基于microRNA探讨半夏白术天麻汤内皮保护机制  

Exploration of Banxia Baizhu Tianma Decoction on endothelial protective effects based on microRNA

作　　者：姜月华[1,2] 张鹏[1] 李兆钰 姜凌宇 杨传华[2] JIANG Yue-hua;ZHANG Peng;LI Zhao-yu;JIANG Ling-yu;YANG Chuan-hua(Shandong University of Traditional Chinese Medicine,Jinan 250355.China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学,济南250355 [2]山东中医药大学附属医院,济南250014

出　　处：《中华中医药杂志》2021年第4期1995-1999,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81673807);泰山学者岗位建设资金(No.2018-35)。

摘　　要：目的:通过小RNA测序及siRNA基因敲减/过表达技术,探索半夏白术天麻汤内皮保护的作用靶点。方法:体外培养人主动脉内皮细胞(HAEC),50μg/mL氧化型低密度脂蛋白(ox-LDL)诱导细胞损伤,1h后加入2mg/mL半夏白术天麻汤继续培养24h,未处理的HAEC细胞作为正常对照组。小RNA测序筛选差异microRNA(miRNA),并基于siRNA敲减/过表达miRNA-217-5p(即miR-217),同步给予半夏白术天麻汤干预,观察miRNA-217的表达及其对细胞增殖的影响。结果:经小RNA测序,ox-LDL诱导后90个miRNA差异表达,半夏白术天麻汤干预后,共筛选出157个差异miRNA(93个上调,64个下调)(FC>2.0,P<0.05),其中60个共有差异miRNA,提示半夏白术天麻汤逆转了ox-LDL诱导的66.67%的miRNA改变,共有基因中差异最显著的是miRNA-217。miRNA-217敲减/过表达后,细胞增殖水平发生相应的变化;半夏白术天麻汤的干预效果在miRNA-217敲减后下降,而miRNA-217过表达可增强半夏白术天麻汤的药效。结论:miRNA-217是半夏白术天麻汤的重要靶点,半夏白术天麻汤通过上调miRNA-217,促进内皮细胞增殖,发挥内皮保护作用。Objective:To explore the target genes of Banxia Baizhu Tianma Decoction(BBTD)on endothelial protection based on small RNA sequencing and knockdown/overexpression by siRNA.Methods:Human aortic endothelial cells(HAECs)were cultured in vitro.The endothelial injury was induced by 50μg/mL oxidized low density lipoprotein(ox-LDL),followed by 2 mg/mL BBTD for 24 h,with untreated HAECs as normal control.Differentially expressed(DE)microRNAs(miRNAs)were screened by small RNA sequencing.Based on knock down/over expression of miRNA-217 by siRNA,BBTD was added simultaneously to assess the expression of miRNA-217 and its target genes and its effect on cell proliferation.Results:According to the data of small RNA sequencing,ox-LDL induced 90 miRNAs differentially expressed(DE);while after BBTD treatment,157 DE miRNAs(93 upregulated and 64 downregulated)(FC>2.0,P<0.05)were screened out,among which 60 were overlapped,suggesting that BBTD reversed 66.67%of miRNA changes induced by ox-LDL and the most significant difference was miRNA-217.The knockdown/overexpression of miRNA-217 leaded to the corresponding change of its target genes and cellular proliferation.The efficacy of BBTD was decreased after miRNA-217 knockdown,while miRNA-217 overexpression enhanced the effect of BBTD.Conclusion:miRNA-217 is one of the important targets of Banxia Baizhu Tianma Decoction.Banxia Baizhu Tianma Decoction demonstrates endothelial protective effects by upregulating miRNA-217 and its target genes.

关 键 词：半夏白术天麻汤 氧化型低密度脂蛋白 人主动脉内皮细胞 MICRORNA microRNA-217-5p 

分 类 号：R285[医药卫生—中药学]