Sonic hedgehog信号通路在单侧输尿管梗阻大鼠肾组织中的表达变化及意义

Expression of Sonic hedgehog pathway-related genes in kidney tissues of rats with unilateral ureteral obstruction

目的:探讨Sonic hedgehog(Shh)信号通路在单侧输尿管梗阻(UUO)大鼠肾组织中的表达变化及意义。方法:将48只SD大鼠随机分为UUO模型组(n=24)和假手术组(n=24),梗阻术3、7和14 d后取其梗阻侧肾脏组织。用HE和Masson染色检测肾间质纤维化程度,免疫组织化学染色检测Shh通路分子Shh、Ptch1、Smo、Gli1及Ⅲ型胶原的蛋白表达,酶联免疫吸附实验(ELISA)检测肾组织中TGF-β1和Shh含量,real-time RT-PCR检测TGF-β1、I和Ⅲ型胶原及Shh通路分子mRNA表达。结果:HE和Masson染色显示,梗阻侧肾组织出现明显的纤维化病变,且随时间延长而加剧。TGF-β1、I和Ⅲ型胶原含量在梗阻肾中表达明显增高(P<0.05)。同时,Shh信号通路分子Shh、Smo和Gli mRNA和蛋白在梗阻肾中表达明显升高(P<0.05),而Ptch1 mRNA和蛋白的表达下调(P<0.01),提示Shh信号被激活。相关分析表明,Shh信号起始信号Shh水平的升高与TGF-β1含量增加呈明显的相关。结论:UUO大鼠诱导肾间质纤维化发生过程中,Shh信号通路分子被激活,推测可能的机制是活化的Shh信号通路诱导TGF-β1表达和释放,导致肾间质纤维化。

AIM： To investigate the role of Sonic hedgehog （Shh） signaling pathway in renal interstitial fibro- sis in the rats with unilateral ureteral obstruction （UUO）. METHODS： Forty- eight male Sprague- Dawley rats were di- vided randomly into sham operation group and UUO model group with 24 rats each. The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fiber in the kidneys was detected with HE and Masson staining. Immunohistochemi- cal analysis was performed to evaluate the expression of Shh signaling pathway -related proteins, including Shh, Smo, Ptch1 and Gli1. The contents of TGF - β1 and Shh in the kidney tissues were determined by ELISA. Real - time RT - PCR was used to detect the mRNA expression of TGF - β1, Col Ⅰ, Col Ⅲ and Shh signaling - related genes. RESULTS ： Fibro- sis observed with HE and Masson staining was obviously increased in UUO kidneys, and aggravated as time prolonged. The contents of TGF - β1, Col Ⅰ and Col Ⅲ were also increased. In addition, the expression of Shh, Smo and Gli1 was markedly increased in obstructive kidneys, and the expression of Ptch1 was decreased （P 〈0. 01 ）, suggesting that Shh signaling was activated. The level of Shh in UUO rats was associated with the content of TGF -β1. CONCLUSION： Shh signaling is acti- vated in the progress of renal interstitial fibrosis in UUO rats, and the possible mechanism triggering the fibrogenic response is that Shh signaling promotes the expression of TGF - β1.