title: Rifampicin pre-treatment inhibits the toxicity of rotenone-induced PC12 cells by enhancing sumoylation modification of α-synuclein.
abstract: Our previous research revealed that rifampicin could protect PC12 (pheochromocytoma 12) cells from rotenone-induced cytotoxicity by reversing the aggregation of α-synuclein. Furthermore, increasing evidence indicated that the misfolded α-synuclein with SUMOylation, an important protein posttranslational modification, was easier to solubilize and was less toxic. Here, we investigated whether rifampicin could stabilize α-synuclein and prevent rotenone-induced PC12 cells from undergoing apoptosis by enhancing SUMOylation of α-synuclein. The expression of SUMO1 and SUMO2/3, the two main proteins responsible for the SUMOylation modification in PC12 cells, were detected by western blotting. Co-immunoprecipitation was performed to compare qualitatively the SUMOylation modification of α-synuclein. The cell viability and apoptosis rate were measured by a CCK-8 assay kit and flow cytometry, respectively. We targeted Ubc9 as a key enzyme in the SUMOylation modification pathway and knocked down the UBC9 gene using a short interfering RNA. Treatment with 150 μmol/L rifampicin, increased the expressions of SUMO1 and SUMO2/3 in cells by 1.5 times compared with the control group; meanwhile, the cell viability of rotenone-induced cells increased from 20 to 80% (P < 0.05). In addition, the increased SUMOylation activity in the cells stimulated by rifampicin was observed 18 h earlier compared with cells treated by rotenone alone. SUMOylation of α-synuclein was more significant in rifampicin-treated cells and Ubc9 upregulated cells. However, the same phenomenon and the protective effect of rifampicin were reversed after UBC9 knockout. In conclusion, rifampicin might reduce the cytotoxicity of rotenone-induced PC12 cells by promoting SUMOylation of α-synuclein.
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: D
authors.initials: D
authors.lastname: Lin
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: X
authors.initials: X
authors.lastname: Jing
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: M
authors.initials: M
authors.lastname: Lei
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: S
authors.initials: S
authors.lastname: Peng
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: T
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: D
authors.initials: D
authors.lastname: Zheng
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Zeng
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: X
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: L
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: S
authors.initials: S
authors.lastname: Xiao
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China.
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Neurology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, 107 Yanjiang West Road, Guangzhou 510080, China; Guangdong Province Key Laboratory of Brain Function and Disease, Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan 2nd Road, Guangzhou 510080, China. Electronic address: taoenxiang@163.com.
authors.forename: E
authors.initials: E
authors.lastname: Tao
mesh_terms: D000818:Animals; D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D000978:Antiparkinson Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016716:PC12 Cells; D010302:Parkinson Disease, Secondary / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D051381:Rats; D012293:Rifampin / Q000494:pharmacology*; D012402:Rotenone; D058207:Sumoylation / Q000187:drug effects*; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D000904:Antibiotics, Antitubercular; D000978:Antiparkinson Agents; D051844:alpha-Synuclein; D012402:Rotenone; D012293:Rifampin
keywords: PC12 cells; Parkinson's disease; Rifampicin; Rotenone; SUMOylation; α-synuclein