【摘要】 目的 :观察低氧对结缔组织生长因子 (CTGF)表达的影响 ,探讨低氧致肾间质纤维化的机制。方法 :单侧输尿管结扎 (UUO) 9d大鼠动物模型 ,用RT -PCR方法检测肾组织中低氧标记分子 -低氧诱导因子 (HIF - 1α)的mRNA水平 ,免疫组化方法观察假手术组及模型组肾组织中HIF - 1α和CTGF的表达及部位 ,Western蛋白印迹技术检测肾组织CTGF的蛋白水平。体外实验 ,正常大鼠肾间质成纤维细胞 (NRK - 4 9F)分别置于低氧 (1%O2 )和正常氧条件下培养 6h ,应用RT -PCR和Western蛋白印迹检测CTGFmRNA和蛋白表达水平。结果 :对照组肾组织未能检测到HIF - 1αmRNA和HIF - 1α蛋白表达 ;模型组肾组织有高水平HIF - 1αmRNA ,并出现HIF - 1α蛋白表达 ,主要分布在小管 -间质细胞 ;CTGF蛋白与HIF - 1α蛋白表达部位及程度一致 ;低氧 (1%O2 )培养刺激NRK - 4 9F细胞表达CT GFmRNA和蛋白。结论 :低氧刺激的CTGF的表达增加与肾间质纤维化的发生有关。更多还原

【Abstract】 AIM: To study the expression of connective tissue growth factor (CTGF) induced by hypoxia, and the role and mechanism of hypoxia on promoting renal interstitial fibrosis. METHODS: Renal interstitial fibrosis was induced by unilateral ureteral obstruction (UUO) in rat animal model for 9 days as in vivo studies; marker of hypoxia-HIF-1α mRNA and protein, the expression of CTGF in the obstructed kidneys were assessed by RT-PCR, immunohistochemistry and Western blotting respectively. In vitro , normal rat kidney interstitial fibroblast cells (NRK-49F) were exposed to hypoxia (1%O 2) for up to 6 hours, hypoxia was confirmed by detecting the expression of HIF-1α protein in cells, cellular level of CTGF mRNA and protein were assessed by RT-PCR and Western blotting respectively. RESULTS: Neither HIF-1α mRNA nor HIF-1α protein was expressed in the kidney from sham-operated group of rats. High level of HIF-1α mRNA were occurred, and strongly HIF-1α positive immunostaining were seen in the tubular and interstitial cells in kidney from UUO rats. Expression and location of CTGF protein were paralleled and relevant with the expression of HIF-1α protein in kidney of UUO rats. In cultured NRK-49F cell line, subjected to hypoxia even for 6 hours stimulated the expression of CTGF mRNA and protein. CONCLUSION: Our results indicated that hypoxia could stimulate the expression of CTGF mRNA and protein in kidney from UUO rats, which may in turn contribute to renal interstitial fibrosis.更多还原