桔梗皂甙D对PDGF-BB诱导的血管平滑肌细胞迁移的影响

Effect of Platycodin D on PDGF-BB-induced Vascular Smooth Muscle Cell Migration

目的探讨桔梗皂甙D对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)迁移的影响。方法体外培养大鼠胸主动脉VSMCs(A7r5细胞株),通过划痕实验和Transwell小室评估桔梗皂甙D以及MAPKs抑制剂(U0126/SB203580/SP600125)对血小板源性生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)诱导的VSMCs迁移和侵袭的影响。明胶酶谱法检测MMP-2和MMP-9在PDGF-BB诱导的VSMCs中的蛋白水解活性;RT-qPCR检测MMP-2和MMP-9的mRNA表达水平。各组细胞反应48h。结果桔梗皂甙D在4μg/mL时即能抑制PDGF-BB刺激VSMCs的迁移及侵袭能力,抑制MMP-9的蛋白水解活性,下调其mRNA表达(P<0.05)。MAPKs抑制剂20μM能抑制PDGFBB刺激的VSMCs迁移及侵袭及MMP-9的mRNA表达(P<0.01),与MAPKs抑制剂20μM+桔梗皂甙D4μg/mL组比较,2组抑制PDGF-BB促细胞迁移及侵袭的能力、下调MMP-9mRNA表达水平差异无统计学意义(P>0.05)。结论桔梗皂甙D抑制了MMP-9的活性并下调其mRNA表达,对PDGF-BB诱导的VSMCs迁移与侵袭有抑制作用,其机制可能与部分抑制MAPKs的活性有关。

Objective To investigate the effect of platycodin D（PD） on vascular smooth muscle cells（VSMCs）migration. Methods Rat thoracic aortic vascular smooth muscle cells（A7 r5 cell line） were cultured in vitro; the effects of platycodin D and MAPKs inhibitor（U0126/SB203580/SP600125） on the migration and invasion of VSMCs induced by plateletderived growth factor-BB（PDGF-BB） were evaluated by scratch test and Transwell chamber; the proteolytic activity of MMP-9 and MMP-2 in PDGF-BB-induced VSMCs was detected by gelatin zymography and the mRNA expression of matrix metalloproteinases 2（MMP-2） and matrix metalloproteinases 9（MMP-9） by reverse transcription quantitative polymerase chain reaction（RT-qPCR）; all the cells reacted for 48 hours. Results Platycodin D at the concentration of 4 μg/mL significantly inhibited the migration and invasion of VSMCs stimulated by PDGF-BB and the proteolytic activity of MMP-9, and, down-regulated its mRNA expression（P0.05）; MAPKs inhibitor at 20μM significantly inhibited the migration and invasion of VSMCs stimulated by PDGF-BB and the expression of MMP-9 mRNA（P0.01）; there was no significant difference in inhibiting the ability of PDGF-BB to promote cell migration and invasion and in down-regulating the mRNA expression of MMP-9 between MAPKs inhibitor at 20 μM and MAPKs inhibitor 20 μM ＋ PD 4 μg/mL（P0.05）. Conclusions Platycodin D can inhibit the activity of MMP-9 and down-regulate its mRNA expression, effective in inhibiting the migration and invasion of V SMCs induced by PDGFBB, which may be partly related to the inhibition of MAPKs activity.