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目的:研究香青兰总黄酮对血管紧张素Ⅱ(AngⅡ)诱导的大鼠主动脉血管平滑肌细胞(VSMC)黏附分子及基质金属蛋白酶(MMPs)表达的影响。方法:采用贴壁法培养大鼠胸主动脉平滑肌细胞,以AngⅡ为诱导剂,建立VSMC细胞增殖的模型,分别应用10-7 mol·L~(-1) AngⅡ以及AngⅡ+不同浓度香青兰总黄酮组(25,50,100μg·mL~(-1))作用24 h,并设空白对照组进行比较。采用免疫组化法检测细胞中细胞间黏附分子(ICAM-1)、血管间黏附分子(VCAM-1)的表达水平。RT-PCR方法检测细胞MMP-2、MMP-9的mRNA表达水平。结果:与对照组比较,AngⅡ组能显著刺激大鼠VSMC细胞内ICAM-1、VCAM-1、MMP-2、MMP-9的表达,香青兰总黄酮不同剂量组联合AngⅡ可在一定程度上抑制AngⅡ诱导的VSMC细胞内ICAM-1、VCAM-1、MMP-2、MMP-9的表达,且呈现一定的剂量依赖关系趋势。结论:香青兰总黄酮具有抑制AngⅡ诱导VSMC黏附分子及基质金属蛋白酶表达的作用。
Objective: To study the effects of total flavonoids from Herba Cymbidium on angiotensin Ⅱ (Ang Ⅱ) - induced adhesion molecules and matrix metalloproteinases (MMPs) in rat aortic vascular smooth muscle cells (VSMC). Methods: Rat thoracic aortic smooth muscle cells (VSMCs) were cultured by adherent method. The proliferation of VSMCs was induced by AngⅡ. Angiotensin Ⅱ (AngⅡ) and AngⅡ (10 -7 mol·L -1) Total flavonoids group (25,50,100 μg · mL -1) for 24 h, and set the blank control group for comparison. Immunohistochemistry was used to detect the expression of intercellular adhesion molecule (ICAM-1) and intercellular adhesion molecule (VCAM-1). The mRNA expression of MMP-2 and MMP-9 was detected by RT-PCR. Results: Compared with the control group, the AngⅡ group could significantly stimulate the expression of ICAM-1, VCAM-1, MMP-2 and MMP-9 in VSMCs of rats, Inhibited the expression of ICAM-1, VCAM-1, MMP-2 and MMP-9 in AngⅡ-induced VSMC cells in a dose-dependent manner. Conclusion: The total flavonoids of Cymbidium can inhibit the expression of adhesion molecules and matrix metalloproteinases induced by Ang Ⅱ.