Syzigium aromaticum may protect against paraquat mediated toxicity via p53 and Sestrin2 modulation

Abstract

AbstractBackgroundNumerous environmental risk factors, particularly neurotoxins, have been linked to the development of sporadic Parkinson’s disease (PD), even though its exact cause is still unknown. The common spice Syzigium aromaticum has the greatest ORAC (Oxygen Radical Absorbance Capacity) value according to the National Institute on Aging. One of the three components of healthy ageing, vata, is calmed by Syzigium aromaticum while also stimulates nerves. The study investigated the rescue effect of Syzigium aromaticum ethanolic extract against paraquat (PQ) mediated PD type toxicity and elucidated the underlying molecular mechanism using the cell line.MethodThe cold extraction method was used to prepare the Syzigium aromaticum ethanolic extract by powdering, soaking in ethanol and stirring for 72 h. Then the supernatant was gradually aliquoted and filtered with Whatman filter paper no 1. Paraquat, at different doses, was used to generate PD type toxicity in SH‐SY5Y cells for 24, 48, and 72 hours, and MTT test was used to measure the cytotoxicity. The rescue effect was demonstrated by simultaneously treating cells with 100 mM PQ and various doses of Syzigium aromaticum and then performing MTT assay. The protein expression studies was done using western blot technique to elucidate the underlying mechanism.ResultThe toxicity experiment showed that PQ‐treated cells exhibited significant cell death, with the maximal toxicity occurring after 72 hours. However, the cells treated with Syzigium aromaticum were protected from the PQ‐induced cell death. Syzigium aromaticum exhibited rescue effect in proportion to its dose. The western blot analysis of PQ treated SH‐SY5Y cells demonstrated a dose‐dependent increase in the levels of p53, Setrin2, and phosphorylated AMPK (P‐AMPK). The levels of AMPK, mTOR and p70S6K remain unchanged, however, the phosphorylated p70S6K (P‐p70S6K) levels were declined. When cells were simultaneously treated with PQ and increasing dose of Syzigium aromaticum, the levels of p53, Sesn2, and P‐AMPK lowered similar to control. Further, there was an increase in the level of phosphorylated p70S6K (P‐p70S6K) in proportion to the dose of Syzigium aromaticum.ConclusionThe current study showed that the Syzigium aromaticum ethanolic extract prevented PQ‐induced cell death in SH‐SY5Y by targeting p53 and sestrin2.