益气活血化瘀方药对血管胶原转换及相关基因表达的影响

Effect of Yiqi Huoxue Huayu Recipe on Vascular Collagen Turnover and Relevant Gene Expression

目的:观察益气活血化瘀中药对大鼠血管细胞外基质(ECM)重塑的影响并探讨其分子机制。方法:采用3H-TdR和3H-Pro掺入实验和血管壁羟脯氨酸含量测定检查血管平滑肌细胞(VSMC)增殖活性及胶原合成与降解程度;应用Northern印迹和Ⅳ型胶原酶(MMP-2)酶图分析检测MMP-2和骨桥蛋白表达的变化。结果:益气活血化瘀中药可显著抑制bFGF对VSMC的促胶原合成作用,降低血管内皮剥脱大鼠血管壁胶原转换速率,下调MMP-2和骨桥蛋白在体外培养的VSMC及血管壁中的表达水平,抑制VSMC增殖。中药血清去蛋白后对VSMC的影响不发生明显变化。结论:益气活血化瘀中药通过调节MMP-2和骨桥蛋白基因表达及降低胶原转换速率可抑制和(或)减缓ECM重塑。

Objective: To explore the effect of Yiqi Huoxue recipe (YHR) , a Chinese herbal medicine for supplementing Qi, activating blood circulation to remove stasis, on vascular extracellular matrix (ECM) remodeling and its molecular mechanism. Methods: Vascular smooth muscle cell (VSMC) proliferation activity and collagen turnover rate were detected by 3 H-TdR test and hydroxyproline amount determined by 3 H-Pro incorporation. Expression activity of MMP-2 and osteopontin genes was detected by Northern blotting and MMP-2 zymography analysis. Results: YHR could markedly inhibit VSMC collagen synthesis stimulated by blasic fibroblast growth factor (bFGF) and lower the collagen turnover rate induced by vascular de-endothelialization. The expression level of MMP-2 and osteopontin genes was down-regulated by YHR in cultured VSMC and vascular wall with endothelial injury, and VSMC proliferation was inhibited by the serum obtained from YHR treated rats. Removing protein from the drug serum made no change on the effect of YHR to VSMC. Conclusion: YHR could inhibit and/or retard ECM remodeling through regulating the expression of MMP-2 and osteopontin genes and lowering the collagen turnover rate.