CD4^+T淋巴细胞在IgA肾病中致病作用机制研究  

The pathogenesis of CD4^+T lymphocytes in IgA nephropathy

作　　者：唐余燕 贺海东[1] 孙蔚倩[1] 张栋梁[1] 胡屏[1] 徐旭东[1] TANG Yu-yan;HE Hai-dong;SUN Wei-qian;ZHANG Dong-liang;HU Ping;XU Xu-dong(Department of Nephropathy,Minhang Branch Hospital affiliated to Fudan University(Department of Nephropathy,Minhang District Central Hospital),Shanghai 201199,China)

机构地区：[1]复旦大学附属闵行医院(上海市闵行区中心医院)肾脏科,上海201199

出　　处：《临床肾脏病杂志》2020年第2期147-153,共7页Journal Of Clinical Nephrology

基　　金：国家自然科学基金(81774060);上海市卫计委科研基金课题资助项目(20184Y0040);上海市闵行区自然科学研究课题资助项目(2018MHZ069)。

摘　　要：目的探讨CD4^+T淋巴细胞在IgA肾病(IgA nephropathy,IgAN)中致病作用机制。方法选取2017年8月至2018年8月于复旦大学附属闵行医院肾脏科住院治疗且经首次肾活检证实为IgAN患者作为IgAN组(45例),另外纳入我院体检中心体检健康者作为健康对照组(49例),采集健康对照组和IgAN组患者治疗前后的外周血液样本,分离制备外周血淋巴细胞、单核细胞及血清。采用ELISA检测外周血清CD4^+T淋巴细胞因子,包括γ干扰素(interferon-γ,IFN-γ)、白介素(interleukin,IL)-4、IL-17、IL-21、转化生长因子-β1(transforming growth factor-β1,TGF-β1)。采用流式细胞仪检测CD4^+T细胞的分布比例。采用RT-qPCR及Western blot检测JAK/STAT信号通路各信号分子的表达水平。结果IgAN组血清IL-4、IL-17、TGF-β1、IL-21水平高于健康对照组(P<0.05);而IFN-γ水平与健康对照组比较差异无统计学意义(P>0.05)。使用激素治疗1月后,IgAN组外周血清IL-4、IL-17、TGF-β1、IL-21水平均显著低于治疗前(P<0.01)。与健康对照组比较,治疗前IgAN组患者Th2及Treg细胞在T细胞中占比显著升高(P<0.05),而Th1的占比在两组受试者之中差异无统计学意义(P>0.05)。RT-qPCR及Western blot结果表明,治疗前IgAN组JAK1、JAK3、STAT3和STAT6表达水平显著高于正常对照组(P<0.05),而STAT5表达水平显著低于正常对照组(P<0.05),两组JAK2、TYK2、STAT1和STAT4表达水平比较差异无统计学意义(P>0.05)。结论CD4^+T细胞亚群失调可能参与IgAN患者的发病机制,与JAK/STAT信号通路表达失衡有关。因此,CD4^+T细胞因子(IL-4、IL-17、TGF-β1和IL-21)可能成为治疗IgAN的新靶点和监测IgAN病情的无创生物标志物。Objective To study the pathogenesis of CD4^+T lymphocytes in IgA nephropathy(IgAN).Methods A total of 45 cases of IgA nephropathy,hospitalized and diagnosed by first renal biopsy in Department of Nephropathy,Minhang Branch Hospital affiliated to Fudan University,were selected as the IgAN group(45cases).Another 49 healthy physical persons who examined in our medical center were selected as the normal control group(49 cases).The peripheral blood samples of the patients in the two groups before and after treatment were collected,to isolate peripheral blood lymphocytes,monocytes and serum.ELISA method was adopted to detect CD4^+T lymphocyte factor including interferon-γ(IFN-γ),interleukin 4(IL-4),IL-17,IL-21 and transforming growth factor-β1(TGF-β1).The distribution proportion of CD4^+T cells was detected by flow cytometry.Meanwhile,the expression level of JAK/STAT signal pathway was detected by RT-qPCR and Western blot.Results The levels of serum IL-4,IL-17,TGF-β1 and IL-21 in the IgAN group were higher than those in the normal control group(P<0.05);the IFN-γlevel had no statistical significant difference from that in the normal control group(P>0.05).After treatment for 1 month with hormone,the levels of serum IL-4,IL-17,TGF-β1and IL-21 in the IgAN group were all significantly lower than those before treatment(P<0.01).Compared with the normal control group,the proportions of Th2 and Treg cells before treatment in the IgAN group were increased significantly were increased significantly(P<0.05),but the proportion of Th1 had no statistically significant difference between the two groups(P>0.05).RT-qPCR and Western blot results showed,the expression levels of JAK1,JAK3,STAT3 and STAT6 before treatment in the IgAN group were significantly higher than those in the normal control group(P<0.05);while the STAT5 expression level was significantly lower than that in the normal control group(P<0.05).And,there was no significant difference in the expressions levels of JAK2,Tyk2,STAT1 and STAT4 between the two groups(P>0.0

关 键 词：IGA肾病 CD4^+T淋巴细胞 JAK/STAT信号通路 细胞因子 

分 类 号：R692.31[医药卫生—泌尿科学]