HMGB1、TNFAIP3蛋白对狼疮性肾炎系膜细胞增殖的影响及其NF-κB信号通路机制

Analysis of effect of HMGB1 and TNFAIP3 on proliferation of mesangial cells in lupus nephritis and the mechanism NF-κB signaling pathway

目的基于核因子-κB(NF-κB)信号通路研究高迁移率族蛋白1(HMGB1)、肿瘤坏死因子α诱导蛋白3(TNFAIP3)对狼疮性肾炎(LN)肾小球系膜细胞(MC)增殖的影响。方法分离、培养10只MRL-Faslpr/JNju小鼠(LN小鼠)的MC,将其分为空白对照组、溶剂对照组、HMGB1干预组。空白对照组不作任何处理,溶剂对照组加入200μg/L甲醇溶剂,HMGB1干预组加入200μg/L人重组HMGB1蛋白。采用CCK8法检测12、24、36、48 h时3组肾小球系膜细胞增殖率。采用免疫印迹法(Western Blotting)检测3组MC中HMGB1、TNFAIP3、NF-κB、NF-κB抑制蛋白α(IκB-α)蛋白表达量。结果12、24、36、48 h时,HMGB1干预组MC增殖率显著高于空白对照组、溶剂对照组(P<0.05);HMGB1干预组MC增殖率随时间延长而显著升高(P<0.05);HMGB1干预组MC中HMGB1、TNFAIP3、NF-κB蛋白表达量显著高于空白对照组、溶剂对照组(P<0.05);干预组MC中IκB-α蛋白表达量显著低于空白对照组、溶剂对照组(P<0.05);溶剂对照组MC增殖率及HMGB1、TNFAIP3、NF-κB、IκB-α蛋白表达量较空白对照组差异无统计学意义(P>0.05)。结论HMGB1可能通过上调TNFAIP3表达,诱发NF-κB信号通路活化,介导LNMC过度增殖,这可为LN的发病机制研究和靶向治疗提供理论基础。

Objective To investigate the effect of high mobility group box-1 protein(HMGB1)and tumor necrosis factor alpha induced protein 3(TNFAIP3)on proliferation of mesangial cells in lupus nephritis(LN)based on nuclear transcription factor-κB(NF-κB)signaling pathway.Methods After isolation and culture,the glomerular mesangial cells of 10 MRL-Faslpr/JNju mice were divided into blank control group,solvent control group and HMGB1 intervention group.The blank control group received no treatment,the solvent control group was added with 200 g/L methanol solvent,and the HMGB1 intervention group was added with 200 g/L human recombinant HMGB1 protein.CCK8 method was used to detect the proliferation rate of glomerular mesangial cells in each group at 12 h,24 h,36 h,48 h,and Western Blot was used to detect the expression levels of HMGB1,TNFAIP3,NF-κB and inhibitor of NF-kB,αisoform(IκB-α)in each group.Results The proliferation rate of mesangial cells in HMGB1 intervention group was significantly higher than that in blank control group and solvent control group at 12 h,24 h,36 h and 48 h(P<0.05).The proliferation rate of mesangial cells in HMGB1 group increased significantly with intervention time(P<0.05).The expression levels of HMGB1,TNFAIP3,and NF-κB proteins in the glomerular mesangial cells of the HMGB1 intervention group were significantly higher than those of the blank control group and the solvent control group(P<0.05);The expression of IκB-αprotein in mesangial cells of intervention group was significantly lower than that of blank control group and solvent control group(P<0.05).There was no significant difference in the proliferation rate of mesangial cells and the expression of HMGB1,TNFAIP3,NF-κB,IκB-αprotein between blank control group and solvent control group(P>0.05).Conclusion The mediation of HMGB1 on the excessive proliferation of mesangial cells in lupus nephritis may be achieved through the increase of TNFAIP3 expression and the activation of NF-κB signaling pathway,which can provide a theoretical basis for the pathogenesis research and targeted treatment of lupus nephritis.