CAJ | 학술논문

本文旨在观察血管紧张素Ⅱ(angiotensin Ⅱ,Ang Ⅱ)对血管平滑肌细胞核转录因子-κB(nuclear factor-κB,NF-κB)的活性及骨形成蛋白-2 (bone morphogenetic protein-2, BMP-2)表达的影响,以探讨Ang Ⅱ参与动脉粥样硬化的机制,并探讨川芎嗪是否能抑制Ang Ⅱ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Ang Ⅱ刺激和川芎嗪干预后NF-κB活性、BMP-2蛋白和mRNA表达的变化.结果显示:(1) Ang Ⅱ刺激激活NF-κB.Ang Ⅱ刺激15 min即有NF-κB p65核转移,30min达高峰(P＜0.01),1h后减退.川芎嗪抑制Ang Ⅱ诱导的NF-κB激活,与Ang Ⅱ组比较,川芎嗪+Ang Ⅱ组NF-κB活性显著降低(P＜0.01).(2) Ang Ⅱ刺激6h时BMP-2表达增强(P＜0.05),12h时减弱(P＜0.01),24h时更弱(P＜0.01).川芎嗪 + Ang Ⅱ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-κB活性和BMP-2表达无影响.以上结果表明,Ang Ⅱ刺激后激活NF-κB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-κB通路的激活.川芎嗪可抑制Ang Ⅱ诱导的NF-κB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.
본문지재관찰혈관긴장소Ⅱ(angiotensin Ⅱ,Ang Ⅱ)대혈관평활기세포핵전록인자-κB(nuclear factor-κB,NF-κB)적활성급골형성단백-2 (bone morphogenetic protein-2, BMP-2)표체적영향,이탐토Ang Ⅱ삼여동맥죽양경화적궤제,병탐토천궁진시부능억제Ang Ⅱ적촉동맥죽양경화작용.채용Western blot、면역조화화원위잡교등방법분별검측Ang Ⅱ자격화천궁진간예후NF-κB활성、BMP-2단백화mRNA표체적변화.결과현시:(1) Ang Ⅱ자격격활NF-κB.Ang Ⅱ자격15 min즉유NF-κB p65핵전이,30min체고봉(P＜0.01),1h후감퇴.천궁진억제Ang Ⅱ유도적NF-κB격활,여Ang Ⅱ조비교,천궁진+Ang Ⅱ조NF-κB활성현저강저(P＜0.01).(2) Ang Ⅱ자격6h시BMP-2표체증강(P＜0.05),12h시감약(P＜0.01),24h시경약(P＜0.01).천궁진 + Ang Ⅱ조중,천궁진간예6h시BMP-2표체역증강,12여24h시보지정상수평.(3) 천궁진대정상세포적NF-κB활성화BMP-2표체무영향.이상결과표명,Ang Ⅱ자격후격활NF-κB병최종사생장억제인자BMP-2표체하강,저가능시기삼여동맥죽양경화발생적궤제지일.BMP-2일과성증고가능불의뢰NF-κB통로적격활.천궁진가억제Ang Ⅱ유도적NF-κB격활여BMP-2표체강저,제시타재항동맥죽양경화형성중기중요작용.
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 Ⅱ 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-κB (NF-κB) 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-κB activity;6,12 and 24 h for measurement of BMP-2 expression). NF-κB activation was identified as nuclear staining by immunohistochemistry. 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-κ B. Translocation of NF-κB 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 Ⅱ-induced NF-κB activation (P＜0.01).(3) Ang Ⅱ 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-κB 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-κB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-κB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.