flight-mass spectrophotometer (LC-IT-ToF-MS). The cytotoxicity test was performed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide method against human breast ductal carcinoma cell (T47D), human breast adenocarcinoma (MCF7), human colorectal adenocarcinoma cell (WiDr), and human cervix adenocarcinoma cell lines (HeLa). The apoptotic induction test was performed using flow cytometry using Fluorescein isothiocyanate (FITC) Annexin-V-Apoptosis and caspase-3 detection using FITC Active Caspase-3 kit. Results: Phytochemical test showed that ethanol extract of H. atra contained alkaloids, flavonoids, steroids-triterpenoids, phenols, and saponins. The extract showed cytotoxic activity against the 4 cell lines with Inhibition concentration 50 values ranging from 9.6 to 14.3 μg/ml. Flow cytometry analysis showed that the T47D cell population underwent apoptosis after treated with ethanol extract. The extract also activated caspase-3 on the T47D cells. The results of LC-IT-ToF-MS analysis showed that the active fraction from C18 column separation contained saponin and identified as Cucumechinol and Philinopgenin B. Conclusion: The results of this study indicated that H. atra active extract has good cytotoxicity and has potential to be developed as an anticancer agent.

}, keywords = {Apoptosis, caspase-3, cytotoxicity, Holothuria atra, Saponin, Sea cucumber.}, doi = {10.4103/pr.pr_3_18}, author = {Muhammad Nursid and Endar Marraskuranto and Ekowati Chasanah} } @article {288, title = {Antioxidant and Cytotoxic Potential of Endophytic Fungi Isolated from Medicinal Plant Tragia involucrata L.}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {April 2018}, pages = {188-194}, type = {Original Article}, chapter = {188}, abstract = {

Objective: The aim of the present study was to test the antioxidant and cytotoxic and DNA protection activities of ethyl acetate extracts of endophytic fungi isolated from Tragia involucrata Linn. (Euphorbiaceae). Materials and Methods: The 1, 1-diphenyl-2-picrylhydrazyl scavenging, reducing power, and total antioxidant assay were used to evaluate the antioxidant activity. Cytotoxic activity of endophytic fungal extracts against MCF-7 and MOLT-4 cell lines was carried out using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide method. Results: We have successfully isolated four endophytic fungi from T. involucrata, namely Penicillium citrinum CGJ-C1 (GenBank accession No. KT780618), P. citrinum CGJ-C2 (GenBank accession No. KP739821), Cladosporium sp. CGJ-D1 (GenBank accession No. KP739822), and Cryptendoxyla hypophloia CGJ-D2 (GenBank accession No. KT780619). The ethyl acetate extract of P. citrinum CGJ-C2 showed the highest antioxidant as well as cytotoxic activity among the four fungal extracts taken for the study. All the extracts showed moderate DNA protection activity. Conclusions: Further studies on the isolation and purification of the lead molecule will help in designing the novel therapy for different ailments associated with free radical generation.

}, keywords = {Antioxidants, cytotoxicity, Endophytes, Tragia involucrata}, doi = {10.4103/pr.pr_137_17}, author = {Ananda Danagoudar and Chandrashekhar G. Joshi and Sunil Kumar Ravi and Heggodu G. Rohit Kumar and Balenahalli Narasingappa Ramesh} } @article {215, title = {Cytotoxic Compounds from Wrightia pubescens (R.Br.)}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {February 2018}, pages = {9-15}, type = {Original Article}, chapter = {9}, abstract = {

Background: Mixtures of ursolic acid (1) and oleanolic acid (2) (1:1 and 1:2), oleanolic acid (2), squalene (3), chlorophyll a (4), wrightiadione (5), and α-amyrin acetate (6) were isolated from the dichloromethane (CH2Cl2) extracts of the leaves and twigs of Wrightia pubescens (R.Br.). Objectives: To test for the cytotoxicity potentials of 1{\textendash}6. Materials and Methods: The antiproliferative activities of 1{\textendash}6 against three human cancer cell lines, breast (MCF-7) and colon (HT-29 and HCT-116), and a normal cell line, human dermal fibroblast neonatal (HDFn), were evaluated using the PrestoBlue{\textregistered} cell viability assay. Results: Compounds 4, 1 and 2 (1:2), 2, 1 and 2 (1:1), and 5 exhibited the most cytotoxic effects against HT-29 with half maximal inhibitory concentration (IC50) values of 0.68, 0.74, 0.89, 1.70, and 4.07 μg/mL, respectively. Comparing 2 with its 1:1 mixture with 1 (IC50 = 1.70 and 7.18 μg/mL for HT-29 and HCT-116, respectively) and 1:2 mixture with 1 (0.74 and 3.46 μg/mL for HT-29 and HCT-116, respectively), 2 also showed strong cytotoxic potential against HT-29 and HCT-116 (0.89 and 2.33 μg/mL, respectively). Unlike the mixtures which exhibited low effects on MCF-7 (IC50 = 20.75 and 30.06 μg/mL for 1:1 and 1:2, respectively), 2 showed moderate activity against MCF-7 (10.99 μg/mL). Compound 6 showed the highest cytotoxicity against HCT-116 (IC50 = 4.07 μg/mL). Conclusion: Mixtures of 1 and 2 (1:1 and 1:2), 2, 3, 4, 5, and 6 from the CH2Cl2 extracts of the leaves and twigs of W. pubescens (R.Br.) exhibited varying cytotoxic activities. All the compounds except 6 exhibited the strongest cytotoxic effects against HT-29. On the other hand, 6 was most cytotoxic against HCT-116. Overall, the toxicities of 1{\textendash}6 were highest against HT-29, followed by HCT-116 and MCF-7. All the compounds showed varying activities against HDFn (IC50 \<30 μg/mL).

}, keywords = {Apocynaceae, Chlorophyll a, cytotoxicity, Half maximal inhibitory concentration, HCT-116, HDFn, HT-29, MCF-7, Oleanolic acid, PrestoBlue{\textregistered} cell viability assay, Squalene, Ursolic acid, Wrightia pubescens R. Brown, Wrightiadione, α-amyrin acetate}, doi = {10.4103/pr.pr_45_17}, author = {Mariquit M. De Los Reyes and Glenn G. Oyong and Vincent Antonio S. Ng and Consolacion Y. Ragasa} } @article {285, title = {High-Performance Liquid Chromatography-Fingerprint Analyses, In vitro Cytotoxicity, Antimicrobial and Antioxidant Activities of the Extracts of Two Cestrum Species Growing in Egypt}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {April 2018}, pages = {179-180}, type = {Original Article}, chapter = {173}, abstract = {

Background: Cancer diseases and microbial resistance are serious health disorders associated with oxidative stress and infectious diseases. Their risks can be reducing via using polyphenols-rich plants. Methodology: Different solvent extracts from two Cestrum species (Cestrum nocturnum and Cestrum elegans) were evaluated for their biological and chemical activities. Also, the chemical profiles of the most promising extracts were investigated via high-performance liquid chromatography (HPLC)-fingerprint analyses. Results: The tested extracts showed weak to moderate cytotoxicity against Vero cell line with IC50 values ranged from 133.67 μg/ ml to 57.634 μg/ml. The only noncytotoxic extractive fraction was the dichloromethane extract of C. elegans leaves with an IC50 value of 204.732 μg/ml, while the most toxic extract was the ethyl acetate extract of C. elegans flowers with an IC50 value of 19.22 μg/ml. The antimicrobial activity results revealed that the n-BuOH extract of C. nocturnum was the most active against four tested microbial strains with inhibition zones (10{\textendash}13 mm). Also, the water and n-BuOH extracts of C. elegans leave exhibited moderate activities with inhibition zones (7{\textendash}9 mm), while for C. elegans flowers both of water and methanol extracts showed strong activities (9{\textendash}14 mm). In the 2,2{\textquoteright}-diphenyl-1-picrylhydrazyl assay, the most active fraction was EtOAc with IC50 values of 100.52 μg/ml and 64.40 μg/ml for C. elegans leaves and flowers respectively, while for C. nocturnum the most active fraction was methanol with an IC50 value of 161.16 μg/ml, all relative to 7.60 μg/ml of ascorbic acid. HPLC-fingerprint analyses revealed that the major identified compounds in the ethyl acetate extract of C. elegans flowers are caffeic acid, coumaric acid, vanillin, and rutin, while for the n-butanol extract of C. nocturnum leaves are coumaric acid and vanillin. Conclusion: The obtained results revealed that the two species can be used as natural sources of antioxidant compounds with low cytotoxic effect on the mammalian cell line.

}, keywords = {2, 2{\textquoteright}-Diphenyl-1-picrylhydrazyl, Antimicrobial, Cestrum species, cytotoxicity, high-performance liquid chromatography-fingerprint analyses, Solanaceae}, doi = {10.4103/pr.pr_145_17}, author = {Sami M. Nasr and Mosad A. Ghareeb and Mona A. Mohamed and Nehal M. Elwan and Abd El-Wanes Anter Abdel-Aziz and Mohamed S. Abdel-Aziz} } @article {360, title = {HPLC-DAD-ESI-MS/MS Characterization of Bioactive Secondary Metabolites from Strelitzia nicolai Leaf Extracts and Their Antioxidant and Anticancer Activities In vitro}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {October 2018}, pages = {368-378}, type = {Original Article}, chapter = {368}, abstract = {

Background: Strelitzia nicolai Regel and K{\"o}rn (Strelitziaceae) is native to Southern Africa whose phytochemistry and pharmacology were slightly investigated. Materials and Methods: In the current work, different solvent extracts of S. nicolai were screened for their chemical profiles through high-performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS/MS) analyses. Furthermore, their in vitro antioxidant, cytotoxic, and anticancer activities were evaluated using 2,2{\textquoteright}-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2{\textquoteright}-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) \& ferric reducing antioxidant power (FRAP) and crystal violet staining (CVS) colorimetric assays, respectively. Results: HPLC-DAD-ESI-MS/MS analyses led to the identification of nineteen and eleven phenolic compounds from the ethyl acetate and n-butanol extracts, respectively including flavonoids (e.g., quercetin 3-(2 G-rhamnosylrutinoside, quercetin, quercetin-3-O-glucoside, kaempferol-3,7-O-dirhamnoside, isorhamnetin-3-O-rutinoside and kaempferol-3-O-glucoside), phenolic acids derivatives (e.g., chlorogenic acid glycoside, protocatechuic acid-O-glucoside and caftaric acid ), chalcones (e.g., xanthoangelol), and phenylethanoids (e.g., ligstroside glucoside). Moreover, in the DPPH assay the IC50 value of the most active ethyl acetate extract was 20.49 μg/mL, relative to 2.92 μg/mL of ascorbic acid. ABTS and FRAP results reinforced the results of DPPH assay. According to the National Cancer Institute criteria, the tested extracts showed weak to moderate cytotoxic activities with IC50 values ranged from 65.23 to 451.29 μg/ mL. Furthermore, the EtOAc and n-BuOH extracts showed a noticeable anticancer activity with CVS spectroscopic readings for liver hepatocellular carcinoma growth 0.806 and 0.684 at a concentration (125 μg/mL), as well as 0.730 and 0.618 at concentration (500 μg/mL), respectively against control at 1.022. Conclusion: The obtained results reveal the high efficacy of the phenolic-rich extracts from S. nicolai as naturally occurring antioxidant and anti-tumor agents.

}, keywords = {Anticancer activity, Antioxidant activity, cytotoxicity, HPLC-DAD-ESI-MS/MS, Polyphenolics, Strelitzia nicolai}, doi = {10.4103/pr.pr_89_18}, author = {Mosad Ghareeb and Amal Saad and Wafaa Ahmed and Laila Refahy and Sami Nasr} } @article {420, title = {In vitro Assessment of Antioxidant, Antimicrobial, Cytotoxic, Anti-inflammatory, and Antidiabetic Activities of Campanula retrorsa Crude Extracts}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {October 2018}, pages = {397-403}, type = {Original Article}, chapter = {397}, abstract = {

Background: Campanula retrorsa belongs to the Campanulaceae family and the Campanula genus. The species of this latter are traditionally known for their antiallergic, spasmolytic, antiphlogistic, antioxidant, and antiviral properties. Few studies were realized on the bioactivity of Calicotome villosa crude extracts. Objective: The objective was to investigate, for the first time, the antioxidant, antimicrobial, cytotoxic, antidiabetic, and anti-inflammatory potentials of dichloromethane, methanol, and aqueous crude extracts of C. retrorsa leaves, flowers, and stems. Materials and Methods: The antioxidant activity of different crude extracts was determined using the free radical, 1, 1-diphenyl-2-picrylhydrazyl. Antimicrobial activity against three Gram-positive (Staphylococcus aureus, Bacillus cereus, and Enterococcus faecium), two Gram-negative (Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumanii) bacterial strains and a fungal strain (Candida albicans) was evaluated in a plate-hole diffusion assay. Cytotoxic activity of C. retrorsa dichloromethane and methanol crude extracts on Hela cells was realized using the Trypan blue dye exclusion technique. Antidiabetic activity against α-glucosidase and anti-inflammatory activity against bee venom phospholipase A2 (PLA2) were assessed using a colorimetric test. Results: Leaf and flower aqueous extracts possessed very high radical scavenging activity = 90\%. The dichloromethane extracts of leaves and flowers were the most active against A. baumanii and C. albicans. After 48 h of treatment, all extracts except the methanol flower extract showed an inhibition of 90\% of cell growth at 2 mg/mL. Dichloromethane and methanol stem crude extracts showed a modest inhibition of α-glucosidase. While no inhibition of the PLA2 was displayed by any of the tested extracts. Conclusion: Obtained results give an overall view on C. retrorsa crude extracts{\textquoteright} bioactivities.

}, keywords = {Antidiabetic activity, Bioactivity, Campanula retrorsa, Crude extracts, cytotoxicity}, doi = {10.4103/pr.pr_73_18}, author = {Josiane Alhage and Hoda Elbitar and Samir Taha and Thierry Benvegnu} } @article {279, title = {Phytochemical Screening, Total Phenolic Content, Antioxidant, and Cytotoxic Activity of Five Peruvian Plants on Human Tumor Cell Lines}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {April 2018}, pages = {161-165}, type = {Original Article}, chapter = {161}, abstract = {

Background: Cancer is considered a serious public health problem in the world; searching alternative treatments of medicinal plants constitute a promissory field to find new anticancer drugs. Objective: The aim of this study is to evaluate the phytochemical screening, total phenolic content (TPC), antioxidant, and cytotoxic activity of ethanol extracts of Waltheria ovata (WO), Piper aduncum (PA), Myrciaria dubia (MD), Physalis peruviana (PP), and Geranium dielsianum (GD) on human tumor cell lines. Materials and Methods: Phytochemical screening was assessed using chemical reactives. TPC was developed using Folin Ciocalteu reactive, and the antioxidant activity was determined against 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals by spectrophotometry. The cytotoxic activity was determined on human tumor cell lines followed as: MCF-7, H-460, HT-29, M-14, K-562, and DU-145. Results: Phytochemical study confirmed flavonoids and phenolic compounds in all extracts. TPC was found to be the highest in WO extract (1250 mg of gallic acid equivalent/g of dried extract) rather than other extracts. The highest antioxidant activity was stablished in WO extract for DPPH and ABTS radical scavenging tests (inhibitory concentration [IC50] = 0.89 {\textpm} 0.01 μg/mL, IC50 = 4.20 {\textpm} 1.50 μg/mL). Ethanolic extracts (μg/mL) showed low cytotoxicity on human tumor cell lines (CI50 \>20 μg/mL) for PA, PP, GD, MD. Meanwhile, WO presented cytotoxicity on H460, MCF-7, and K562 tumor cell lines. Conclusion: In our findings, WO and MD extracts revealed a high antioxidant and TPC. WO exhibited better cytotoxic effect compared with 5-FU. Hence, these medicinal plants could be effective to prevent cancer disease and oxidative stress.

}, keywords = {Antioxidant activity, Cancer, cytotoxicity, Oxidative stress, Phytochemical}, doi = {10.4103/pr.pr_109_17}, author = {Oscar Herrera-Calderon and Corina Alvarado-Puray and Jorge Luis Arroyo-Acevedo and Juan Pedro Rojas-Armas and Victor Chumpitaz-Cerrate and Ren{\'a}n Ha{\~n}ari-Quispe and Rita Valenzuela-Herrera} } @article {265, title = {Cytotoxicity Studies of the Extracts, Fractions, and Isolated Compound of Pseudocedrela kotschyi on Cervical Cancer (HeLa), Breast Cancer (MCF-7) and Skeletal Muscle Cancer (RD) Cells}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {February 2017}, pages = {46-50}, type = {Original Article}, chapter = {46}, abstract = {

Background: This study determined the cytotoxic effects of root and stem bark extracts, fractions, and isolated compounds derived from Pseudocedrela kotschyi on HeLa, MCF-7, and RD cells. Materials and Methods: The cytotoxic activity was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay against three cell lines (RD, HeLa, and MCF 7) at concentrations ranging from 0.01 to 1000 μg/mL. Isolation of crude saponin was done from the most active ethyl acetate fraction and further purified using vacuum liquid chromatography and preparative thin layer chromatographic techniques. Results: The cytotoxicity assay revealed that the methanol extract from the root bark and the ethyl acetate fraction from the stem bark exhibited marked anticancer activity with IC50 of 87.36 μg/ml and 21.53 μg/ml, respectively, on HeLa cancer cell line and 101.51 μg/mL and 38.46 μg/mL, respectively, on RD cell line. These values are comparable with that obtained from vinblastine and methotrexate used as standard drugs (IC50 values of 0.01 μg/mL and 0.05 μg/mL, respectively). The isolated crude saponins also gave IC50 values of 5.28 μg/mL and 81.52 μg/mL against the RD cell lines and IC50 values of 1.05 μg/mL and 86.8 μg/mL for the MCF 7 cancer cell lines. PTLC led to the isolation of a compound from the crude saponin which was identified as 7-deacetoxy-7-oxogedunin through spectroscopic analysis and comparison with literature data. Conclusions: P. kotschyi could be considered as a potential source of chemotherapeutic agent. However, further research to determine the exact mechanism of action needs to be carried out.

}, keywords = {3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, 7-deacetoxy-7-oxogedunin, cytotoxicity, HeLa, MCF-7, Pseudocedrela kotschyi, RD}, doi = {10.4103/0974-8490.199776}, author = {Taiwo O Elufioye and Abolaji A Abdul and Jone O Moody} } @article {326, title = {Effects of Ligusticum porteri (Osha) Root Extract on Human Promyelocytic Leukemia Cells}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {April 2017}, pages = {156-160}, type = {Original Article}, chapter = {156}, abstract = {

Background: Ligusticum porteri roots have been traditionally used in folk medicine, but the scientific basis is unclear. Objective: To investigate the cytotoxicity, antioxidant, and immunomodulatory effects of L. porteri root extract on human promyelocytic leukemia (HL-60) cells and H2O2-induced oxidative damaged HL-60 cells. Materials and Methods: HL-60 cells were incubated with different concentrations of root extract, and cells were harvested for viability assays on day 3 and 7. Cytokine levels (interferon-gamma [IFN-γ], interleukin-2 [IL-2], and interleukin-10 [IL-10]) and antioxidant indexes (malondialdehyde [MDA], reduced glutathione [GSH], superoxide dismutase [SOD], and catalase [CAT]) in H2O2-induced-stressed HL-60 were measured after 2 days. Results: The viability of HL-60 challenged with H2O2 declined by 42\% compared to unstressed cells. After 7 days of incubation with 200 or 400 μg/mL L. porteri, the viability of HL-60 cells was two-fold higher than the control. Stressed HL-60 cells treated with 100, 200, and 400 μg/mL L. porteri reduced the lipid peroxidation by 12\%{\textendash}13\%. We noted an increase in GSH levels, SOD and CAT activities in stressed HL-60 supplemented with 400 μg/mL root extract. Treatment with 400 μg/mL L. porteri significantly (P \< 0.05) increased IFN-γ and IL-2 in H2O2-challenged cells. Conclusion: Our data do not support the use of the extract as an antiproliferation and differentiation therapy for acute promyelocytic leukemia. The protective function of L. porteri root extract against oxidative stress could occur through increasing GSH and higher expression of antioxidant enzymes.

}, keywords = {Antioxidative, cytotoxicity, Human promyelocytic leukemia cells, Immune-modulatory, Ligusticum porteri}, doi = {10.4103/0974-8490.204641}, author = {Khanh Nguyen and Jean Sparks and Felix Omoruyi} } @article {256, title = {The Influence of Pluronic F68 and F127 Nanocarrier on Physicochemical Properties, In vitro Release, and Antiproliferative Activity of Thymoquinone Drug}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {February 2017}, pages = {12-20}, type = {Original Article}, chapter = {12}, abstract = {

Background: This study reports on hydrophobic drug thymoquinone (TQ), an active compound found in the volatile oil of Nigella sativa that exhibits anticancer activities. Nanoformulation of this drug could potentially increase its bioavailability to specific target cells. Objective: The aim of this study was to formulate TQ into polymer micelle, Pluronic F127 (5.0 wt \%) and Pluronic F68 (0.1 wt \%), as a drug carrier to enhance its solubility and instability in aqueous media. Materials and Methods: Polymeric micelles encapsulated TQ were prepared by the microwave-assisted solvent evaporation technique. Fourier transform infrared spectroscopy and ultraviolet-visible spectrophotometer were utilized for qualitative confirmation of micelles encapsulation. The surface morphology and mean particle size of the prepared micelles were determined by using transmission electron microscopy (TEM). Cytotoxicity effect was studied using 3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Results: Dynamic laser light scattering (DLS) technique showed hydrodynamic size distribution of optimized micelles of 50 nm, which was in close agreement with the mean particle size obtained from TEM of about 51 nm. Drug release study showed the maximum percentage of TQ release at 61\% after 72 h, while the entrapment efficiency of TQ obtained was 46\% using PF127. The cytotoxic effect of PF127-encapsulated TQ was considerably higher compared to PF68-encapsulated TQ against MCF7 cells, as they exhibited IC50 value of 8 μM and 18 μM, respectively. Conclusion: This study suggests higher molecular weight Pluronic polymer micelles (F127) with hydrophilic-hydrophobic segments which could be used as a suitable candidate for sustainable delivery of TQ. However, comprehensive studies should be carried out to establish the suitability of Pluronic F127 as a carrier for other drugs with similar challenges as TQ.

}, keywords = {cytotoxicity, MCF-7 cell line, Pluronic F68, Thymoquinone-nanoparticle}, doi = {10.4103/0974-8490.199774}, author = {Salwa Shaarani and Shahrul Sahul Hamid and Noor Haida Mohd Kaus} } @article {485, title = {Assessment of Anti-Influenza Activity and Hemagglutination Inhibition of Plumbago indica and Allium sativum Extracts}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {February 2016}, pages = {105-111}, type = {Original Article}, chapter = {105}, abstract = {

Background: Human influenza is a seasonal disease associated with significant morbidity and mortality. Anti-flu ayurvedic/herbal medicines have played a significant role in fighting the virus pandemic. Plumbagin and allicin are commonly used ingredients in many therapeutic remedies, either alone or in conjunction with other natural substances. Evidence suggests that these extracts are associated with a variety of pharmacological activities. Objective: To evaluate anti-influenza activity from Plumbago indica and Allium sativum extract against Influenza A (H1N1)pdm09. Materials and Methods: Different extraction procedures were used to isolate the active ingredient in the solvent system, and quantitative HPLTC confirms the presence of plumbagin and allicin. The cytotoxicity was carried out on Madin-Darby Canine kidney cells, and the 50\% cytotoxic concentration (CC50) values were below 20 mg/mL for both plant extracts. To assess the anti-influenza activity, two assays were employed, simultaneous and posttreatment assay. Results: A. sativum methanolic and ethanolic extracts showed only 14\% reduction in hemagglutination in contrast to P. indica which exhibited 100\% reduction in both simultaneous and posttreatment assay at concentrations of 10 mg/mL, 5 mg/mL, and 1 mg/mL. Conclusions: Our results suggest that P. indica extracts are good candidates for anti-influenza therapy and should be used in medical treatment after further research.

}, keywords = {Allium sativum, Anti-influenza activity, cytotoxicity, Hemagglutination, Plumbago indica}, doi = {10.4103/0974-8490.172562}, author = {Rahul Dilip Chavan and Pramod Shinde and Kaustubh Girkar and Rajendra Madage and Abhay Chowdhary} } @article {515, title = {Evaluation of In vitro Antiviral Activity of Datura metel Linn. Against Rabies Virus}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {August 2016}, pages = {265-269}, type = {Original Article}, chapter = {265}, abstract = {

Objective: The soxhlet and cold extracts of Datura metel Linn. were evaluated for in vitro antirabies activity. Materials and Methods: Soxhlet and cold extraction method were used to extract Datura (fruit and seed) extracts. In vitro cytotoxicity assay was performed by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. Based on the CC50 range, the in vitro antirabies activity of the extracts was screened by rapid fluorescent focus inhibition test and molecular method. Results: The Datura (fruit and seed) extracts were not cytotoxic below 5 mg/ml (CC50). Titer of 10-4 rabies virus challenge virus standard (RV CVS) (1 50\% tissue culture infective dose [1 TCID50]) was obtained by RFFT method and the challenge dose of 10 TCID50 was used for antirabies assay. Datura fruit and seed (soxhlet and cold) extracts showed 50\% inhibition of RV CVS at 2.5 mg/ml and 1.25 mg/ml (inhibitory concentration 50\% [IC50]), respectively. The tested extracts showed selectivity index (CC50/IC50) ranging from 2 to 4. The viral RNA was extracted and real-time reverse transcription-polymerase chain reaction was performed which also revealed a 2-fold reduction of viral load at 1.25 mg/ml of the Datura seed (soxhlet methanolic and cold aqueous) extracts. Conclusion: To the best of our knowledge, this is the first study of in vitro antiviral activity of D. metel Linn. against rabies virus. Datura seed extracts have a potential in vitro antirabies activity and, in future, can be further screened for in vivo activity against rabies virus in murine model.

}, keywords = {Antirabies, Antiviral assay, cytotoxicity, Datura, Rapid fluorescent focus inhibition test}, doi = {10.4103/0974-8490.188874}, author = {Soumen Roy and Sandeepan Mukherjee and Sandip Pawar and Abhay Chowdhary} } @article {488, title = {Evaluation of In Vitro Cytotoxic and Antioxidant Activity of Datura metel Linn. and Cynodon dactylon Linn. Extracts}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {February 2016}, pages = {123-127}, type = {Original Article}, chapter = {123}, abstract = {

Aim: To evaluate in vitro cytotoxicity and antioxidant activity of Datura metel L. and Cynodon dactylon L. extracts. Materials and Methods: The extraction of plants parts (datura seed and fruit pulp) and areal parts of durva was carried out using soxhlet and cold extraction method using solvents namely methanol and distilled water. The total phenolic content (TPC) and total flavonoid content (TFC) was determined by established methods. The in vitro cytotoxicity assay was performed in vero cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay method. In vitro antioxidant activity of the extract was performed by 2, 2-diphenyl-1-picrylhydrazyl radical scavenging method. Results: We found that the highest amount of TPC and TFC in methanolic extracts of seed (268.6 μg of gallic acid equivalence/mg of dry plant material) and fruit pulp (8.84 μg of quercetin equivalence/mg dry plant material) of D. metel, respectively prepared by Soxhlet method. The methanolic extract of C. dactylon prepared using soxhlation has shown potent free radical scavenging activity with 50\% inhibitory concentration (IC50) value of 100 μg/ ml. The IC50 of a methanolic cold extract of datura fruit was found to be 3 mg/ml against vero cell line. Conclusion: We observed that plant parts of C. dactylon and D. metel have a high antioxidant activity. Further research is needed to explore the therapeutic potential of these plant extracts.

}, keywords = {2, 2-diphenyl-1-picrylhydrazyl radical, Antioxidant, cytotoxicity, Total flavonoid content, Total phenolic content}, doi = {10.4103/0974-8490.175610}, author = {Soumen Roy and Sandip Pawar and Abhay Chowdhary} } @article {509, title = {Probing Regenerative Potential of Moringa oleifera Aqueous Extracts Using In vitro Cellular Assays}, journal = {Pharmacognosy Research}, volume = {8}, year = {2016}, month = {August 2016}, pages = {231-237}, type = {Original Article}, chapter = {231}, abstract = {

Background: Molecules stimulating regeneration and proliferation of cells are of significance in combating ailments caused due to tissue injury, inflammation, and degenerative disorders. Moringa oleifera is one of the most valued food plants having the profile of important nutrients and impressive range of medicinal uses. Objective: To evaluate the potential of M. oleifera aqueous leaf and flower extracts to promote the proliferation of cells and explore their effect on cancer cell lines for assessment of safety. Materials and Methods: Aqueous leaf and flower extracts of M. oleifera were investigated for effect on rat-derived primary fibroblast, mesenchymal stem cells (MSCs), and cancer cell lines using cell proliferation assay. They were also tested and compared for wound healing, angiogenesis, and hepatoprotective effect using in vitro assays. Results: Statistically significant increase in the proliferation of primary rat fibroblast, MSCs, and angiogenesis was observed after treatment with aqueous flower extract. The aqueous leaf extract determined a comparatively moderate increment in the proliferation of MSCs and angiogenesis. It however showed prominent cytotoxicity to cancer cell lines and a significant hepatoprotective effect. Conclusion: A very clear difference in response of the two extracts to different types of cells was detected in this study. The aqueous flower extract exhibited a higher potential to stimulate cell proliferation while not exerting the same effect on cancer cell lines. The leaf extract on the other hand, had a prominent antitumor and hepatoptotective effects.

}, keywords = {Aqueous, cytotoxicity, Fibroblast, in vitro, Viability}, doi = {10.4103/0974-8490.188877}, author = {Evangeline E. Fernandes and Anubha V. Pulwale and Gauri A. Patil and Alpana S. Moghe} } @article {386, title = {Cytotoxic Activity of ten Aagae from the Persian Gulf and Oman Sea on Human Beast Cancer cell lines; MDA-MB-231, MCF-7, and T-47D}, journal = {Pharmacognosy Research}, volume = {7}, year = {2015}, month = {June 2015}, pages = {133-137}, type = {Original Article }, chapter = {133}, abstract = {

Background:\ Seaweeds have proven to be a promising natural source of bioactive metabolites for drug development.\ Objective:\ This study aimed to monitor the ethanol extract of ten algae from the Persian Gulf and Oman Sea, for their\ in vitro\ cytotoxic activity on three human breast cancer cell lines.\ MaterialsandMethods:\ Three human breast cancer cell lines including MDA-MB-231(ER-\ ), MCF-7(ER\ +\ ), and T-47D (ER\ +\ ) were treated by different concentrations of total ethanol (90\%) algae extracts and the cytotoxic effects were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Doxorubicin (Ebewe, Austria) was used as a positive control. After 72 h of incubation, the cytotoxic effect of the algae was calculated and presented as 50\%-inhibitory concentration (IC50\ ).\ Results:\ The results indicated\ Gracilariafoliifera\ and\ Cladophoropsis\ sp. to be the most active algae in terms of cytotoxic effects on the investigated cancer cell lines. The IC\ 50\ values against MDA-MB-231, MCF-7, and T-47D cells were, respectively, 74.89 ΁ 21.71, 207.81 ΁ 12.07, and 203.25 ΁ 30.98 mg/ml for\ G.foliifera\ and 66.48 ΁ 4.96, 150.86 ΁ 51.56 and \>400 mg/ml for\ Cladophoropsis\ sp. The rest of the algal extracts were observed not to have significant cytotoxic effects in the concentration range from 6.25 mg/ml to 400 mg/ml.\ Conclusion:\ Our data conclusively suggest that\ G.foliifera\ and\ Cladophoropsis\ sp. may be good candidates for further fractionation to obtain novel anticancer substances. Moreover, stronger cytotoxic effects on estrogen negative breast cancer cell line (MDA-MB-231(ER-\ )) in comparison to estrogen positive cells (MCF-7 and T-47D) suggest that the extract of\ G.foliifera\ and\ Cladophoropsis\ sp. may have an estrogen receptor/progesterone receptor-independent mechanism for their cellular growth inhibition.

}, keywords = {3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, Algae, cytotoxicity, Oman Sea, Persian Gulf}, doi = {10.4103/0974-8490.150539 }, author = {Nasrollah Erfani and Zahra Nazemosadat and Mahmoodreza Moein} } @article {466, title = {Evaluation of antiviral activity of essential oil of Trachyspermum Ammi against Japanese encephalitis virus}, journal = {Pharmacognosy Research}, volume = {7}, year = {2015}, month = {June,2015}, pages = {263-267}, type = {Original Article}, chapter = {263}, abstract = {

Background: Japanese encephalitis is a leading form of viral encephalitis, prevalent mostly in South Eastern Asia caused by Japanese encephalitis virus (JEV). It is transmitted by the mosquitoes of the Culex sp. The disease affects children and results in 50\% result in permanent neuropsychiatric disorder. There arises a need to develop a safe, affordable, and potent anti-viral agent against JEV. This study aimed to assess the antiviral activity of ajwain (Trachyspermum ammi: Umbellifereae) essential oil against JEV. Materials and Methods: Ajwain oil was extracted by distillation method and in vitro cytotoxicity assay was performed in vero cell line by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay method. JEV titer was determined by plaque assay and in vitro antiviral activity of ajwain oil was quantified by the plaque reduction neutralization test (PRNT). Results: Cytotoxic concentration of the oil was found to be 1 mg/ml by MTT assay. The titer of the virus pool was found to be 50{\texttimes} 107 PFU/ml. we observed 80\% and 40\% virus inhibition in 0.5mg/ml of ajwain oil by PRNT method in preexposure treatment and postexposure treatment (antiviral activity), respectively. Conclusion: Our data indicate ajwain oil has potential in vitro antiviral activity against JEV. Further, the active biomolecule will be purified and evaluated for anti-JEV activity and also to scale up for in vivo trial to evaluate the efficacy of ajwain oil in future.

}, keywords = {Ajwain oil, Antiviral, cytotoxicity, Japanese encephalitis virus.}, doi = {10.4103/0974-8490.157977}, author = {Soumen Roy and Pratibha Chaurvedi and Abhay Chowdhary} } @article {856, title = {Anticancer Potential of Syzygium aromaticum L. in MCF-7 Human Breast Cancer Cell Lines}, journal = {Pharmacognosy Research}, volume = {6}, year = {2014}, month = {August,2014}, pages = {350-354}, type = {Short Communication}, chapter = {350}, abstract = {

Background: The common treatment for cancer is unfavorable because it causes many detrimental side effects, and lately, there has been a growing resistance toward anticancer drugs, which worsens the future of cancer treatment. Therefore, the focus has now shifted toward natural products, such as spices and plants, among many others, to save the future of cancer treatment. Cloves (Syzygium aromaticum L.) are spices with the highest antioxidant content among natural products. Besides acting as an antioxidant, cloves also possess many other functions, such as anti-inflammatory, antibacterial, and antiseptic, which makes them an ideal natural source to be developed as an anticancer agent. Objective: This study aims to evaluate the cytotoxic activity of cloves toward MCF-7 human breast cancer cell lines. Materials and Methods: Different concentrations of water extract, ethanol extract, and essential oil of cloves were investigated for their anticancer potential in vitro through a brine shrimp lethality test (BSLT) and an MTT assay. Results: In both BSLT and MTT assays, the essential oil showed the highest cytotoxic effect, followed by ethanol and water extract. The LD 50 concentration of essential oil in the 24 hr\ BSLT was 37 μg/mL. Furthermore, the IC 50 values in the 24 hr\ and 48 hr\ MTT assays of the essential oil were 36.43 μg/mL and 17.6 μg/mL, respectively. Conclusion: Cloves are natural products with excellent cytotoxicity toward MCF-7 cells; thus, they are promising sources for the development of anticancer agents.

}, keywords = {Antioxidant, Brine shrimp lethality test, Clove, cytotoxicity, MCF-7}, doi = {10.4103/0974-8490.138291}, author = {Parvinnesh S Kumar and Raden M Febriyanti and Ferry F Sofyan and Dimas E Luftimas and Rizky Abdulah} } @article {665, title = {Cellular Responses with Thymoquinone Treatment in Human Breast Cancer cell line MCF-7}, journal = {Pharmacognosy Research}, volume = {5}, year = {2013}, month = {May 2013}, pages = {200-206}, type = {Original Article }, chapter = {200}, abstract = {

Background:Nigella sativa\ or black seed extract has been reported to show various medicinal benefits. Thymoquinone which is an active compound of its seed has been reported to contain anti-cancer properties.\ Objective:\ The study addressed the anti-cancer efficiency of long-term\ in vitro\ treatment with thymoquinone towards human breast cancer cell lines MCF-7.\ Materials and Methods:\ Cell proliferation was determined with CellTiter 96 Aqueous. Non-Radioactive Cell Proliferation Assay Kit. It was followed with trypan blue exclusion test to determine the percentage of viable cells. The study incorporated cell cycle assay to distinguish cell distribution at various cell cycle phases using Cycletest Plus DNA Reagent Kit. The apoptosis detection kit was used to determine the percentage of apoptotic and necrotic cells using flow cytometry.\ Results:\ The 50\% inhibitory concentration (IC\ 50\ ) value determined using the proliferation assay was 25 μM thymoquinone. Late apoptotic cell percentage increased rapidly when treatment duration was increased to 24 h with 25 and 100 μM thymoquinone. Further analysis using cell cycle assay showed thymoquinone inhibition of breast cancer cell proliferation at minimal dose 25 μM and led to S phase arrest significantly at 72 h treatment (P\ = 0.009). It was also noted elevation sub-G\ 1\ peak following treatment with 25 μM thymoquinone for 12 h. Increase in thymoquinone to 50 μM caused G\ 2\ phase arrest at each time-point studied.\ Conclusion:\ In general thymoquinone showed sustained inhibition of breast cancer cell proliferation with long-term treatment. Specificity of phase arrest was determined by thymoquinone dose.

}, keywords = {Apoptosis, cytotoxicity, Long-term Exposure, MCF-7 cell line, Thymoquinone}, doi = {10.4103/0974-8490.112428}, author = {Marjaneh Motaghed and Faisal Muti Al-Hassan and Shahrul Sahul Hamid} } @article {618, title = {Volatile Constituents of Pinus Roxburghii from Nepal}, journal = {Pharmacognosy Research}, volume = {5}, year = {2013}, month = {January 2013}, pages = {43-48}, type = { Original Article}, chapter = {43}, abstract = {

Background:Pinus roxburghii\ Sarg. Is one of 3 species of pine found in Nepal, the oil of which is traditionally used to treat cuts, wounds, boils, and blisters.\ Objective:\ To obtain, analyze, and examine the anti-microbial and cytotoxic activities of the essential oils of\ P. roxburghii.\ Materials and Methods:\ Three plant parts (cone, needle, and bark) of\ Pinus roxburghii\ were collected in Biratnagar, Nepal. The essential oils were obtained by hydrodistillation, and the chemical compositions were determined by GC-MS. The needle and cone essential oils were screened for anti-microbial activity against\ Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa,Escherichia coli, and\ Aspergillus niger;\ brine shrimp (Artemia salina) lethality; and\ in-vitro\ cytotoxicity against MCF-7 cells.\ Results:\ GC-MS analysis for the cone oil revealed 81 compounds with 78 components being identified (95.5\% of the oil) while 98.3\% of needle oil was identified to contain 68 components and 98.6\% of the bark oil (38 components) was identified. The 3 essential oils were dominated by sesquiterpenes, particularly (E)-caryophyllene (26.8\%-34.5\%) and α-humulene (5.0\%-7.3\%) as well as monoterpene alcohols terpinen-4-ol (4.1\%-30.1\%) and α-terpineol(2.8\%-5.0\%). The monoterpene δ-3-carene was present only in needle and cone essential oils (2.3\% and 6.8\%, respectively). Bio-activity assays of the cone essential oil of\ P. roxburghii\ showed remarkable cytotoxic activity (100\% killing of MCF-7 cells at 100 μg/mL) along with notable brine shrimp lethality (LC50\ =11.8 μg/mL). The cone essential oil did not show anti-bacterial activity, but it did exhibit anti-fungal activity against\ Aspergillus niger\ (MIC=39 μg/mL).\ Conclusion:\ The bioactivity of\ P. roxburghii\ essential oil is consistent with its traditional medicinal use.

}, keywords = {Anti-fungal, Brine shrimp lethality, cytotoxicity, Essential oil composition, α-humulene}, doi = {10.4103/0974-8490.105650}, author = {Prabodh Satyal and Prajwal Paudel and Josna Raut and Akash Deo and Noura S Dosoky and William N Setzer} } @article {902, title = {Antioxidant, cytotoxic, antitumor, and protective DNA damage metabolites from the red sea brown alga Sargassum sp}, journal = {Pharmacognosy Research}, volume = {3}, year = {2011}, month = {September 2011}, pages = {160-165}, type = {Original Article}, chapter = {160}, abstract = {

Background: Macroalgae can be viewed as a potential antioxidant and anti-inflammatory sources owing to their capability of producing compounds for its protection from environmental factors such as heat, pollution, stress, oxygen concentration, and UV radiations. Objective: To isolate major compounds which are mainly responsible for the pharmacological activity of brown alga under investigation, Sargassum sp. Materials and Methods: Algal material was air dried, extracted with a mixture of organic solvents, and fractionated with different adsorbents. The structures of obtained pure compounds were elucidated with different spectroscopic techniques, and two pure materials were tested for protection of DNA from damage, antioxidant, antitumor, and cytotoxicity. Results: Four pure compounds were obtained, of which fucosterol (1) and fucoxanthin (4) were tested; it was found that fucoxanthin has strong antioxidant and cytotoxicity against breast cancer (MCF-7) with IC 50 = 11.5 ΅g/ml. Conclusion: The naturally highly conjugated safe compound fucoxanthin could be used as antioxidant and as an antitumor compound.

}, keywords = {Antioxidant, cytotoxicity, Fucoxanthin, Sargassum sp}, doi = {10.4103/0974-8490.85000}, author = {Seif-Eldin N Ayyad and Saleh T Ezmirly and Salim A Basaif and Walied M Alarif and Adel F Badria and Farid A Badria} } @article {937, title = {A Comparison of the Cytotoxic Potential of Standardized Aqueous and Ethanolic Extracts of a Polyherbal Mixture Comprised of Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizome)}, journal = {Pharmacognosy Research}, volume = {2}, year = {2010}, month = {January 2011}, pages = {335-342}, type = {Original Article}, chapter = {335}, abstract = {

Background: A decoction (hot-water extract) comprised of Nigella sativa (seeds), Hemidesmus indicus (roots), and Smilax glabra (rhizome) has been reported to prevent chemically-induced hepatocarcinogenic changes in rats and to exert significant cytotoxic effects on human hepatoma (HepG2) cells. However, the decoction used in previous studies to determine cytotoxicity was not standardized. Further, during preparation of pharmaceuticals for clinical use, it is more convenient to use an ethanolic extract. Therefore this study was carried out to (a) develop standardized aqueous and ethanolic extracts of the plant mixture (N. sativa, H. indicus, and S. glabra) used in the preparation of the original decoction, and (b) compare the cytotoxic effects of these two extracts by evaluating cytotoxicity to the human hepatoma (HepG2) cell line. Methods: Aqueous and ethanolic extracts have been standardized by evaluating organoleptic characters, physicochemical properties, qualitative and quantitative analysis of chemical constituents, and analysis of High Performance Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) profiles. Cytotoxic potentials of the above standardized extracts were compared by evaluating their effects on the survival and overall cell activity of HepG2 cells by use of the 3-(4, 5-dimethylthiazol-2yl) -2, 5 - biphenyl tetrazolium bromide (MTT) and Sulphorhodamine B (SRB) assays. Results: Results from MTT and SRB assays demonstrated that both extracts exerted strong dose-dependent in vitro cytotoxicity to HepG2 cells. The standardized aqueous extract showed a marginally (though significantly, P\<0.05) higher cyotoxic potential than the ethanolic extract. Thymoquinone, an already known cytotoxic compound isolated from N. sativa seeds was only observed in the standardized ethanolic extract. Thus, compounds other than thymoquinone appear to mediate the cytotoxicity of the standardized aqueous extract of this poly-herbal preparation. Conclusion: It may be concluded that results obtained in the present study could be used as a diagnostic tool for the correct identification of these aqueous or ethanolic extracts and would be useful for the preparation of a standardized pharmaceutical product that may be used in the future for clinical therapy of hepatocellular carcinoma

}, keywords = {cytotoxicity, Hemidesmus indicus, MTT and SRB assays, Nigella sativa, Smilax glabra, Standardization}, doi = {10.4103/0974-8490.75451}, author = {Sameera R Samarakoon and Ira Thabrew and Prasanna B Galhena and Dilip De Silva and Kamani H Tennekoon} } @article {877, title = {Analysis of Total Proteins, Polysaccharides and Glycosaponins Contents of Orthosiphon stamineus Benth. In Spray and Freeze Dried Methanol: Water(1:1) extract and its Contribution to Cytotoxic and Antiangiogenic Activities}, journal = {Pharmacognosy Research}, volume = {1}, year = {2009}, month = {January 2010 }, pages = {302-326}, type = {Original Article}, chapter = {320}, abstract = {

Orthosiphon stamineus (OS), Benth, (Lamiaceae), popularly known as Java tea, is a useful traditional medicinal herb cultivated largely in Malaysia and in Indonesia. This study was conducted to investigate the primary metabolic contents, cytotoxicity and antiangiogenic activity of freeze dried and spray dried extracts of the OS methanol: water (1:1). Primary metabolic contents were evaluated for total proteins, total polysaccharides and glycosaponins (P\<0.05). Freeze dried extract showed high content of proteins and glycosaponins while contents of polysaccharides were high in spray dried extracts. Cytotoxicity was detected by MTT assay using MDA-MB-231 breast cancer cell line and anti angiogenesis assay was performed on rat aorta. Both extract showed moderate cytotoxic activity using extract in high and medium concentration. Freeze dried extract showed strong antiangiogenic activity while spray dried extract showed no activity. A significant change in total proteins of freeze dried extract may be the reason for strong antiangiogenic activity.

}, keywords = {Antiangiogenesis, cytotoxicity, Glycosaponins, Orthosiphon stamineus, Polysaccharides, Proteins}, doi = {Nil}, author = {Siddiqui M.J and Hafizoh S. N and Ismail Z. and Sahib H.B and Helal M.H.S and Majid A.M.S Abdul} } @article {729, title = {Effects of Ephedrine and Ephedra fragilis Crude Extracts on Human Peripheral Lymphocytes}, journal = {Pharmacognosy Research}, volume = {1}, year = {2009}, month = {January 2010 }, pages = {38-42}, type = {Original Article}, chapter = {38}, abstract = {

Ephedra fragilis Desf. (Ephedraceae), a locally cultivated medicinal plant, is a source of ephedrine and pseudoephedrine, two important alkaloids, which have long played an important pharmacological role. The present study investigated the in vitro effects of ephedrine and the Ephedra branch extract on unstimulated lymphocytes. Ephedra alkaloids were extracted from various plant parts and after phytochemical analysis, the brine shrimp lethality test was used to determine the activity of the extracts. The LC50 of the branch extract was 581.395 μg/ml, showing no statistical difference from that of ephedrine (208.203 μg/ml). The ephedrine and extract did not show toxic effects on lymphocytes but exhibited immunostimulant activity. Although ephedrine cannot be used as an immune booster, it can be used as a lead drug for further immunological research.

}, keywords = {cell proliferation, cytotoxicity, Ephedra fragilis, Ephedrine, Lymphocyte activation}, doi = {Nil}, author = {E. Attard and K. Vella} }