TNFα通过TNFR1/PKCα和TNFR2信号通路诱导人肾小球系膜细胞IP_3R1的表达  

TNFα induces IP_3R1 expression via TNFR1/PKCα and TNFR2 signaling pathways in human mesangial cells

作　　者：王育蓉[1] 章欢[1] 孙辉[1] 李翠红[1] 刘沛[2] 

机构地区：[1]江西省南昌大学附属医院九江市第一人民医院消化科,江西省九江市332000 [2]中国医科大学附属一院肝病中心,辽宁省沈阳市110001

出　　处：《世界华人消化杂志》2013年第6期521-526,共6页World Chinese Journal of Digestology

基　　金：江西省卫生厅科研基金资助项目;No.20123192~~

摘　　要：目的:探讨肿瘤坏死因子α(tumor necrosis factor α,TNFα)对人肾小球系膜细胞(human mesangial cells,HMCs)IP3R1蛋白和mRNA表达的影响及PKC和肿瘤坏死因子受体(tumornecrosis factor receptor,TNFR)在此信号通路中的作用,以期为肝肾综合征(hepatorenal syndrome,HRS)肾小球滤过率下降的发生机制和防治思路提供理论依据.方法:用实时定量PCR和免疫印记法检测IP3R1mRNA和蛋白表达的情况,并分别用PMA耗竭内源性蛋白激酶C(endogenous protein kinase C,PKC),PKCα特异性抑制剂Safingol和PKCδ特异性抑制剂Rottlerin及HA-DN-PKCα质粒转染干预上述诱导实验.同时,用免疫印记法检测TNFα对PKCα和p-PKCα的表达的影响.此外,免疫印记法检测TNFR对PKCα的活化及IP3R1蛋白表达的影响.结果:TNFα作用HMCs后,IP3R1蛋白和mRNA表达均明显增加.PMA、PKCα抑制剂Safingol,HA-DN-PKCα质粒转染均能明显阻断TNFα诱导的IP3R1蛋白的上调,而P K Cδ抑制剂Rottlerin无明显阻断作用.TNFα刺激HMCs,各不同时间组总PKCα蛋白表达无明显差异,而TNFα刺激8h p-PKCα蛋白表达明显增加.TNFR1抗体+TNFα组和TNFR2抗体+TNFα组,IP3R1蛋白表达均有明显的不同程度的下降.TNFR1抗体+TNFα组p-PKCα蛋白表达明显下降,而TNFR2抗体+TNFα组,p-PKCα蛋白表达无明显变化.结论:TNFα能上调HMCs中IP3R1蛋白及IP3R1mRNA的表达.活化的PKCα可能在TNFα上调IP3R1的表达中起重要作用.TNFα可能通过TNFR1/PKCα途径及TNFR2途径上调IP3R1的表达.AIM： To investigate the role of PKC and TNFR in TNFα-induced expression of IP3R1 in human mesangial cells （HMCs）. METHODS： Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effect of TNFα treatment on IP3R1 mRNA and protein expression. Depletion of PKC with PMA, treatment with PKC kinase inhibitors, and overexpression of dominant negative mutant of PKCα were used to examine the role of PKC in TNFα-induced expression of IP3R1 in HMCs. The expression of total PKCα and p-PKCα was assayed by Western blot. The contribution of TNFR1 and TNFR2 to PKCα activation and TNFα-induced IP3R1 expression was also detected by Western blot. IRESUKTS ： Treatment with TNFα increased IP3R1 mRNA and protein expression in HMCs, and this effect could be blocked by prolonged incubation with PMA, Safingol treatment and transfection with domain negative PKCα construct. TNFα could promote PKCα autophosphorylation. Both anti-TNFR1 and TNFR2 antibodies attenuated TNFα-induced IP3R1 expression, while only anti-TNFR1 antibody attenuated TNFα-induced PKCα activity. CONCLUSION： TNFα increases the expression of IP3R1 through the TNFR1/PKCα and TNFR2 signaling pathways in HMCs.

关 键 词：肝肾综合征 肿瘤坏死因子α Ⅰ型1 4 5-三磷酸肌醇受体 蛋白激酶CΑ 肿瘤坏死因子受体 

分 类 号：R575.2[医药卫生—消化系统]