血管紧张素Ⅱ及氯沙坦对糖尿病大鼠心肌成纤维细胞α_1(Ⅰ)前胶原mRNA和蛋白表达的影响  

Effects of ATⅡ and ATⅡ receptor subtype antagonist on expression of pro α_1 (Ⅰ) collagen mRNA and protein of cardiac fibroblasts in diabetic rats

作　　者：张琳[1] 周丽诺[1] 吴烯[1] 沈稚舟[1] 朱禧星[1] 

机构地区：[1]复旦大学附属华山医院内分泌科复旦大学内分泌糖尿病研究所,上海200040

出　　处：《中华糖尿病杂志（1006-6187）》2005年第2期134-136,共3页

基　　金：上海市科委自然科学基金资助项目(002B14061)

摘　　要：目的　探讨血管紧张素Ⅱ(ATⅡ)及其1 型受体阻断剂氯沙坦对糖尿病动物模型心肌成纤维细胞α1(Ⅰ)前胶原mRNA和蛋白表达的作用。　方法　用链脲佐菌素制备糖尿病大鼠模型,饲养12周后分离心肌成纤维细胞进行原代培养。分别将一定浓度的AT Ⅱ(10-9~10-7 mmol/L),和ATⅡ10-7 mmol/L加上不同浓度ATⅡ受体阻滞剂氯沙坦(10-7~10-5 mmol/L)加入细胞培养液中刺激48 h,分别采用逆转录聚合酶链反应(RT-PCR)及Western 印迹方法检测大鼠心肌成纤维细胞α1(Ⅰ)前胶原的mRNA及蛋白表达。　结果　体外成功地培养糖尿病心肌成纤维细胞,并经免疫组化染色结果证实。经ATⅡ刺激48 h后,α1(Ⅰ)前胶原mRNA和蛋白表达量明显增高,增高程度与ATⅡ浓度呈正比;加入氯沙坦后,前胶原mRNA和蛋白表达量较单用ATⅡ组下降,也呈浓度依赖性。　结论　ATⅡ可促进糖尿病大鼠心肌成纤维细胞的胶原合成,可能在糖尿病大鼠心肌纤维化倾向中起重要作用,该作用可能主要通过ATⅡ1型受体介导完成。Objective To examine the effects of ATⅡ and ATⅡ receptor subtype antagonist on expression of pro α_1(Ⅰ) collagen mRNA and protein of cardiac fibroblasts in diabetic rat. (Methods) Myocardial fibroblasts were isolated for primary culture from three-month-old, male, (streptozotocin-)induced diabetic rats and control Sprague-Dawley rats. The fibroblasts were identified with (immuno-)histochemical method. Myocardial fibroblasts were treated with different concentrations of ATⅡ (10^(-9)～10^(-7) mmol/L) or ATⅡ (10^(-7) mmol/L) plus losartan (10^(-7)～10^(-5) mmol/L) for 48 h (separately.) The expression of pro α_1 (Ⅰ) collagen was examined by semi- quantitative RT-PCR and Western blot analyses in different concentration of these agents. Results In diabetic group, the (expression) of pro α_1 (Ⅰ) collagen mRNA and protein of these cells rose gradually according to the (concentrations) of ATⅡ. But this effect could be blocked partly or completely when ATⅡ (10^(-7)) were plused with serial concentrations of losartan. These results suggested that ATⅡ can enhance and losartan can block the expression of pro α_1(Ⅰ) collagen mRNA and protein in myocardial fibroblasts. Conclusions ATⅡ might take a great part in cardiac fibrosis and cardiomyopathy in diabetic heart. Such function may be blocked by ATⅡ receptor antagonist.

关 键 词：血管紧张素Ⅱ 氯沙坦 糖尿病 大鼠 心肌 纤维细胞α1 胶原MRNA 蛋白表达 逆转录聚合酶链反应 RT-PCR 

分 类 号：R587.2[医药卫生—内分泌]