黄芪对血小板源生长因子诱导血管平滑肌细胞的生物学影响

Biological effects of astragalus membranaceus on vascular smooth muscle cells induced by platelet-derived growth factor

目的探讨黄芪对血小板源生长因子(platelet-derived growth factor,PDGF)诱导血管平滑肌细胞(vascular smooth muscle cells,VSMCs)生物活性的影响。方法采用噻唑蓝(methyl-thiazolyl-tetrazolium,MTT)法测定细胞增殖情况,采用流式细胞术观察细胞周期阻滞的影响,采用Transwell小室测试细胞迁移情况。采用Western blotting检测各组基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)、磷酸化细胞外调节蛋白激酶(phosphorylated extracellular regulated protein kinases 1/2,p-ERK1/2)和磷酸化蛋白激酶B(phosphorylated protein kinase B,P-AKT)的蛋白表达。结果 PDGF可促进VSMCs的增殖及迁移,并增加VSMCs的MMP-2、p-ERK1/2和p-AKT的蛋白表达。黄芪则可抑制PDGF诱导的VSMCs的增殖和迁移,诱导VSMCs细胞周期阻滞在G0/G1期且呈剂量依赖性,并降低VSMCs的MMP-2、p-ERK1/2和p-AKT蛋白的表达。黄芪注射液溶剂对照组在各组实验与PDGF-BB组相比,差异均无统计学意义(P>0.05)。结论黄芪对PDGF-BB诱导的VSMCs增殖和迁移有明显的抑制作用,其分子机制可能与抑制ERK1/2/AKT活化有关。

Objectives To explore the biological effects of astragalus membranaceus on vascular smooth muscle cells（VSMCs） induced by platelet-derived growth factor（PDGF）. Methods Methyl-thiazolyl-tetrazolium（MTT） test was used to measure the cell proliferation and flow cytometry was used to measure the cell cycle arrest. Transwell chamber assay was used to measure the cell migration. Western blotting was used to measure the protein levels of matrix metalloproteinases-2（MMP-2）, phosphorylated extracellular regulated protein kinases 1 / 2（p-ERK1 / 2） and phosphorylated protein kinase B（p-AKT）. Results Treatment with PDGF-BB significantly promoted the proliferation and migration of VSMCs and increased the protein expressions of MMP-2, p-ERK1 / 2 and p-AKT. Astragalus membranaceus significantly inhibited the proliferation and migration of VSMCs induced by PDGF-BB and decreased the protein expressions of MMP-2, p-ERK1 / 2 and p-AKT. The cell cycle of VSMCs was arrested in G0 / G1 phase in a dose-dependent manner. There was no obvious difference between astragalus solution group and PDGF-BB group（P〉0.05）. Conclusions Astragalus membranaceus can suppress the proliferation and migration of VSMCs induced by PDGF-BB. The molecular mechanism may be related to the inhibition of ERK1 / 2 / AKT activation.