Roc-A通过调节自噬和活性氧积累抑制血管平滑肌细胞增殖迁移

Roc-A Inhibits Proliferation and Migration of Vascular Smooth Muscle Cells by Regulating Autophagy and Reactive Oxygen Species Accumulation

该研究探讨了在PDGF-BB诱导下, Roc-A对血管平滑肌细胞(VSMCs)增殖和迁移的抑制作用及调控机制。用25 ng/mL PDGF-BB刺激大鼠A7r5细胞建立细胞损伤模型,再将细胞分为对照组(NC)、PDGF-BB模型组、PDGF-BB+10 nmol/L Roc-A组、PDGF-BB+25 nmol/L Roc-A组和PDGF-BB+50 nmol/L Roc-A组。通过CCK8和Transwell分别检测细胞活力和增殖迁移能力;流式细胞术检测细胞内EdU掺入情况和细胞周期水平;DCFH-DA荧光探针检测活性氧积累水平;Western blot检测PCNA、LC3B、P62蛋白表达水平。结果显示, PDGF-BB处理使细胞活力显著增加,并促进了细胞增殖和迁移。Roc-A处理下调了PDGF-BB诱导的细胞活性和PCNA表达,抑制了细胞增殖和迁移;Roc-A使细胞周期受阻, S期细胞比例呈剂量依赖性的减少。同时, Roc-A逆转了PDGF-BB诱导的活性氧积累和LC3B II、P62自噬相关蛋白表达。5 mmol/L 3-MA与25 nmol/L Roc-A处理均抑制了平滑肌细胞增殖迁移。这说明Roc-A抑制PDGF-BB诱导的大鼠VSMCs增殖和迁移,其可能是通过下调PDGF-BB诱导的自噬和氧化应激发挥作用的,提示了其对内膜增生引起的再狭窄具有治疗作用。

This study investigated the inhibitory effect of Roc-A on the proliferation and migration of VSMCs(vascular smooth muscle cells)induced by PDGF-BB and the regulatory mechanism.The rat A7r5 cells were stimulated with 25 ng/mL PDGF-BB to establish a cell injury model.The cells were divided into five groups:control group(NC),PDGF-BB model group,PDGF-BB+10 nmol/L Roc-A group,PDGF-BB+25 nmol/L Roc-A group and PDGF-BB+50 nmol/L Roc-A group.The cell viability and proliferation migration ability seperately were measured by CCK8 and Transwell;intracellular EdU admixture and cell cycle level were measured by flow cytometry;reactive oxygen species accumulation was measured by DCFH-DA fluorescent probe;PCNA,LC3B and P62 protein expression levels were measured by Western blot.The results showed that PDGF-BB treatment led to a significant increase in cell viability and promoted cell proliferation and migration.Roc-A treatment down-regulated PDGF-BB-induced cell activity and PCNA expression,and inhibited cell proliferation and migration.The cell cycle was blocked by Roc-A and the proportion of S-phase cells was reduced in a dose-dependent manner.Meanwhile,Roc-A reversed PDGF-BB-induced reactive oxygen species accumulation and the expression of LC3B II and P62 autophagy-related proteins.Both 5 mmol/L 3-MA and 25 nmol/L Roc-A treatments inhibited smooth muscle cell proliferation and migration.This suggests that Roc-A inhibits PDGF-BB-induced proliferation and migration of VSMCs in rats.This inhibitory effect may be mediated by down-regulation of PDGF-BB-induced autophagy and oxidative stress.It implies a therapeutic effect of Roc-A on endothelial proliferation-induced restenosis.