四妙勇安汤对血管平滑肌细胞迁移的影响

Study on the effects of Simiao Yong An Decoction on the vascular smooth muscle cell migration

目的:研究四妙勇安汤在血管内皮细胞生长因子(VEGF)诱导条件下对血管平滑肌细胞(VSMCs)迁移的影响,并探讨其相关机制。方法:体外培养VSMCs,在VEGF诱导条件下,用四妙勇安汤冻干粉进行干预,划痕实验、Transwell检测VSMCs的迁移能力;PT-PCR检测对基质金属蛋白酶(MMP)MMP-9mRNA表达的影响;Western blot法检测对金属蛋白酶组织抑制剂(TIMP)TIMP-1、TIMP-2蛋白表达的影响。结果:①在VEGF诱导条件下,四妙勇安汤1.65×10-5、1.65×10-6、1.65×10-7g/mL可抑制VSMCs的迁移;②四妙勇安汤1.65×10-5、1.65×10-6g/mL可下调VEGF诱导的VSMCs MMP-9 mRNA的表达;③四妙勇安汤1.65×10-5、1.65×10-6、1.65×10-7g/mL均可上调VEGF诱导的VSMCs TIMP-2蛋白的表达。结论:四妙勇安汤可抑制VEGF诱导的VSMCs迁移。四妙勇安汤通过抑制VEGF诱导的VSMCs MMP-9的表达,促进VEGF诱导的VSMCs TIMP-2的表达起到抑制VEGF诱导的VSMCs迁移的作用。

Objective： To investigate the effects of Simiao Yong An Decoction on the migration of vascular smooth muscle cells （VSMCs） induced by the vascular endothelial growth factor （VEGF） and its correlated mechanisms. Methods： VSMCs were cultured. Under VEGF induced conditions, the VSMCs migration was detected using the scratch test and Transwell after intervened by Simiao Yong An Decoction. The effects on the mRNA expressions of matrix metalloproteiuase MMP-9 were detected using RT-PCR. Effects on the protein expressions of the tissue inhibitor of metalloproteinase-1 （TIMP-1） and T1MP- 2 were detected using Western blot. Results： ①Simiao Yong An Decoction （1.65×10^-5, 1.65×10^-6, 1.65×10^-7g/mL） could inhibit VEGF-induced VSMCs migration. ②Simiao Yong An Decoction （1.65×10^-5, 1.65×10^-6g/mL） could down-regulate the VEGF- induced VSMCs MMP-9 mRNA expression. ③Simiao Yong An Decoction （1.65×10^-5, 1.65×10^-6, 1.65×10^-7g/mL） up-regulated VEGF-induced VSMCs TIMP-2 protein expressions. Conclusion： Simiao Yong An Decoction could inhibit VEGF-induced VSMCs migration by inhibiting the MMP-9 mRNA expression, and increasing the TIMP-2 protein expression.