Phytochemical analysis and in vitro cytostatic potential of ethnopharmacological important medicinal plants

Toxicol Rep. 2020 Feb 25:7:443-452. doi: 10.1016/j.toxrep.2020.02.016. eCollection 2020.

Abstract

Annonareticulate (Mart.), Lablab purpureus (L.) Sweet, Murrayakoenigii (L.) Spreng, Moringaoleifera (Lam.), Hibiscussabdariffa (L.) and Euphorbiahirta (L.) are most commonly used medicinal plants by the traditional healers of Karbi Anglong district of Assam, India against different human ailments including cancer suspected cases. The proposed study includes field survey related to ethnomedicinal aspects of medicinal plants, phytochemical analysis, and evaluation of their cytostatic potential with the possible mode of action against Dalton's lymphoma (DL) cell line. The phytochemical analysis of all the plant's extract was studied using standard protocol. The cytotoxicity of the methanol extracts was determined by MTT reduction assay. The effect of the same extract was also tested for development of apoptosis features in DL cells using a fluorescence microscope and flowcytometry. The underlying mechanism closely associated with apoptotic cell death was also studied by measuring reactive oxygen species (ROS), mitochondrial membrane potential, and expression level of apoptosis inducing proteins. Murraya koenigii induced more apoptotic features in DL cells, followed by Annona reticulate. The decrease in mitochondrial membrane potential, release of cytochrome- c, increase in ROS level and higher expression of caspases (3 and 9) after plant extracts treatment may cause involvement of mitochondria in the process of apoptosis. From this study, it can be concluded that the plant species mainly Murraya koenigi and Annona reticulate significantly induced cytotoxicity in DL cells through apoptosis by utilizing mitochondrial pathway.

Keywords: AO/EB, acridine orange and ethidium bromide; Annona reticulate; DL, Dalton’s ascites lymphoma; DPPH, 2,2-diphenyl-1-picrylhydrazyl; Euphorbia hirta; FL, fidelity level; Hibiscus sabdariffa; IU, International unit; Lablab purpureus; MTT, 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide; Moringa oleifera; Murraya koenigii; OD, Optical density; PBMC, peripheral blood mononuclear cell; RFC, Relative frequency of citation; ROS, reactive oxygen species; ΔΨm, mitochondrial membrane potential.