过刊浏览

川芎嗪抑制血管紧张素Ⅱ诱导的平滑肌细胞NF--#kappa#B激活和骨形成蛋白--2表达降低

任新瑜, 阮秋蓉, 朱大和, 朱敏, 瞿智玲, 路军

华中科技大学同济医学院病理系.湖北,武汉 430030

摘要

该文旨在观察血管紧张素Ⅱ（angiotensinⅡ，AngⅡ）对血管平滑肌细胞核转录因子--#kappa#B（nuclear factor--#kappa#B，NF--#kappa#B）的活性及骨形成蛋白--2（bone morphogenetic protein--2，BMP--2）表达的影响，以探讨AngⅡ参与动脉粥样硬化的机制，并探讨川芎嗪是否能抑制AngⅡ的促动脉粥样硬化作用。采用Western blot、免疫组化和原位杂交等方法分别检测AngⅡ刺激和川芎嗪干预后NF--#kappa#B活性、BMP--2蛋白和mRNA表达的变化。结果显示：（1）AngⅡ刺激激活NF--#kappa#B。AngⅡ刺激15min即有NF--#kappa#B p65核转移，30min达高峰（{sl P}<0.01），1h后减退。川芎嗪抑制AngⅡ诱导的NF--#kappa#B激活，与AngⅡ组比较，川芎嗪+AngⅡ组NF--#kappa#B活性显著降低（{sl P}<0.01）。（2）AngⅡ刺激6h时BMP--2表达增强（{sl P}<0.05），12h时减弱（{sl P}<0.01），24h时更弱（{sl P}<0.01）。川芎嗪+AngⅡ组中，川芎嗪干预6h时BMP--2表达亦增强，12与24h时保持正常水平。（3）川芎嗪对正常细胞的NF--#kappa#B活性和BMP--2表达无影响。以上结果表明，AngⅡ刺激后激活NF--#kappa#B并最终使生长抑制因子BMP--2表达下降，这可能是其参与动脉粥样硬化发生的机制之一。BMP--2一过性增高可能不依赖NF--#kappa#B通路的激活。川芎嗪可抑制AngⅡ诱导的NF--#kappa#B激活与BMP--2表达降低，提示它在抗动脉粥样硬化形成中起重要作用。

关键词： 骨形成蛋白-2; 核转录因子-#kappa#B; 血管紧张素Ⅱ; 动脉粥样硬化; 川芎嗪

Tetramethylpyrazine inhibits agiontensin H--induced nudear factor--#kappa#B activation and bone morphogenetic protein--2 downregulation in rat vascular smooth muscle cells

Ren Xinyu, Ruan Qiurong, Zhu Dahe, Zhu Min, Qu Zhiling, Lu Jun

Department of Pathology, Tongji Medical College of Huazhong University of Science and Technology.Wuhan 430030,Hubei

Abstract

Tetramethylpyrazine （TMP）, an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin Ⅱ （Ang Ⅱ） is highly implicated in the proliferation of vascular smooth muscle cells （VSMCs）, resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang H remain to be defined. The present study was aimed to study the effect of TMP on Ang Ⅱ-induced VSMC proliferation through detection of nuclear factor-#kappa#B （NF-#kappa#B） activity and bone morphogenetic protein-2 （BMP-2） expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang Ⅱ group, Ang Ⅱ + TMP group and TMP group. Cells in each group were harvested at different time points （15, 30 and 60 min for detection of NF-#kappa#B activity; 6, 12 and 24 h for measurement of BMP-2 expression）. NF-#kappa#B activation was identified as nuclear staining by irnmtmohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that： （1） Ang Ⅱ stimulated the activation of NF-#kappa#B. Translocation of NF-#kappa#B p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min （P〈0.01） and declined after 1 h. （2） TMP inhibited Ang H-induced NF-#kappa#B activation （P〈 0.01）. （3） Ang H increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h （P〈0.01）. （4） BMP-2 expression was also kept at high level at 6 h in Ang Ⅱ + TMP group but maintained at the normal level at 12 and 24 h. （5） There was no significant difference in NF-#kappa#B activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang Ⅱ-induced VSMC proliferation through repression of NF-#kappa#B activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-#kappa#B pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.

Key words: bone morphogenetic protein-2;nuclear factor-#kappa#B;angiotensinⅡ;Atherosclerosis;Tetramethylpyrazine

收稿日期：　　录用日期：

通讯作者：　　E-mail:

引用本文：

任新瑜, 阮秋蓉, 朱大和, 朱敏, 瞿智玲, 路军. 川芎嗪抑制血管紧张素Ⅱ诱导的平滑肌细胞NF--#kappa#B激活和骨形成蛋白--2表达降低[J]. 生理学报 2007; 59 (3): 339-344.

Ren Xinyu, Ruan Qiurong, Zhu Dahe, Zhu Min, Qu Zhiling, Lu Jun. Tetramethylpyrazine inhibits agiontensin H--induced nudear factor--#kappa#B activation and bone morphogenetic protein--2 downregulation in rat vascular smooth muscle cells. Acta Physiol Sin 2007; 59 (3): 339-344 (in Chinese with English abstract).