Abstract
Aim:
Accumulation of α-synuclein (α-syn) in the brain is a characteristic of Parkinson's disease (PD). In this study, we investigated whether treatment with tunicamycin, an endoplasmic reticulum (ER) stress inducer, led to the accumulation of α-syn in PC12 cells, and where α-syn protein was accumulated, and finally, whether bibenzyl compound 20c, a novel compound isolated from Gastrodia elata (Tian ma), could alleviate the accumulation of α-syn and ER stress activation in tunicamycin-treated PC12 cells.
Methods:
PC12 cells were treated with tunicamycin for different time (6 h, 12 h, 24 h, 48 h). Cell viability was determined by a MTT assay. Subcellular fractions of ER and mitochondria were extracted with the Tissue Endoplasmic reticulum Isolation Kit. The levels of α-syn protein and ER-stress-associated downstream chaperones were detected using Western blots and immunofluorescence.
Results:
Treatment of PC12 cells with tunicamycin (0.5–10 μg/mL) dose-dependently increased the accumulation of α-syn monomer (19 kDa) and oligomer (55 kDa), and decreased the cell viability. Accumulation of the two forms of α-syn was observed in both the ER and mitochondria with increasing treatment time. Co-treatment with 20c (10−5 mol/L) significantly increased the viability of tunicamycin-treated cells, reduced the level of α-syn protein and suppressed ER stress activation in the cells, evidenced by the reductions in phosphorylation of eIF2α and expression of spliced ATF6 and XBP1.
Conclusion:
Tunicamycin treatment caused accumulation of α-syn monomer and oligomer in PC12 cells. Bibenzyl compound 20c reduces the accumulation of α-syn and inhibits the activation of ER stress, which protected PC12 cells against the toxicity induced by tunicamycin.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (No 81274122, 81373997, 81273629, 81473376, U1402221, and 81573640), the National Mega-project for Innovative Drugs (No 2012ZX09301002-004), the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) (No IRT1007), Beijing Natural Science Foundation (No 7131013 and 7161011), and the Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study (No BZ0150).
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Mou, Z., Yuan, Yh., Lou, Yx. et al. Bibenzyl compound 20c protects against endoplasmic reticulum stress in tunicamycin-treated PC12 cells in vitro. Acta Pharmacol Sin 37, 1525–1533 (2016). https://doi.org/10.1038/aps.2016.75
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DOI: https://doi.org/10.1038/aps.2016.75
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