Abstract
Aim:
Our preliminary study shows that a bibenzyl compound isolated from Gastrodia elata, 2-[4-hydroxy-3-(4-hydroxybenzyl)benzyl]-4-(4-hydroxybenzyl)phenol (designated 20C), protects PC12 cells against H2O2-induced injury. In this study we investigated whether 20C exerted neuroprotective action in a cell model of Parkinson's disease.
Methods:
A cell model of Parkinson's disease was established in PC12 cells by exposure to rotenone (4 μmol/L) for 48 h. Cell viability and apoptosis were assessed, and intracellular ROS level and the mitochondrial membrane potential (MMP) were detected. The expression of apoptosis-related proteins Bax, Bcl-2, cytochrome c, cleaved caspase-3, and oxidative stress-related proteins Nrf2, HO-1 and NQO1 were examined using Western blotting. The mRNA levels of HO-1 and NQO1 were determined with RT-PCR. The nuclear translocation of Nrf2 was observed with immunofluorescence staining.
Results:
Treatment with rotenone significantly increased the number of apoptotic cells, accompanied by marked increases in the Bax/Bcl-2 ratio, cytochrome c release and caspase-3 activation. Rotenone also increased ROS accumulation, reduced MMP, and increased the nuclear translocation of Nrf2 as well as the mRNA and protein levels of the Nrf2 downstream target genes HO-1 and NQO1 in PC12 cells. Co-treatment with 20C (0.01–1 μmol/L) dose-dependently attenuated rotenone-induced apoptosis and oxidative stress in PC12 cells. Nrf2 knockdown by siRNA partially reversed the protective effects of 20C in rotenone-treated PC12 cells.
Conclusion:
The bibenzyl compound 20C protects PC12 cells from rotenone-induced apoptosis, at least in part, via activation of the Nrf2/ARE/HO-1 signaling pathway.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (No 81274122, No 81102831, and No 81273629), the National Key Sci-Tech Major Special Item (No 2012ZX09301002-004 and No 2012ZX09103101-006), the National High-Tech R&D Programme (863 Program) (No 2012AA020303), the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) (No IRT1007), the Specialized Research Fund for the Doctoral Program of Higher Education of China (No 20121106130001), Beijing Natural Science Foundation (No 7131013 and No 7142115), Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study (No BZ0150).
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Huang, Jy., Yuan, Yh., Yan, Jq. et al. 20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway. Acta Pharmacol Sin 37, 731–740 (2016). https://doi.org/10.1038/aps.2015.154
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DOI: https://doi.org/10.1038/aps.2015.154
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