血管紧张素对增生性瘢痕成纤维细胞纤维连接蛋白合成的影响

Effects of angiotensin II on the expression of fibronectin of fibroblast from hypertrophic scar

目的:探讨血管紧张素Ⅱ(AngⅡ)及其I型、Ⅱ型受体阻断剂和钙调神经磷酸激酶(CaN)的阻滞剂环胞素A(CsA)对增生性瘢痕来源的成纤维细胞纤维连接蛋白mRNA和蛋白表达的作用。方法:体外分离培养增生性瘢痕成纤维细胞,分别将一定浓度的AngⅡ(10-9～10-5mmol/L),和AngⅡ10-6mmol/L加上不同阻滞剂losartan、PD123319、环胞素A(浓度均为10-5mmol/L)加入细胞培养液中刺激48h,分别采用逆转录聚合酶链反应(RT-PCR)及Westernblot印迹方法检测增生性瘢痕成纤维细胞纤维连接蛋白(FN)的mRNA及蛋白表达。结果:体外成功地培养增生性瘢痕成纤维细胞。经AngⅡ刺激48h后,FNmRNA和蛋白表达量明显增高,增高程度与AngⅡ浓度呈正比;加入losaartan和环胞素A后,FN原mRNA和蛋白表达量较单用AngⅡ组下降,而加入PD123319后,FNmRNA和蛋白表达量较单用AngⅡ组无明显改变。结论:AngⅡ可促增生性瘢痕成纤维细胞的FN合成,可能在增生性瘢痕的发展过程中起重要作用,该作用主要通过AngⅡ的I型受体介导完成,该作用可能与CaN信号通路有关。

Objective To investigate the effect of angiotensin Ⅱ（Ang Ⅱ） and Ang Ⅱ receptor subtype antagonist and CaN antagonist （CsA） on the expression of fibronectin of fibroblast from hypertrophic scar. Methods Fibroblasts were freshly isolated from hypertrophic scars and were cultured.The fibroblasts were treated with different concentration of An g ll （10^-9-10^-5mmol/L） or Angll （10^-6mmol/L） plus Iosartan （10^-5mmol/L）. PD 123319（10^-5mmol/L）. CsA （10^-5mmol/L） for 48h separately.The expression of fibronectin was examined by semi-quantitative RT-PCR and Western blot. Results The expression of fibronectin mRNA and protein of these cells rose gradually according to the concentration of Angll. But this effect could be blocked partly or completely by Iosartan and CsA,while PD12319 has no effect. Conclusion Angll might take a great part in the development of hypertrophic scar via AT1R. CaN signal pathway might involve in this process.