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探讨白细胞介素17(IL-17)在肾间质纤维化发生发展中的作用。采用的体内实验为,以单侧输尿管梗阻(unilateral ureteric obstruction,UUO)启动纤维化病理过程。将C57BL/6小鼠分为假手术组(Sham)和单侧输尿管梗阻(unilateral ureteric obstruction,UUO)组,分别于术后第1、3、7天处死,留取手术侧肾组织。采用HE、PAS、PASM、Masson染色评价肾间质组织病理变化。以免疫组化、实时荧光定量PCR(qRT-PCR)法检测α平滑肌肌动蛋白(α-SMA)表达水平;酶联免疫吸附试验(ELISA)检测IL-17在肾组织中的表达情况。体外实验为,分离培养C57BL/6小鼠肾成纤维细胞,转化生长因子β1(TGF-β1)刺激使其转化为肌成纤维细胞,继而用蛋白免疫印迹法(western blotting)检测IL-17的刺激下α-SMA表达水平。结果显示,组织病理学检查显示随着梗阻时间延长,炎性细胞浸润明显,肾小管萎缩,间质面积增大,Ⅰ型胶原和α-SMA增多。PCR检测显示α-SMA mRNA表达增多。但ELISA结果显示肾组织中IL-17含量呈下降趋势。Western blot结果表明肌成纤维细胞在IL-17的刺激下,α-SMA蛋白水平显著下降。实验说明,单侧输尿管梗阻手术可导致相应侧肾组织发生纤维化病变,但肾组织中IL-17含量与纤维化病变并不一致,在体外实验中IL-17可下调肌成纤维细胞表达的α-SMA。
To investigate the role of interleukin-17 (IL-17) in the development of renal interstitial fibrosis. In vivo experiments were performed to initiate fibrotic pathology with unilateral ureteric obstruction (UUO). C57BL / 6 mice were divided into sham operation group and unilateral ureteric obstruction (UUO) group, which were sacrificed on the 1st, 3rd, 7th day after surgery, respectively. The pathological changes of renal interstitium were evaluated by HE, PAS, PASM and Masson staining. The expression of α-smooth muscle actin (α-SMA) was detected by immunohistochemistry and qRT-PCR. The expression of IL-17 in renal tissues was detected by enzyme-linked immunosorbent assay (ELISA) In vitro experiments, renal fibroblasts of C57BL / 6 mice were isolated and cultured and transformed into myofibroblasts stimulated by transforming growth factor-β1 (TGF-β1). Then IL-17 was detected by western blotting Stimulation of α-SMA expression levels. The results showed that histopathological examination showed that as the obstruction time prolonged, infiltration of inflammatory cells was obvious, tubular atrophy, interstitial area increased, type I collagen and α-SMA increased. PCR detection showed that α-SMA mRNA expression increased. However, the results of ELISA showed that the content of IL-17 in renal tissue showed a decreasing trend. Western blot results showed that IL-17 stimulated by myofibroblasts, α-SMA protein levels decreased significantly. Experiments show that unilateral ureteral obstruction surgery can lead to fibrosis of the corresponding side of renal tissue, but the content of IL-17 in renal tissue and fibrosis is not consistent, in vitro experiments IL-17 can be down regulated myofibroblasts α -SMA.