【摘要】 目的研究细胞因子信号转导抑制因子3(SOCS3)在风湿性心脏病大鼠脾脏CD4~+T细胞分化中的作用及机制。方法建立灭活A组链球菌(GSA)诱导的大鼠风湿性心脏病模型,分离脾脏CD4~+T细胞;实验分6组:空白对照组(A组)、GSA免疫大鼠CD4~+T细胞组(B组)、GSA免疫大鼠CD4~+T细胞+PYrAd-rSOCS3转染组(C组)、GSA免疫大鼠CD4~+T细胞+PYrAd-GFP转染组(D组)、GSA免疫大鼠CD4~+T细胞+siRNA-SOCS3转染组(E组)、GSA免疫大鼠CD4~+T细胞+siRNA-control转染组(F组);采用RT-qPCR和Western blot法检测SOCS3、P/T-STAT3(仅检测蛋白)、T-bet、Gata3、RORγt和Foxp3的mRNA和蛋白表达;采用ELISA法检测各组细胞上清液中细胞因子IFN-γ、IL-4、IL-17和TGF-β的水平。结果与A组比较,B组SOCS3、STAT3、P-STAT3、T-bet、RORγt、IFN-γ和IL-17表达上调(均P<0.05),而Gata3、Foxp3、IL-4和TGF-β表达下调(均P<0.05)。与B组比较,C组SOCS3表达进一步上调(P<0.01),而STAT3、P-STAT3、T-bet、RORγt、IFN-γ和IL-17表达下调(均P<0.05),同时Gata3、Foxp3、IL-4和TGF-β表达上调(均P<0.05)。与B组比较,E组SOCS3表达下调(P<0.01),STAT3、P-STAT3、T-bet、RORγt、IFN-γ和IL-17表达上调(均P<0.05),而Gata3、Foxp3、IL-4和TGF-β表达下调(均P<0.05)。结论 SOCS3通过抑制STAT3的磷酸化而抑制CD4~+T细胞向Th1和Th17细胞过度极化,在风湿性心脏病发病中可能具有负向调节的作用。更多还原

【Abstract】 Objective To study the role and mechanism of suppressor of cytokine signaling 3(SOCS3)in the differentiation of CD4~+T lymphocytes in rats with rheumatic heart disease.Methods A rat model of rheumatic heart disease was established by using group A streptococcal(GSA),and spleen CD4~+T lymphocytes were isolated.The experiment was divided into six groups:control(group A),immunization with GSA(group B),immunization with GSA and transfection with SOCS3 recombinant adenovirus(PYrAd-rSOCS3)(group C),immunization with GSA and transfection with PYrAd-GFP(group D),immunization with GSA and transfection with siRNA-SOCS3(group E),and immunization with GSA and transfection with siRNA-control(group F).The expression of SOCS3,P/-STAT3,T-bet,Gata3,RORγt and Foxp3 was detected by RTqPCR and Western Blot.The levels of IFN-γ,IL-4,IL-17 and TGF-βin the supernatant were measured by ELISA.Results Compared with group A,the expression of SOCS3,STAT3,PSTAT3,T-bet,RORγt,IFN-γand IL-17 was up-regulated while the expression of Gata3,Foxp3,IL-4 and TGF-βwas down-regulated in group B(P<0.05).Compared with group B,the expression of SOCS3,Gata3,Foxp3,IL-4 and TGF-βwas increased but the expression of STAT3,PSTAT3,T-bet,RORγt,IFN-γand IL-17 was decreased in group C(P<0.05 or P<0.01).In addition,compared with group B,the expression of SOCS3,Gata3,Foxp3,IL-4 and TGF-βwas decreased but the expression of STAT3,P-STAT3,T-bet,RORγt,IFN-γand IL-17 was increased in group E(P<0.05 or P<0.01).Conclusion SOCS3 may play a negative regulatory role in the pathogenesis of rheumatic heart disease through the inhibition of STAT3 phosphorylation and excessive polarization of CD4~+T lymphocytes toward Th1 and Th17 cells.更多还原