@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 {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 {337, title = {Attenuation of Methotrexate-induced Hepatorenal Damage by Terminalia bellerica Fruit Extract in Experimental Rats}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {July 2018}, pages = {301-308}, type = {Original Article}, chapter = {301}, abstract = {

Background: Methotrexate (MTX) is used for numerous malignancies and autoimmune disorders. With such widespread use, MTX-induced hepatorenal toxicity is an issue of concern that still needs to be addressed. Objective: The aim of the present study is to evaluate the role of Terminalia bellerica extract (TBE) in MTX-induced hepatorenal toxicity in Wistar albino rats. Materials and Methods: Rats were randomly divided into six groups (n = 6) {\textendash} received MTX 20 mg/kg intraperitoneally on the 4th day along with pretreatment with different doses of TBE (100 mg/kg, 200 mg/kg, and 400 mg/kg, p.o) given from 1st to 15th day. MTX-induced hepatorenal toxicity was evaluated by biochemical hepatic and renal parameters along with histopathology and immunohistochemistry. Results: Hepatorenal toxicity induced by MTX was attributed to increased oxidative stress, biochemical liver, and kidney parameters and upregulation of caspase-3 and nuclear factor kappa B (NFkB). MTX-treated group observed twofold to threefold rise in aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine values{\textendash}138.49 IU/L, 125.81 IU/L, 63.09 mg/dl, and 1.895 mg/dl, respectively. Groups pretreated with TBE (400 mg/kg) observed a significant decrease (P \< 0.001) in oxidative stress and biochemical parameters {\textendash} AST (63.94 IU/L), ALT (55.98 IU/L), BUN (37.02 mg/dl), and creatinine (1.065 mg/dl). Pretreatment with TBE 400 mg/kg, histopathology of both liver and kidney tissues showed improved architectural damage and immunohistochemistry showed downregulation of increased antigens-caspase-3 and NFkB. Conclusion: T. bellerica fruit extract (400 mg/kg) showed significant hepatorenal protection by reducing oxidative stress, elevating serum enzymes, and downregulating the tissue expressions of caspase-3 and NFkB.

}, keywords = {caspase-3, Hepatotoxicity, Methotrexate, Nephrotoxicity, Nuclear factor kappa B, Terminalia bellerica}, doi = {10.4103/pr.pr_159_17}, author = {Prerna Chauhan and Himanshu Sharma and Surender Singh and Yogendra Kumar Gupta and Uma Kumar} } @article {338, title = {Evaluation of AntiCancer Activity of Methanolic Extract of Hiptage benghalensis (L.) Kurz on Cancer Cell Lines}, journal = {Pharmacognosy Research}, volume = {10}, year = {2018}, month = {July 2018}, pages = {309-313}, type = {Original Article}, chapter = {309}, abstract = {

Aim and Background: This study aimed to study the anticancer effects of Hiptage benghalensis (L.) kurz using various human cancer cell cultures, in vitro using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay. Materials and Methods: Human cervical carcinoma (HeLa) cells, human breast cancer (MCF-7) cells, and human neuroblastoma (IMR-32) cells were maintained in a 5\% CO2 incubator at 37{\textdegree}C. Different concentrations of an extract of H. benghalensis in serum-free culture medium were freshly prepared and used for cytotoxic activity using MTT assay activity. Results: Among the plant extract had revealed that, the greater percentage inhibition in all types of cancer cells in a dose-dependent manner using MTT assay. The concentration of the extract causing 50\% cell death values of methanolic extract of H. benghalensis were found to be 50.73, 47.90, and 53.76 μg/mL against HeLa, MCF-7, and IMR-32, respectively. Conclusion: H. benghalensis methanolic extract were showed increased percentage inhibition of MCF 7, HeLa, and IMR 32 cells using MTT assay.

}, keywords = {caspase-3, Cell viability, Hiptage benghalensis, Reactive oxygen species}, doi = {10.4103/pr.pr_102_17}, author = {Babu Rao Bhukya and Narsimha Reddy Yellu} } @article {456, title = {Annonacin Exerts Antitumor Activity through Induction of Apoptosis and Extracellular Signal-regulated Kinase Inhibition}, journal = {Pharmacognosy Research}, volume = {9}, year = {2017}, month = {November 2017}, pages = {378-383}, type = {Original Article}, chapter = {378}, abstract = {

Background: Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries. Annonacin, a natural pure compound extracted from the seeds of Annona muricata, is a potential alternative therapeutic agent to treat EC. Objective: To study the antitumor activity of annonacin and its mechanism of action in EC cells (ECCs). Materials and Methods: Viability of ECCs treated with annonacin for 72 h was determined using methyl thiazolyl tetrazolium assay. The induction of cell cycle arrest and apoptotic cell death was evaluated using propidium iodide and annexin V-PE/7-AAD assay, respectively. DNA strand breaks were visualized using transferase dUTP nick end labeling assay, and the effects of annonacin on survival signaling were determined using western blotting. Results: Annonacin exhibited antiproliferative effects on EC cell lines (ECC-1 and HEC-1A) and primary cells (EC6-ept and EC14-ept) with EC50 values ranging from 4.62 to 4.92 {\textmu}g/ml. EC cells were shown arrested at G2/M phase after treated with 4 {\textmu}g/ml of annonacin for 72 h. This led to a significant increase in apoptotic cell death (65.7\%) in these cells when compared to vehicle-treated cells (P \< 0.005). We further showed that annonacin-mediated apoptotic cell death was associated with an increase in caspase-3 cleavage and DNA fragmentation. Cell apoptosis was accompanied with downregulation of extracellular signal-regulated kinase survival protein expression and induction of G2/M cell cycle arrest. Conclusion: Annonacin may be a potential novel therapeutic agent for EC patients.

}, keywords = {Annona muricata, Apoptosis, caspase-3, Cell cycle arrest, Extracellular signal-regulated kinase, Uterine cancer}, doi = {10.4103/pr.pr_19_17}, author = {Chee Voon Yap and Kavita S Subramaniam and Sik Wey Khor and Ivy Chung} }