α-synuclein activates NLRP3 inflammatory bodies to mediated nerve cell pyroptosis

doi:10.12092/j.issn.1009-2501.2019.06.006

Abstract

Abstract:

AIM: To study the effect and mechanism of α-synuclein on nerve cell pyroptosis. METHODS: HT22 cell was cultured in vitro and interfered by α-synuclein. Cell viability was measured by CCK-8 method and IC50 value was determined. After setting up the control group (Con group), TUNEL staining was used to detect the pyroptosis level of cells, immunofluorescence was used to observe the expression of GSDMD-N, Western blot was used to detect the expression levels of NLRP3, ASC, GSDMD, GSDMD-N and Caspase-1 in cells, and enzyme-linked immunosorbent assay was used to detect the expression levels of IL-18 and IL-1β in culture medium. After HT22 was pretreated with Caspase-1 inhibitor, the cells were also treated with α-synuclein of IC50 value, and the α-synuclein group (only α-synuclein intervention) and α-synuclein+ML132 group were set up to detect the level of pyroptosis. RESULTS:The IC50 value of α-synuclein was 50 nmol/L. After the intervention of α-synuclein, the number of nerve cells increased significantly. TUNEL staining showed that the number of positive cells was more than that of Con group. Immunofluorescence staining showed that the expression level of GSDMD-N in cells was up-regulated. The expression levels of NLRP3, ASC, GSDMD-N and Caspase-1 in cells were significantly higher than those in Con group, while the level of GSDMD was down-regulated in cell culture medium. The levels of IL-18 and IL-1β were up-regulated. After treatment with Caspase-1 inhibitor, the pyroptotic level of 50 nmol/L α-synuclein was significantly lower than that of α-synuclein group. Immunofluorescence staining showed that the level of GSDMD-N was down-regulated. The expression levels of NLRP3, ASC, GSDMD-N and Caspase-1 in cells were significantly lower than those in α-synuclein group, while the level of GSDMD was up-regulated. The levels of IL-18 and IL-1β in culture medium were down regulation. CONCLUSION:α-synuclein may play a role in the development of Parkinson's disease by activating NLRP3 inflammatory bodies.

Key words: pyroptosis, α-synuclein, inflammatory bodies, GSDMD

CLC Number:

HAN Chenyang, ZHANG Xiaoling, YANG Yi, GUO Li, GUAN Qiaobing. α-synuclein activates NLRP3 inflammatory bodies to mediated nerve cell pyroptosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(6): 637-643.

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