Objective: The study analyzed the neuroprotective effect of ethanolic extract of clove against Parkinson’s disease (PD) type toxicity in SHSY5Y cell line model and interrogation of underlying molecular mechanism.

Background: Although the exact causes of sporadic PD remain unclear, numerous environmental risk factors, especially neurotoxins, have been implicated in its aetiology.  The national Institute on Aging in the National Institutes of Health (NIH) has reported a common spice, clove with the highest ORAC (Oxygen Radical Absorbance Capacity) value. Clove stimulates nerves by pacifying Vata, one of the three aspects of healthy aging.

Method: The ethanolic extract of clove was prepared using the cold extraction process by crushing, soaking in ethanol, and stirred for 72 h followed by filtration and evaporation to remove complete ethanol. The PD type toxicity was induced with different concentrations of paraquat (PQ) in SH-SY5Y cells for 24, 48 and 72 h and cytotoxicity assessment was done by MTT assay. The rescue effect of clove from PQ induced toxicity was performed by simultaneous treatment of cells with 100μM PQ and different concentrations of clove followed by MTT assay. To elucidate the underlying mechanism, the protein expression analysis was performed using immunoblotting.

Results: The toxicity assay of PQ revealed considerable cell death in PQ-treated cells with maximum toxicity found at 72h. However, the clove treated cells were able to protect themselves from neuronal cell death upon exposure to PQ.  The rescue effect was mediated in a dose-dependent manner of clove. The immunoblot analysis revealed the elevated level of p53 and Setrin2 and phosphorylated AMPK (P-AMPK) (Thr172) in PQ treated SH-SY5Y cells in a dose-dependent manner. The levels of AMPK, mTOR, and p70S6K were unaltered. Interestingly, the phosphorylated p70S6K (P-p70S6K) (Thr389) decreased in a dose-dependent manner of PQ. However, upon simultaneous treatment of cells with 100 µM of PQ and increasing dose of clove, p53, Sesn2, and P-AMPK declined to the level of control. Also, the level of phosphorylated p70S6K (P-p70S6K) (Thr389) increased with an increasing dose of clove.

Conclusion: The present study reported that ethanolic extract of clove abrogated the PQ-induced cell death of SH-SY5Y by lowering the oxidative and ER stress and boosting the antioxidant system involving P53 and Sestrin2.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/clove-mediated-rescue-in-parkinsons-disease-involves-modulation-of-p53-and-sestrin2-in-shsy5y-cell-line-model/