miR-205-5p/PRKCA/p38信号通路在淫羊藿苷抑制动脉粥样硬化中的介导作用  

Mediating effect of MiR-205-5p/PRKCA/p38 signal path on atherosclerotic inhibitory

作　　者：徐瑞 黄鹏 张祎冰 任立群[2] XU Rui;HUANG Peng;ZHANG Yi-Bing(Norman Bethune College of Medicine,Changchun 130021,Jilin,China)

机构地区：[1]吉林大学白求恩医学部,吉林长春130021 [2]吉林大学药学院实验药理与毒理学教研室,吉林长春130021 [3]吉林大学第一医院眼科,吉林长春130021

出　　处：《中国老年学杂志》2020年第10期2169-2176,共8页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(No.81773934)。

摘　　要：目的探讨miR-205-5p/蛋白激酶Cα(PRKCA)/p38信号通路在淫羊藿苷(ICA)抑制动脉粥样硬化(AS)中的介导作用。方法首先通过整体动物实验研究ICA对载脂蛋白(Apo)E^-/-小鼠的抗AS作用;然后取小鼠胸主动脉组织进行基因芯片检测,再通过生物信息分析找到有意义的miRNA及其作用的靶基因,级联通路;最后应用定量聚合酶链反应(qPCR)及Western印迹方法对所构建的通路在体内外实验标本中进行验证。结果在整体动物实验中,血脂水平和HE染色结果表明,ICA具有抗AS作用;通过对基因芯片检测结果分析找到目标miR-205-5p,通过生物信息分析预测其靶基因为PRKCA,并级联p38信号通路分子,构建miR-205-5p/PRKCA/p38信号通路;qPCR及Western印迹结果显示,在ApoE^-/-小鼠胸主动脉及氧化低密度脂蛋白(ox-LDL)刺激血管平滑肌细胞(VSMCs)中,模型组miR-205-5p表达显著下降(P<0.001),PRKCA mRNA及蛋白表达显著升高,p38蛋白磷酸化显著增加(P<0.001);ICA干预后,miR-205-5p表达显著升高(P<0.001),PRKCA mRNA及蛋白表达显著下降(P<0.001),p38蛋白磷酸化显著减少(P<0.001),且具有量效关系;ICA对ox-LDL刺激VSMCs的增殖和迁移具有显著抑制作用(P<0.001或P<0.05)。结论ICA具有抗AS作用,其机制可能是通过提高miR-205-5p表达,下调PRKCA mRNA及蛋白表达,进而减少p38蛋白磷酸化,并抑制VSMCs增殖与迁移,从而发挥其抗AS作用。Objective To explore the effects of miR-205-5p/PRKCA/p38 signaling pathway in the inhibition of icariin on atherosclerosis by whole animal experiments and in vitro cell experiments.Methods First,the whole animal experiment was used to study the anti-atherosclerotic effect of ICA on ApoE^-/- mice;Then meaningful miRNAs and their target genes,cascade pathways were found through the analysis of biological information;Finally,qPCR and Western blot were used to verify the constructed pathways in vivo and in vitro experimental specimens.Results In whole animal experiments,blood lipid levels and HE staining results indicated that ICA had an anti-atherosclerotic effect;The target miR-205-5p was found by analyzing the results of the gene chip analysis.The target gene was predicted to be PRKCA mRNA by biomarker analysis,and the p38 signal pathway signal molecules were cascaded to construct the miR-205-5p/PRKCA/p38 signal pathway;qPCR and Western blot results showed that in the thoracic aorta tissue of ApoE^-/- mice and VSMCs stimulated by ox-LDL,miR-205-5p expression in the model group was significantly reduced(P<0.001),PRKCA mRNA and protein significantly increased expression and significantly increased p38 phosphorylation(P<0.001);After ICA intervention,miR-205-5p expression was significantly increased(P<0.001),PRKCA mRNA and protein expression were significantly decreased(P<0.001),p38 phosphorylation was significantly reduced(P<0.001),and there was a dose-effect relationship;ICA significantly inhibited the proliferation and migration of VSMCs stimulated by ox-LDL in mice(P<0.001 or P<0.05).Conclusions ICA has anti-atherosclerotic effect.The mechanism might be through increasing miR-205-5p expression,down-regulating PRKCA mRNA and protein expression,thereby reducing p38 phosphorylation and inhibiting VSMCs proliferation and migration,thereby exerting its anti-atherosclerotic effect.

关 键 词：动脉粥样硬化 淫羊藿苷 miR-205-5p 蛋白激酶Cα(PRKCA) P38 

分 类 号：R541.4[医药卫生—心血管疾病]