Rifampin protects Parkinson's disease model cell by promoting autophagy
WANG Chun-yan 1, FENG Fan 1,2, GONG Qi-xia 2, SHU Yu 2, WAN Sheng-xia 2,ZHAO Jing 2, XU Ping 2, QIAN Jin-jun 2
1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. Department of Neurology, the Fourth Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001, China)
Abstract：Objective: To investigate the effect of rifampicin on the level of α-synuclein(α-syn) multimer and its relationship with autophagy. Methods: Cells were treated with different concentrations of rotenone, rifampicin, autophagy inhibitor chloroquine and autophagy promoter rapamycin, MTT assay was used to detect the cell viability, and to screen the best experimental concentration. After cells were treated with chloroquine and rapamycin at different time points, Western blotting was used to detect the expression of Beclin-1 and α-syn multimer. SH-SY5Y cells were divided into four groups: control group, rotenone group, rifampicin+rotenone group and rifampicin group. The apoptosis rate of each group was detected by flow cytometry. Expression of the α-syn multimer and autophagy marker proteins Beclin-1, LC3-Ⅱ/I and p62 were detected by Western blotting. On the basis of the abovementioned grouping, Western blotting was used to detect the α-syn multimer expression of each group after pretreatment with chloroquine or rapamycin. Results: The cell survival rate was the highest when the concentration of rifampicin was 150 μmol/L. The optimal treatment conditions for chloroquine and rapamycin to regulate autophagy were 10 μmol/L(1 h) and 10 nmol/L(2 h), respectively. Compared with the control group, rotenone and chloroquine significantly increased α-syn multimer expression(P<0.01) and decreased Beclin1 expression level(P<0.05). Rapamycin increased Beclin-1 expression level(P<0.01). Compared with the control group, the rotenone group showed higher apoptosis rate(P<0.05) and higher expression of p62 and α-syn multimer, while with lower levels of Beclin-1 and LC3-Ⅱ/Ⅰ(P<0.01). Compared with the rotenone group, the rifampicin group showed lower apoptosis rate (P<0.05) and lower expression of p62 and α-syn multimer, Beclin-1 level and LC3-Ⅱ/Ⅰ(P<0.01). The expression of α-syn multimer was increased after chloroquine treatment compared with rifampicin treatment group(P<0.01). Conclusion: Rifampicin reduces rotenoneinduced apoptosis in dopaminergic neurons and may reduce α-syn multimer levels by promoting autophagy.
［Key words］Rifampin; Parkinsons disease; αsynuclein; autophagy
2018, Vol. 28 
