外源性硫化氢对肝细胞NLRP3炎症小体的影响  

Effect of exogenous hydrogen sulfide on expression of NLRP3 inflammasome in hepatocytes

作　　者：王红钢 孙伟力 钟培育 吴东栋 王军 蔡欢[2] 王国英 乔玲 张涛 李彦章 WANG Honggang;SUN Weili;ZHONG Peiyu;WU Dongdong;WANG Jun;CAI Huan;WANG Guoying;QIAO Ling;ZHANG Tao;LI Yanzhang(The Basic Medical College of Henan University,Kaifeng 475004, China;Henan University Minsheng College, Kaifeng 475004, China)

机构地区：[1]河南大学基础医学院,河南开封475004 [2]河南大学民生学院,河南开封475004

出　　处：《中国病理生理杂志》2018年第1期118-122,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31300841);河南省高校重点科研项目(No.16A310001);河南大学2016年度大学生创新性实验计划(No.16NB047)

摘　　要：目的:研究外源性硫化氢(H_2S)对人肝细胞NLRP3炎症小体的影响。方法:采用不同浓度的脂多糖(LPS)诱导人肝细胞L02和SMMC-7721建立炎症模型,Western blot检测细胞中NLRP3炎症小体的表达并结合细胞毒性实验(MTT法)确定合适的LPS浓度。细胞分为4组:对照组用普通培养基培养18.5 h;LPS组用普通培养基培养0.5 h后,再用100μg/L LPS刺激18 h;LPS+H_2S组和H_2S组用200μmol/L硫氢化钠(Na HS)刺激0.5 h后,再分别用100μg/L LPS和普通培养基培养18 h。各组处理后分别收集细胞,Western blot检测细胞中NLRP3和caspase-1的蛋白表达量。结果:与对照组相比,LPS组细胞内NLRP3和caspase-1的表达增加(P<0.05),H_2S组细胞内的NLRP3和caspase-1表达无明显变化;与LPS组相比,LPS+H_2S组细胞内的NLRP3和caspase-1表达减少(P<0.05)。结论:外源性H_2S可抑制人肝细胞中NLRP3炎症小体的表达。AIM:To evaluate the effect of exogenous hydrogen sulfide(H2S)on the expression of NLRP3inflammasome in hepatocytes.METHODS:The hepatocytes L02and SMMC-7721were used to establish the model of inflammation by stimulating with lipopolysaccharide(LPS)at different concentrations in vitro.The expression of NLRP3inflammasome in the hepatocytes was detected by Western blot and the cell viability was measured by MTT assay for determining appropriate concentration of LPS.The hepatocytes were divided into4groups:the cells in control group were incubated with normal medium for18.5h;the cells in LPS group were incubated with normal medium for0.5h followed by100μg/L LPS for18h;the cells in LPS+H2S group and H2S group were incubated with200μmol/L sodium hydrosulfide hydrate(NaHS)for0.5h followed by100μg/L LPS or normal medium for18h,respectively.The protein expression of NLRP3and caspase-1in the cells of every group was determined by Western blot.RESULTS:Compared with control group,the protein expression of NLRP3and caspase-1increased significantly in LPS group(P<0.05)and had no significant change in H2S group.Compared with LPS group,the protein expression of NLRP3and caspase-1in LPS+H2S group decreased significantly(P<0.05).CONCLUSION:In hepatocytes,exogenous H2S suppresses the expression of NLRP3inflammasome.

关 键 词：硫化氢 NLRP3炎症小体 肝细胞 脂多糖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R363[医药卫生—基础医学]