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  

作　　者：Ju-yang HUANG Yu-he YUAN Jia-qing YAN Ya-nan WANG Shi-feng CHU Cheng-gen ZHU Qing-lan GUO Jian-gong SHI Nai-hong CHEN 

机构地区：[1]State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China [2]College of Pharmacy, Hunan University of Chinese Medicine, Changsha 410000, China

出　　处：《Acta Pharmacologica Sinica》2016年第6期731-740,共10页中国药理学报（英文版）

摘　　要：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 H202-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 pmol/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 NQ01 were examined using Western blotting. The mRNA levels of HO-1 and NQ01 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 NQ01 in PC12 cells. Co-treatment with 20C （0.01-1 pmol/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.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 H202-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 pmol/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 NQ01 were examined using Western blotting. The mRNA levels of HO-1 and NQ01 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 NQ01 in PC12 cells. Co-treatment with 20C （0.01-1 pmol/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.

关 键 词：bibenzyl compound Gastrodia elata ROTENONE apoptosis oxidative stress Nrf2 NEUROPROTECTION Parkinson＇s disease PC12 cells 

分 类 号：Q517[生物学—生物化学] Q421