大鼠系膜增生性肾小球肾炎外周血Th17、Treg及炎症因子演变特征

The study on the evolution characteristics of peripheral blood Th17,Treg and inflammatory factors in rats with mesangial proliferative glomerulonephritis

目的研究抗Thy1.1致系膜增生性肾小球肾炎(MsPGN)模型大鼠外周血辅助性T细胞17(Th17)、调节性T细胞(Treg)与炎症因子演变特点。方法取清洁级雄性SD大鼠36只,随机分为正常组与MsPGN组(n=18);采用大鼠单次尾静脉注射1.29 mg/kg Anti-Rat CD90(Thy1.1)诱导MsPGN模型,于3、7和14 d随机分别选取6只大鼠进行肾组织HE、PAS染色和IL-17A、Foxp3免疫组化分析;测定24 h尿蛋白、血尿素与肌酐含量;流式细胞术检测外周血Th17、Treg的组成变化;ELISA检测血清IL-17A、IL-6、IL-10和TGF-β1含量。结果与正常组相比,MsPGN大鼠肾组织HE和PAS染色在肾皮质部分的肾小球系膜区均出现系膜细胞及系膜基质增生,7 d达峰;免疫组化结果显示在肾皮质部分的肾小球系膜区IL-17A明显升高,3 d达峰(P<0.05),Foxp3在3 d较正常组降低后升高,7 d达峰(P<0.05);24 h尿蛋白和血尿素在3个时间点含量均升高,3 d达峰(P<0.05),而血肌酐变化差异无统计学意义;流式细胞术检测结果表明不同时间点的Th17均升高,3 d达峰,而Treg在不同时间点均低于对照,但整体变化呈升高趋势(P<0.05)。ELISA结果表明IL-6、IL-17A、TGF-β1在3个时间点的表达量均升高(P<0.05),IL-17A在3 d达峰,IL-6在7 d达峰,IL-10、TGF-β1则为逐渐升高趋势(P<0.05)。结论Th17、Treg及其炎症因子的演变可能参与了MsPGN的发病与转归。

Objective To investigate the evolution characteristics of helper T cell 17(Th17),regulatory T cell(Treg)and inflammatory factors in peripheral blood of rat model of anti-Thy1.1 mesangial proliferative glomerulonephritis(MsPGN)Methods Clean male SD rats were randomly divided into normal group(n=18)and MsPGN group(n=18).MsPGN model,induced by single injection of 1.29 mg/kg anti-rat CD90(Thy1.1)through tail vein at 3,7 and 14 days,were randomly selected for renal tissue HE,PAS staining and IL-17A,Foxp3 immunohistochemical analysis;24 h urine protein,blood urea and creatinine contents were determined.The composition of Th17 Treg in peripheral blood was detected by flow cytometry.Moreover,IL-17A,IL-6,IL-10 and TGF-β1 in serum were detected by ELISA.Results Compared with normal group,the proliferation of mesangial cells and mesangial matrix in glomerular mesangial region of renal cortex were observed by HE and PAS staining in MsPGN rats,which peaked at 7 days;Immunohistochemical results showed that IL-17A was significantly increased in glomerular mesangial region of renal cortex and reached the peak at 3 d(P<0.05);meanwhile,the expression of Foxp3 decreased at 3 d and then increased and reached the peak at 7 d(P<0.05).The expression of urinary protein and blood urea of 24 h increased at three time points and reached the peak at 3 d(P<0.05).And there was no significant difference in serum creatinine.Flow flow results showed that Th17 increased at different time points and reached the peak at 3 d,while Treg was lower than the control at different time points with an increasing trend(P<0.05).ELISA data showed that the expression of all of IL-6,IL-17A and TGF-β1 increased at three time points(P<0.05),in which the expression of IL-17A and IL-6 reached the peaked at 3 and 7 d respectively,while the expression of IL-10 and TGF-β1 increased gradually(P<0.05).Conclusion The evolution of Th17 Treg and its inflammatory cytokines may be involved in the pathogenesis and outcome of MsPGN.