摘要
目的观察苏木乙酸乙酯提取液对内皮细胞miRNA-129-5p/Beclin1信号通路及其介导的自噬的影响。方法人脐静脉内皮细胞分为空白组、模型组、苏木组、模拟剂组及苏木+模拟剂组,除空白组外,余下各组细胞采用氧化低密度脂蛋白(ox-LDL)诱导建立内皮细胞损伤模型并给予相应药物干预。分别采用光学显微镜和透射电镜检测内皮细胞形态及自噬水平,MTT法检测细胞增殖能力,荧光素酶报告基因实验检测miRNA-129-5p和Beclin1的结合活性,RT-PCR法检测miRNA-129-5p及Beclin1 mRNA的表达,Western blotting检测Beclin1、LC3Ⅰ、LC3Ⅱ蛋白的表达。结果与模型组相比,苏木组脐静脉内皮细胞的自噬水平、Beclin1蛋白的表达和LC3Ⅱ/LC3Ⅰ的比值显著增加,miRNA-129-5p的表达水平显著降低,差异均具有统计学意义(P <0.05)。结论苏木乙酸乙酯提取液通过抑制miRNA-129-5p的表达促进Beclin1基因转录后翻译水平,进而促进细胞自噬,发挥抗动脉粥样硬化的作用。
Objective:To observe the effects of sappan acetic ether extract(SAEE)on oxidized low density lipoprotein(ox LDL)induced autophagy and miRNA-129-5 p/Beclin1 signaling pathway in endothelial cells.Methods:Human umbilical vein endothelial cells were divided into the control group,model group,SAEE group,miRNA-129-5 p mimic group and SAEE+miRNA-129-5 p mimic group.Except for the control group,the remaining groups were induced by oxidized low density lipoprotein(ox LDL)to establish endothelial cell injury model and give corresponding drug intervention.The morphology and autophagy level of endothelial cells were detected by light microscope and transmission electron microscope,the proliferation ability of endothelial cells was detected by MTT method,the binding activity of miRNA-129-5 p and Beclin1 was detected by luciferase reporter gene assay,the expression of miRNA-129-5 p and Beclin1 mRNA was detected by RT-PCR,and the protein expression of Beclin1,LC3Ⅱ/LC3Ⅰratio were detected by Western blotting.Results:Compared with the model group,the autophagy level,Beclin1 protein expression level,and LC3Ⅱ/LC3Ⅰratio in the SAEE group were significantly increased,and the expression level of miRNA-129-5 p was decreased,the differences were statistically significant(P<0.05).Conclusion:SAEE promotes the level of Beclin1 gene post transcriptional translation by inhibiting the expression of miRNA-129-5 p,and then promotes autophagy and plays an anti-atherosclerotic role.
作者
李鑫峰
袁星星
周亚滨
杨建飞
王倩
Li Xinfeng;Yuan Xingxing;Zhou Yabin;Yang Jianfei;Wang qian(Guizhou University of Traditional Chinese medicine,Guizhou,Guiyang 550025,China)
出处
《中国中医急症》
2021年第4期617-621,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
黑龙江中医药管理局中医药科研项目(ZHY18-029)
黑龙江省教育厅青年创新人才培养项目(UNPYSCT-2018228)
贵州中医药大学博士启动基金(2019-98)。
