摘要
目的:探讨对甲苯磺酰维达列汀(PV)对脂多糖(LPS)诱导的RAW264.7巨噬细胞焦亡和凋亡的影响,及其可能的抗炎机制。方法:MTT法检测RAW264.7的细胞活力;细胞流式术检测RAW264.7细胞的凋亡;Western blot检测caspase-3和caspase-1蛋白的表达;ELISA检测细胞培养上清液中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6浓度;RT-qPCR法检测NLRP3、caspase-1、GSDMD、IL-1β、IL-18、caspase-11、caspase-3和Bax mRNA的表达。结果:PV显著抑制LPS刺激的巨噬细胞的活力,且呈剂量依赖性,同时促进RAW264.7巨噬细胞的凋亡(P<0.01);显著减少巨噬细胞上清液释放TNF-α、IL-1β和IL-6(P<0.01);减少caspase-1并增加caspase-3蛋白的表达(P<0.01);下调细胞中NLRP3、caspase-1、GSDMD、IL-1β、IL-18和caspase-11并上调caspase-3和Bax mRNA的表达(P<0.01)。结论:PV可抑制LPS诱导的RAW264.7巨噬细胞发生细胞焦亡并促进其凋亡,使炎症因子释放减少。
AIM:To investigate effects of para-toluenesulfonyl vildagliptin(PV)on pyroptosis and apoptosis of RAW264.7 macrophages induced by lipopolysaccharide(LPS),and to explore its possible anti-inflammation mecha⁃nism.METHODS:The viability of RAW264.7 cells was measured by MTT assay.The apoptosis of RAW264.7 cells was analyzed by flow cytometry.The protein expression levels of caspase-3 and caspase-1 were determined by Western blot.The concentrations of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 were detected by ELISA.The mRNA expression levels of NLRP3,caspase-1,GSDMD,IL-1β,IL-18,caspase-11,caspase-3 and Bax were detect⁃ed by RT-qPCR.RESULTS:The viability of macrophages stimulated by LPS was inhibited significantly by PV in a dosedependent manner,and the apoptosis of macrophages was promoted at the same time(P<0.01).The contents of IL-1β,IL-6 and TNF-αin cell supernatant were decreased significantly(P<0.01).The protein expression of caspase-1 was de⁃creased but caspase-3 was increased(P<0.01).The mRNA expression of NLRP3,caspase-1,GSDMD,IL-1β,IL-18 and caspase-11 were down-regulated while caspase-3 and Bax mRNA were up-regulated(P<0.01).CONCLUSION:PV inhibits LPS-induced pyroptosis of RAW264.7 macrophages and promotes their apoptosis,resulting in reducing the re⁃lease of inflammatory cytokines.
作者
聂颖青
熊宝萍
刘海林
刘颖菊
NIE Ying-qing;XIONG Bao-ping;LIU Hai-lin;LIU Ying-ju(Department of Pharmacology,Chongqing Key Lab of Biochemistry and Molecular Pharmacology,Chongqing Medical University,Chongqing 400016,China;Department of Pharmacy,First People's Hospital of Chongqing Liangjiang New District,Chongqing 401121,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第4期702-710,共9页
Chinese Journal of Pathophysiology
基金
重庆市重庆市技术创新与应用发展专项面上项目(No.cstc2019jscx-msxmX0096)。