The mechanism of HDAC6 in paraquat-induced autophagy dysfunction of dopaminergic neurons by mediating aggresome-autophagy-lysosomal pathway
10.3760/cma.j.cn121094-20201221-00705
- VernacularTitle:组蛋白去乙酰化酶6介导蛋白包涵体-自噬-溶酶体途径在百草枯诱导多巴胺能神经元自噬障碍中的作用
- Author:
Weiguang YAN
1
;
Yifan WANG
;
Tian TIAN
;
Kaidong WANG
;
Baofu ZHANG
;
Bingyang ZHANG
;
Min HUANG
Author Information
1. 750004 银川，宁夏医科大学公共卫生与管理学院职业卫生与环境卫生学系，宁夏环境因素与慢性病控制重点实验室
- Keywords:
Paraquat;
Autophagy;
HDAC6;
Aggresome-autophagy-lysosome pathway
- From:
Chinese Journal of Industrial Hygiene and Occupational Diseases
2021;39(4):241-247
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of HDAC6 mediated aggresome-autophagy-lysosome pathway in paraquat-induced autophagy in dopaminergic neurons.Methods:Human neuroblastoma cell (SH-SY5Y cell) was used as model of dopaminergic neurons in vitro. The cells were treated with terminal concentrations of 0, 25, 50, 100, 200 and 400μmol/L PQ for 24 hours, and the cells were induced by 100 μmol/L PQ for different time (0, 12, 24, 36, 48, 60 and 72 h) . Cell viability was detected by CCK-8 assay. The expression levels of HDAC6, α-syn, Dynein IC1/2, LC3, Beclin1, p62 and Lamp-1 were detected by Western blot. Immunofluorescence double-labeling method was used to observe the expression and localization of HDAC6, α-syn, Dynein IC1/2, LC3, Lamp-1 and γ-tubulin in cells.Results:CCK-8 assay showed PQ induced cell survival rate decrease in a time and dose dependent manner ( R=-0.950、-0.960, P<0.05) .Western blot showed that compared with control group, the protein levels of HDAC6, α-syn, p62 in PQ-exposed group were significantly increased ( P<0.05) , but there was a significant decrease in expression level of the ratio of autophagy-related protein LC3 Ⅱ/LC3 Ⅰ, Beclin1, Dynein IC1/2, Lamp-1in PQ-exposed group ( P<0.05) . The results of immunofluorescence double-labeling showed that compared with the control group, the fluorescence signals of HDAC6 and α-syn in the PQ-exposed group increased, and the protein expression level increased, while the fluorescence signals of Dynein IC1/2, LC3, and Lamp-1 decreased. The protein expression level is reduced. HDAC6 gradually accumulates from the diffuse shape to the nucleus; Under normal circumstances, α-syn, Dynein IC1/2, γ-tubulin, LC3, and Lamp-1 are mainly distributed in the cytoplasm. After PQ is infected, they gather in the nucleus and co-localize with HDAC6 in the area around the nucleus. Conclusion:PQ may induce abnormal aggregation of α-syn by inducing HDAC6-mediated aggresome-autophagy-lysosomal pathway disorder.
