Potent in vitro Antioxidant, Antimicrobial, and Antidiabetic Effects of a Polyherbal Mixture: Evidence from 3T3-L1, β-TC6, and C2C12 Cell Line Studies

Aim and Objectives: The traditional plants Plumeria pudica, Podranea ricasoliana, and Leea macrophylla have been used for their medicinal properties since ancient times. Our main objective is to identify the preliminary chemicals of these plant leaves to prepare a polyherbal mixture and apply it for in vitro DPPH, H₂O₂, and NO scavenging free radical antioxidants test with antibacterial study and evaluated for its in vitro antidiabetic potential using 3T3-L1, β-TC6, and C2C12 cell lines. Materials and Methods: The macroscopic study as well as the preliminary phytochemical screening of these plants, possess many numbers of metabolites that are responsible for anti-inflammatory and anti-oxidant properties. The plant extract's hydrogen peroxide scavenging activity was assessed by measuring absorbance at 230 nm using a UV-vis spectrophotometer. The disc plate technique uses antibiotics to decrease the various gram-positive and negative bacterial properties. Ethanol-based extract of Polyherbal Mixture (PHM) was subjected to phytochemical analysis. The increase of glucose uptake in 3T3L1 cells (a) and C2C12 myoblast cells over the treatment of PHM extract at varying concentrations ranging from 10 to 500 μg/mL. Metformin and Insulin-stimulated glucose uptake were considered positive controls. Experiments were conducted in triplicate. Results: The IC₅₀ values of the DPPH, NO, and H2O2 scavenging assays revealed that the test compounds were less potent than the standard drugs, yet they still exhibited satisfactory free radical scavenging activity. The ethanolic extract of the Polyherbal Mixture (PHM) showed antibacterial activity that was slightly less potent than the standard antibiotic ciprofloxacin, yet still produced satisfactory results with comparable inhibition zones. Furthermore, the IC₅₀ values of the test drug PHM (16.71, 12.0, and 20.8 μg/mL) and the standard drug ascorbic acid (7.07, 6.07, and 8.0 μg/mL) indicate that PHM possesses antioxidant scavenging activity. The zone of inhibition of PHM was satisfactory as compared to the standard drug during the antimicrobial study. Specifically, in 3T3-L1 cells, PHM-500 (2.9±0.25), metformin (2.8±0.41), and insulin (5.9±0.43) increased glucose uptake compared to control. Similarly, in C2C12 myoblasts, PHM-500 (4.37±0.28), metformin (4.91±0.34), and insulin (6.77±0.32) showed increased glucose uptake. Conclusion: The poly-herbal mixture is expected to exhibit beneficial anti-microbial, anti-oxidant, and anti-diabetic activities, making it a unique and unpublished formulation.