[N-乙酰丝氨酰-天冬氨酰-赖氨酰-脯氨酸对血管紧张素II诱导的MRC-5人胚肺成纤维细胞肌成纤维细胞分化的抑制作用]

[Inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts inuced by Ang II].

作者信息

Li Shifeng, Du Shipu, Xue Xinxin, Xu Dingjie, Xu Hong, Sun Yue, Deng Haijing, Yang Yi, Wei Zhongqiu, Tian Jingrui, Yang Fang

机构信息

Hebei United University, Tangshan 063000, China.

E-mail:

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2014 Nov;32(11):801-5.

PMID:25579022

Abstract

OBJECTIVE

To explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang) II.

METHODS

The study was divided into 2 step: (1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang II and at different time point by 100 nmol/L Ang II. Then the expression of collagen type I and α-smooth muscle actin (α-SMA) were mesaured by western blot. (2) MRC-5 human fetal lung fibroblasts were divided into 4 group: (1) control, (2) Ang II, (3) Ang II+Ac-SDKP, (4) Ang II+8-Me-cAMP (a specific activator of Epac). The α-SMA expression was observed by immnocytochemical stain. The protein expression of collagen type I, α-SMA, serum response factor (SRF), myocardin-related transcription factor (MRTF)-A, exchange protein directly activated by cAMP (Epac) 1, 2 were measured by Westen blot.

RESULTS

Myofibroblast differentiation could be induced by Ang II from MRC-5 cells with a dose- and time-dependent manner. The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang II and accompanied with collagen I and α-SMA increased. Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang II, and promoted the expression of Epac1.

CONCLUSION

Ac-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang II via Epac1 activating.

摘要

目的

探讨N - 乙酰 - 丝氨酰 - 天冬氨酰 - 赖氨酰 - 脯氨酸（Ac - SDKP）对血管紧张素（Ang）II诱导的MRC - 5人胚肺成纤维细胞向肌成纤维细胞分化的抑制作用。

方法

本研究分为两步：（1）用不同剂量的Ang II及100 nmol/L Ang II在不同时间点诱导MRC - 5人胚肺成纤维细胞48小时。然后通过蛋白质免疫印迹法检测I型胶原蛋白和α - 平滑肌肌动蛋白（α - SMA）的表达。（2）将MRC - 5人胚肺成纤维细胞分为4组：（1）对照组，（2）Ang II组，（3）Ang II + Ac - SDKP组，（4）Ang II + 8 - 甲基 - cAMP（Epac的特异性激活剂）组。通过免疫细胞化学染色观察α - SMA的表达。通过蛋白质免疫印迹法检测I型胶原蛋白、α - SMA、血清反应因子（SRF）、心肌相关转录因子（MRTF） - A、cAMP直接激活的交换蛋白（Epac）1、2的蛋白表达。

结果

Ang II可诱导MRC - 5细胞向肌成纤维细胞分化，呈剂量和时间依赖性。在Ang II诱导的MRC - 5细胞中观察到SRF和MRTF - A上调，并伴有I型胶原蛋白和α - SMA增加。用8 - 甲基 - cAMP或Ac - SDKP预处理可减弱Ang II诱导的所有这些变化，并促进Epac1的表达。