雷帕霉素抑制大鼠单侧输尿管梗阻后肾脏结缔组织生长因子的表达

目的研究结缔组织生长因子（CTGF）在肾脏间质纤维化发病机制中的意义,探讨雷帕霉素对其的调节作用。方法建立大鼠单侧输尿管梗阻（UUO）动物模型,采用半定量逆转录聚合酶链反应（RT-PCR）和Western印迹动态观察梗阻侧肾脏及假手术组结缔组织生长因子mRNA和蛋白的表达,并于第7、14天观察雷帕霉素对CTGF和纤维连接蛋白（FN）表达的作用。结果模型组及雷帕霉素组梗阻侧肾脏CTGF均明显增加（均P〈0．01）,雷帕霉素组比模型组的CTGF mRNA在第7天表达减少（P〈0．05）,CTGF蛋白在第7天（P〈0．05）、14天（P〈0．01）表达下调并且纤维连接蛋白的表达在第7天也下调（P〈0．05）。结论输尿管梗阻后肾脏CTGF mRNA和蛋白表达明显增加,在UUO早期雷帕霉素对CTGF、FN有一定的抑制作用,从而改善肾间质纤维化。

Expression of connective tissue growth factor in renal interstitial fibrosis after ureteral obstruction and effects of rapamycin thereupon: experiment with rats

OBJECTIVE: Is a common feature of progressive renal diseases regardless of the initiating insult To clarify the role of connective tissue growth factor (CTGF) in after (UUO) in renal interstitial fibrosis and effects of rapamycin (RAP) thereupon. METHODS: Eighteen Sprague-Dawley rats were randomly divided into 3 equal groups: unilateral ureteral obstruction (UUO) model group, undergoing ligation of the left ureter; RAP treatment group, undergoing ligation of the left ureter and intraperitoneal injection of RAP 0.04 mg.kg(-1).d(-1); and sham operation group. The right kidneys were taken out 7 and 14 days after the operation respectively to undergo renal pathological examination by Masson staining. Semi-quantitative RT-PCR was used to detect the mRNA expression of CTGF. Western blotting was performed to examine the protein expression of CTGF and fibronectin (FN). RESULTS: In comparison with the sham operation group, renal interstitial fibrosis was significant more expression in the 2 UUO groups, especially the UUO model group (P < 0.01). Seven and 14 days after the operation the levels of CTGF mRNA expression of the UUO model and RAP treatment groups (both P < 0.01), and the level of CTGF mRNA expression of the RAP treatment group was significantly lower than that of the UUO model group (P < 0.01), however, there was no significant difference in the level of CTGF mRNA expression between the 2 UUO groups 14 days after the operation. Seven and 14 days after the operation the levels of CTGF protein expression of the UUO model and RAP treatment groups were both significantly higher than that of the sham operation group (both P < 0.01), and the levels of CTGF protein expression of the RAP treatment group were significantly lower than that of the UUO model group (P < 0.05 and P < 0.01). The levels of FN expression 7 and 14 days after the operation of the 2 UUO groups were both significantly higher than that of the sham operation group (both P < 0.01). and the level of FN expression 7 days after of the RAP treatment group was significantly lower than that of the UUO model group (P < 0.01), however, there was no significant difference in the level of FN expression between the 2 UUO groups 14 days after the operation. CONCLUSION: The expression of CTGF mRNA and that of CTGF protein increase after UUO. Rapamycin play a protective role in the kidney by downregulating the CTGF expression and alleviating the renal interstitial fibrosis following UUO.