解毒消癥饮对血管生成因子A诱导人脐静脉内皮细胞的体外血管生成抑制作用

Inhibition of Jiedu Xiaozheng Yin on the angiogenesis of human umbilical vein endothelial cells induced by vascular endothelial growth factor in vitro

目的:探讨解毒消癥饮(JXY)对血管生成因子A(VEGFA)诱导的人脐静脉内皮细胞(HUVECs)凋亡、迁移以及血管生成的体外影响。方法:HUVECs分为对照组、诱导组(VEGFA 10ng/mL)、JXY低剂量组(VEGFA10ng/L+JXY0.05mg/mL)、JXY中剂量组(VEGFA10ng/mL+JXY0.1mg/mL)、JXY高剂量组(VEGFA10ng/mL+JXY 0.2mg/mL),通过MTT、Hoechst、划痕实验,管腔形成实验、Transwell以及Western blot分别检测细胞活力、凋亡、修复能力、血管生成、细胞迁移和VEGFR2、MMP2、MMP9蛋白表达。结果:与诱导组比较,JXY以剂量依赖性地抑制VEGFA诱导的细胞增殖、修复能力,迁移以及体外管腔的形成能力(P<0.01),促进凋亡(P<0.05,P<0.01)。进一步研究表明JXY剂量依赖性降低VEGFR2、MMP2、MMP9蛋白表达量。结论:JXY显著抑制VEGFA诱导的HUVECs细胞体外血管生成,与其降低VEGFR2、MMP2、MMP9表达直接相关。

Objective: To investigate the effect of Jiedu Xiaozheng Yin(JXY) on apoptosis, migration and angiogenesis of human umbilical vein endothelial cells(HUVECs) induced by angiogenic factor A(VEGFA). Methods: HUVEC cells were divided into 5 groups: Control group, induction group(10 ng/mL VEGFA), JXY low dose group(VEGFA 10 ng/mL+JXY 0.05 mg/mL), JXY middle dose group(VEGFA 10 ng/mL+JXY 0.1 mg/mL), and JXY high dose group(VEGFA 10 ng/mL+ JXY 0.2 mg/mL). MTT and Hoechst were used to detect cell viability and apoptosis. And cell repair ability and angiogenesis of HUVECs were assayed by wound healing and tube formation. Cell migration was detected by transwell and some key proteins level such as VEGFR2, MMP2 and MMP9 were detected by Western blot assay. Results: Compared with the induction group, JXY inhibited VEGFA-induced HUVEC cells proliferation and reduced HUVEC cells repair ability and migration ability, lowed the tube formation ability of HUVECs(P<0.01), promoted HUVEC cells apoptosis(P<0.05, P<0.01). Furthermore, JXY downregulated the key proteins(VEGFR2, MMP2, MMP9) level in a dose-dependent manner. Conclusion: JXY significantly inhibits VEGFA-induced angiogenesis of HUVECs in vitro which potentially is due to the lowered level of VEGFR2, MMP2 and MMP9.