华良姜素对PDGF-BB诱导血管平滑肌细胞增殖和迁移的影响

Effects of jaranol on proliferation and migration of vascular smooth muscle cells induced by PDGF-BB

[目的]探讨华良姜素对PDGF-BB诱导下血管平滑肌细胞增殖和迁移的影响及可能的机制。[方法]利用25μg/L的PDGF-BB处理人主动脉血管平滑肌细胞(HA-VSMC)诱导血管平滑肌细胞增殖和迁移。实验分为正常对照组、PDGF-BB组(模型组)、PDGF-BB+华良姜素(10、20、40μmol/L)组和DMSO组。采用CCK-8实验检测HA-VSMC增殖活力,Transwell实验检测HA-VSMC迁移能力,Western blot检测自噬效应蛋白p62、LC3、雷帕霉素靶蛋白(mTOR)以及磷酸化mTOR(p-mTOR)蛋白表达水平的变化。[结果]PDGF-BB处理后,HA-VSMC增殖和迁移能力显著上调(P<0.01),华良姜素显著抑制PDGF-BB诱导的HA-VSMC的增殖和迁移,且随浓度的上升抑制能力越明显(P<0.05)。与模型组比较,40μmol/L华良姜素处理后,LC3Ⅰ以及mTOR蛋白表达水平显著降低(P<0.05);LC3Ⅱ/LC3Ⅰ、p62与p-mTOR蛋白表达水平显著升高(P<0.05),同时发现40μmol/L华良姜素与AMPK抑制剂Compound C,对PDGF-BB诱导的HA-VSMC AMPK/mTOR信号通路的影响一致。[结论]华良姜素可抑制PDGF-BB诱导的HA-VSMC的增殖和迁移,其作用机制可能是通过上调AMPK/mTOR信号通路,抑制自噬实现。

Aim To investigate the effect of jaranol on platelet derived growth factor-BB(PDGF-BB)-induced proliferation and migration of vascular smooth muscle cell(VSMC)as well as its possible mechanism.Methods Human aortic vascular smooth muscle cells(HA-VSMC)were treated with 25μg/L PDGF-BB to induce proliferation and migration.The cells were divided into normal control group(NC group),PDGF-BB group,PDGF-BB+jaranol(10,20 and 40μmol/L)group,DMSO group.Cell counting Kit-8(CCK-8)assay was used to assess proliferation ability of HA-VSMC,and Transwell assay was used to examin the migration ability of HA-VSMC.Western blot was used to detect the expression of autophagy related proteins including p62,LC3,mammalian target of rapamycin(mTOR)and phosphorylated mTOR(p-mTOR).Results After treatment with PDGF-BB,the proliferation and migration ability of HA-VSMC was notably upregulated(P<0.01),while jaranol showed a significant and concentration-dependent inhibition effect on PDGF-BB-induced proliferation and migration of HA-VSMC(P<0.05).Western blot results showed that compared with the PDGF-BB group,the protein expression levels of LC3Ⅰand mTOR were significantly decreased in PDGF-BB+jaranol(40μmol/L)group(P<0.05);The expression levels of LC3Ⅱ/LC3Ⅰ,p62 and p-mTOR proteins were significantly increased(P<0.05),and the effect of jaranol(40μmol/L)and compound C on AMPK/mTOR signaling pathway of HA-VSMC induced by PDGF-BB was consistent.Conclusion Jaranol can inhibit the abnormal proliferation and migration of HA-VSMC invoked by PDGF-BB,its mechanism may be related to its inhibition effect of PDGF-BB-induced autophagy of HA-VSMC by regulating AMPK/mTOR signaling pathway.