NEURO-PROTECTIVE PROPERTIES OF ORTHOSIPHON STAMINUS (BENTH) LEAF METHANOLIC FRACTION THROUGH ANTIOXIDANT MECHANISMS ON SH-SY5Y CELLS: AN IN-VITRO EVALUATION

In present study to evaluate the mechanisms behind the antioxidant and neuro-protective potential of Orthosiphon staminus methanol bio active guided fraction (OMF) using SH-SY5Y human neuroblastoma cell culture model was carried out. In in-vitro studies, mechanisms of neutralization of H₂O₂ induced toxicity by OMF using MTT, LDH, ROS, MMP, MDA, Comet assay was performed. Regulation of anti-oxidant enzyme levels by western blot analysis and neuronal biomarker genes (BDNF, TH and AADC genes) at genetic level by quantitative RTPCR was performed. Cells pretreated with OMF effectively prevents the H₂O₂ induced anti cell proliferation (cell viability), oxidative stress, genotoxicity and significantly enhance (~ 6 fold, p < 0.01) the expression of antioxidant enzymes and increase in (~ 4 fold, p < 0.01) neuronal biomarker gene expression. Present study demonstrated that, significant neuro-protective activity of O. staminus methanol bioactive guided fraction against H₂O₂ induced oxidative stress by antioxidant mechanisms and enhancement of neuronal biomarker gene expression. This study validate its use by folk medicinal practitioners in India. Therefore, OMF of O. staminus can serve as a strong candidate for development of herbal neuro-protective compositions.