冠心康调控巨噬细胞miRNA-155与SOCS1-STAT3-PDCD4生物轴抗动脉粥样硬化研究

Research of Guanxinkang regulates macrophage miRNA-155 and SOCS1-STAT3-PDCD4 biological axis against atherosclerosis

目的:探讨冠心康调控巨噬细胞信号及抗动脉粥样硬化的作用机制。方法:RAW264.7细胞按随机数字表法分为对照组(空白血清)、微小RNA-155(miR-155)mimic组、miR-155 inhibitor组、含药血清高剂量组、含药血清低剂量组5组,分别给予药物和含药血清干预后,采用RT-PCR法和Western-blot法检测各组细胞中miR-155、细胞因子信号传导抑制剂l(SOCS1)、磷酸化转录激活子3(p-STAT3)、程序性细胞凋亡因子4(PDCD4)mRNA和蛋白表达,Elisa法检测细胞分泌肿瘤坏死因子-a(TNF-α)、白细胞介素6(IL-6)、干扰素-γ(IFN-γ)等炎症因子水平。结果:与对照组比较,miR-155mimic组和含药血清组细胞中miR-155 mRNA、SOCSl mRNA和蛋白表达明显较高,miR-155 inhibitor组细胞中miR-155 mRNA、SOCSl mRNA和蛋白表达明显较低。与对照组比较,miR-155mimic组和含药血清组细胞中p-STAT3和PDCD4 mRNA和蛋白表达明显较低,miR-155inhibitor组细胞中p-STAT3、PDCD4 mRNA和蛋白表达明显较高。与对照组比较,miR-155mimic组和含药血清组细胞液中TNF-α、IL-6、IFN-γ水平明显较低,miR-155inhibitor组细胞液中炎症因子水平明显较高。结论:冠心康抗动脉粥样硬化的分子机制可能与调节miR-155进而调控SOCSl/p-STAT3/PDCD4信号通路有关。

Objective:To investigate the mechanism of Guanxinkang(GXK)regulating macrophage signal pathway and anti-atherosclerosis effects in rats.Methods:RAW264.7 cells were randomly divided into 5 groups:control group(blank serum),microRNA-155(miR-155)mimic group,miR-155 inhibitor group,drug-containing serum high-dose group,and drug-containing serum low-dose group.After intervention with drugs and drug-containing serum,the expression of miR-155,SOCS1 and p-STAT3,PDCD4 were detected by Real-time PCR and Western-blot.Elisa assay was used to detect levels of inflammatory factors TNF-α,IL-6,IFN-γin Cell fluid.Results:Compared with the normal group,the expression of miR-155 and SOCS1 in miR-155mimic group and drug-containing serum group were significantly higher,the expression of miR-155 and SOCS1 in miR-155inhibitor group were significantly lower.Compared with the normal group,the expression of p-STAT3 and PDCD4 in miR-155mimic group and drug-containing serum group were significantly lower,the expression of p-STAT3 and PDCD4 in the miR-155inhibitor group were significantly higher.Compared with the normal group,the levels of TNF-α,IL-6 and IFN-γin the cell fluid of miR-155mimic group and drug-containing serum group were significantly lower,and those in the miR-155 inhibitor group was significantly higher.Conclusion:The mechanism of Guanxinkang against atherosclerosis may be related to regulating SOCS1/p-STAT3/PDCD4 signaling pathway through the regulation of miR-155.