黄芪多糖修复LPS诱导的内皮祖细胞功能损伤机制的实验研究

Effect of Astragalus Polysaccharides on LPS-induced EPCs Functional Damage

目的运用microRNA芯片方法检测黄芪多糖对脂多糖(LPS)刺激下人内皮祖细胞(EPCs)细胞功能损伤的诱导作用及相关的miRNA表达变化,分析其分子机制。方法将EPCs分为3组:对照组、LPS处理组(LPS组)和黄芪多糖预处理组(APS组),进行miRNA表达谱分析与验证,并利用生物信息方法分析miRNA调控的靶基因功能与信号通路。结果有15个miRNA与LPS诱导的细胞功能损伤相关。19个miRNA在黄芪多糖预处理后表达发生了改变。7个miRNA分子即hsa-miR-146a-5p、hsa-miR-145-5p、hsa-miR-7977、hsa-miR-19a-3p、hsa-miR-377-3p、hsa-miR-7114-5p及hsa-miR-424-5p在LPS组与APS组比较,差异有统计学意义(P<0.05)。其中3个miRNA经RT-PCR验证结果与芯片结果相符。7个miRNA的靶基因主要参与蛋白泛素化调节等基因本体(GO)功能分类以及PI3K-Akt等信号通路。结论 APS对LPS诱导的EPCs功能损伤有保护修复作用,该作用可能与miRNA分子的表达改变相关。

Objective To analyze the relative miRNA network of Astragalus Polysaccharides’(APS) protective effect on lipopolysaccharide(LPS)-induced functional damage of endothelial progenitor cells(EPCs). Methods For miRNA expression profile analysis, endothelial progenitor cells were divided into three groups: Control group, LPS treatment group, and APS pretreatment group. Real-time PCR were used for microarray data validation. Bioinformatics methods were used for miRNA network analysis. Results Totally 15 miRNAs were found to be associated with LPS-induced cellular functional impairment. Expressions of 19 miRNAs were changed after APS pretreatment with LPS stimulation. Seven miRNAs including hsa-miR-146 a-5 p, hsa-miR-145-5 p, hsa-miR-7977, hsa-miR-19 a-3 p, hsa-miR-377-3 p, hsa-miR-7114-5 p, and hsa-miR-424-5 p, showed significant differences between the LPS group and the APS pretreatment group(P<0.05). Expressions of 3 miRNAs verified by RT-PCR were consistent with the microarray results. GO analysis found that mRNA targets of the 7 miRNAs were enriched in protein ubiquitination functional group and PI3 K-Akt signaling pathway. Conclusions APS had protective effects on LPS-induced functional damage of EPCs. The molecular mechanism of this effect was related to expression changes of miRNAs.