Mitochondria-Mediated Moderation of Apoptosis by EGCG in Cytotoxic Neuronal Cells Induced by Lead (Pb) and Amyloid Peptides.

The developmental, epigenetic, and epidemiological studies on lead (Pb) toxicity have reported a strong connection between lead exposure and the progression of Alzheimer's disease (AD). The amyloid peptides were the main triggering elements, in the generation of extracellular plaques through which multiple cellular signaling events such as apoptosis and primarily oxidant-antioxidant balancing system will be affected, which leads to neuronal cell death. Our previous studies indicated that epigallocatechin gallate (EGCG), abundantly present in green tea, was found to be effective in alleviating the metal-induced neurotoxicity at the cellular level in terms of cell viability and apoptosis The aim of this study was to explore the protective mechanism of EGCG on the markers of oxidant-antioxidant system and mitochondria, which are involved in metal-induced neuronal cell death. Initially, the IC50 values for lead(Pb-5 µM), amyloid peptides (AP(1-40)-60 µM; AP(1-40)-8 µM), and EC50 value for EGCG(50 µM) were determined by both time- (12 h, 24 h, 48 h) and concentration-dependent manner and analyzed by MTT assay. The experimental groups were designed initially by treating with Pb and APs individually and in different combinations along with the presence of EGCG and are compared to the Pb and AP treated group without EGCG exposure. The cell lysates were used for analyzing oxidative stress markers by standardized laboratory protocol and the expression of mitochondrial markers such as VDAC and cytochrome C which were analyzed by both western blot and real-time PCR. Our results indicate that the EGCG-treated group has shown a significant increase in antioxidant marker expression levels (GSH, SOD, catalase, vitamin C) and a decrease in oxidative stress marker (NOS, MDA) levels when compared to the group without EGCG treatment (p < 0.05). Similarly, a significant decrease in expression levels of VDAC and cytochrome c were observed in the EGCG-treated group when compared to the group without EGCG treatment (p < 0.05). Our approach revealed that EGCG protects SH-SY5Y cells from Pb- and AP-induced cytotoxicity by regulating voltage-dependent anion channel (VDAC) expression and oxidant-antioxidant system and inhibits neuronal cell death.