摘要
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.
作者
张音
陆平
盛净
Zhang Yin;Lu Ping;Sheng Jing(Department of Geriatrics,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University,School of Medicine,Shanghai,200011,P.R.China)
出处
《老年医学与保健》
CAS
2018年第4期376-380,396,共6页
Geriatrics & Health Care
基金
国家自然基金青年科学项目(81300092)
关键词
受体
血小板源生长因子
基质金属蛋白酶
肌细胞
平滑肌
receptors
platelet-derived growth factor (PDGF)
matrix metalloproteinases (MMP)
myocytes
smooth muscle