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[血管平滑肌细胞中转化生长因子-β1/信号转导分子Smad通路与细胞外信号调节激酶通路之间的相互作用]

[Interaction between TGF-beta1/Smad pathway and ERK pathway in vascular smooth muscle cells].

作者信息

Zhong Hua, He Fang, Hu Qin-hua, Wang Zhen-huan, Deng Feng-mei, Sun Zhi-ping, Li Zeng-chun

机构信息

Ministry of Education Key Laboratory of Xinjiang Endemic and Ethnic Diseases, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2010 Feb;26(1):15-8.

Abstract

OBJECTIVE

To investigate if the interaction between TGF-beta1/Smad pathway and ERK pathway in vascular smooth muscle cells exists.

METHODS

The rat arota was removed. The primary VSMC were isolated and cultured in vitro, then the VSMC were divided into four groups: (1) control group, (2) (TGF-beta1 group, (3) ERK blocking agent group, (4) TGF-beta1 + ERK blocking agent group. The expression of Smad2/3, ERK1/2 proteins, the content of phosphorylated ERK1/2 and Smad2/3 proteins were detected by Western blot, and the expression of Smad2/3 mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) .

RESULTS

(1) In contrast to control group, the content of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 group was increased (P < 0.05), that in ERK blocking agent group was decreased (P < 0.05). There was no difference between control group and TGF-beta1 + ERK blocking agent group. Compared with TGF-beta1 group, the contents of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 + ERK blocking agent group was decreased (P < 0.05). There was no difference in the expression of Smad2/3 and ERK1/2 proteins among different groups. (2) There were no differences in expression of Smad2 and Smad3 mRNA among different groups.

CONCLUSION

(1) TGF-beta1 can induce Smad2/3 proteins to be phosphorylated dependent on the activated ERK pathway. (2) ERK pathway does not effect the expression of Smad2/3 at the level of protein and mRNA.

摘要

目的

研究血管平滑肌细胞中转化生长因子β1(TGF-β1)/Smad信号通路与细胞外调节蛋白激酶(ERK)信号通路之间是否存在相互作用。

方法

取大鼠主动脉,体外分离培养血管平滑肌细胞(VSMC),将其分为4组:(1)对照组;(2)TGF-β1组;(3)ERK阻断剂组;(4)TGF-β1+ERK阻断剂组。采用蛋白质免疫印迹法检测Smad2/3、ERK1/2蛋白表达及磷酸化ERK1/2、Smad2/3蛋白含量;采用逆转录-聚合酶链反应(RT-PCR)检测Smad2/3 mRNA表达。

结果

(1)与对照组比较,TGF-β1组磷酸化Smad2/3、磷酸化ERK1/2蛋白含量增加(P<0.05),ERK阻断剂组磷酸化Smad2/3、磷酸化ERK1/2蛋白含量降低(P<0.05);对照组与TGF-β1+ERK阻断剂组比较差异无统计学意义。与TGF-β1组比较,TGF-β1+ERK阻断剂组磷酸化Smad2/3、磷酸化ERK1/2蛋白含量降低(P<0.05)。不同组间Smad2/3、ERK1/2蛋白表达差异无统计学意义。(2)不同组间Smad2、Smad3 mRNA表达差异无统计学意义。

结论

(1)TGF-β1可通过激活ERK信号通路诱导Smad2/3蛋白磷酸化。(2)ERK信号通路对Smad2/3蛋白及mRNA表达无影响。

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