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。
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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2005年第3期432-435,共4页
Chinese Journal of Pathophysiology
基金
国家自然科学基金 (No .30 2 70 6 12 )
教育部教育振兴行动计划 (985工程 )专项科研基金资助
关键词
低氧
结缔组织生长因子
低氧诱导因子
纤维化
肾
Anoxia
Connective tissue growth factor
Hypoxia inducible factor
Fibrosis
Kidney
