换一批细胞因子信号转导抑制因子3过表达对IgA肾病患者IgA1刺激人肾小球系膜细胞增殖的抑制作用
Over-expression of suppressor of cytokine signaling 3 inhibits the proliferation of human mesangial cells stimulated by aggregated IgA1 from IgA nephropathy patients
摘要目的 探讨细胞因子信号转导抑制因子3(SOCS3)对IgA肾病(IgAN)患者血清IgA1刺激诱导人肾小球系膜细胞(HMC)增殖的作用及其可能机制.方法 采用Jacalin亲和层析联合丙烯葡聚糖凝胶S-200凝胶过滤法纯化聚合IgAN患者血清热聚合IgA1 (aIgA1).以腺病毒为载体转染SOCS3至体外培养的HMC,再以aIgA1刺激培养12 ～ 48 h,分为空白对照组、IgA1组、IgA1+空载腺病毒组、IgA1+SOCS3腺病毒组.采用噻唑蓝(MTT)法检测系膜细胞增殖情况,Western印迹法和实时定量PCR法检测各组SOCS3、Toll样受体4(TLR4)、转化生长因子β1(TGF-β1)蛋白和mRNA表达水平.结果 aIgA1刺激HMC 24 h后开始表现出促增殖作用,与空白对照组相比,aIgA1刺激后SOCS3、TLR4、TGF-β1蛋白和mRNA表达均有显著升高(P＜0.05).与IgA1组、IgA1+空载腺病毒组相比,IgA1 +SOCS3腺病毒组TLR4、TGF-β1蛋白和mRNA表达均显著下降(P＜0.05),HMC增殖程度显著降低(P＜0.05).结论 上调SOCS3表达可降低aIgA1刺激后HMC的TLR4和TGF-β1表达水平,并抑制aIgA1诱导的HMC细胞增殖.
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abstractsObjective To investigate the effect of suppressor of cytokine signaling 3 (SOCS3)on the proliferation of human mesangial cells stimulated by aggregated IgA1 (aIgA1) from patients with IgA nephropathy(IgAN),and explore its possible mechanism.Methods Serum monomeric IgA1 was isolated with jacalin affinity and Sephacryl S-200 HR chromatography from IgAN patients,and then heated to aggregated form (aIgA1).Human glomerular mesangial cells(HMC) were transfected with AdvSOCS3-IRES2-EGFP for 48 hours,and incubated with aIgA1 for 12-48 h.The cells were divided into blank control group,IgA1 group,IgA1 +Adv-EGFP group and IgA 1 +Adv-SOCS3-IRES2-EGFP group.The mesangial cell proliferation was observed through MTT,and the levels of SOCS3,TLR4,TGF-β1 protein and mRNA were detected through Western blotting and real-time PCR.Results HMC proliferation was promoted significantly after IgA1 stimulated at 24 h.Compared with control group,the protein and mRNA expression of SOCS3,TLR4,TGF-β1 were significantly increased in IgA1 group (P ＜ 0.05).Compared with IgA1 group and IgA1 +Adv-EGFP group,MTT absorbency was obviously reduced after incubation with aIgA1 for 24 h and 48 h in IgA+Adv-SOCS3-IRES2-EGFP group,and the protein and mRNA expression of TLR4 and TGF-β1 were significantly decreased in IgA1 +AdvSOCS3-EGFP group (P＜ 0.05).Conclusion Over-expression of SOCS3 may inhibit the proliferation of HMC stimulated by aIgA1,partly through down-regulating the expression of TLR4 and TGF-β1.
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