Role of NLRP3-dependent activation of caspase-1 in mediating neuroinflammatory response in a mouse model of MPTP-induced Parkinson's disease
10.3760/cma.j.issn.0254-5101.2018.06.005
- VernacularTitle:Caspase-1通过炎症小体激活MPTP诱导的PD小鼠神经炎症反应的相关研究
- Author:
Ling YU
1
;
Hui LIANG
;
Min KONG
Author Information
1. 264000,烟台市烟台山医院神经内科
- Keywords:
NLRP3;
Caspase-1;
SH-SY5Y cell;
Parkinson's disease
- From:
Chinese Journal of Microbiology and Immunology
2018;38(6):427-433
- CountryChina
- Language:Chinese
-
Abstract:
Objective To understand the role of NOD-like receptor pyrin domain containing 3 (NLRP3)-dependent activation of cysteinyl aspartate specific proteinase 1 (caspase-1) in mediating the neuroinflammatory response in mice with MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)-induced Parkinson's disease(PD) and to investigate the underlying mechanism. Methods Male C57BL/6 mice were randomly divided into two groups:experimental and control groups. MPTP solution (30 mg·kg-1·d-1) was given to mice through intraperitoneal injection to prepare the PD model and equal amount of saline was used to set up the control group. Changes in mouse behavior were observed through pole climbing and suspension experiment. Expression of inflammasome in the midbrain of mice was detected by Western blot. SH-SY5Y cells,a human neuroblastoma cell line,were cultured in vitro and randomly divided into five groups including control,nonsense,siRNA-NLRP3,MPP+(1-methyl-4-phenylpyridinium) and siRNA+MPP+groups. siRNA-NLRP3 was transfected into SH-SY5Y cells using Lipofectamine 2000 to silence the expression of NLRP3 gene. Then the transfected cells were incubated with MPP+for 48 h to observe the protective effect of NLRP3 on nerve cells. MTT assay and flow cytometry were performed to measure the viability and the apoptosis rate of SH-SY5Y cells,respectively. Western blot was used to detect the levels of NLRP3,B-cell lymphoma 2 (Bcl-2),Bcl-2-associated X protein (Bax) and cleaved caspase-3. Results Compared with the control group,the mice in the experimental group spent more time climbing from the top to the bottom of the pole (P<0. 05),got a lower score on suspension experiment (P<0. 05) and showed enhanced expression of NLRP3, IL-1β,Pro-IL-1β and caspase-1 in the substantia nigra (P<0. 05). Compared with the MPP+group,the siR-NA+MPP+group showed significantly inhibited expression of NLRP3, cleaved caspase-3 and Bax ( P<0. 05),enhanced cell viability (P<0. 05),suppressed cell apoptosis (P<0. 05) and promoted expression of Bcl-2 ( P<0. 05). Conclusion NLRP3-dependent activation of caspase-1 plays an important role in the nerve injury in PD. Inhibition of NLRP3 expression in vitro can significantly enhance the vitality and inhibit the apoptosis of SH-SY5Y cells,which may have a protective effect on nerve cells through regulating mito-chondrial apoptosis signaling pathway and provide a new drug target for the treatment of PD.
