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目的:分离提取甘草水煎剂中的微小核糖核酸(miRNA),并初步探索其对人外周血单个核细胞(PBMC)的影响。方法:将甘草干燥药材按照常规方法制备水煎剂,浓缩干燥后采用通用植物microRNA快速提取试剂盒提取其中的miRNA,所得miRNA经DNaseⅠ处理后进行电泳鉴定;分别利用甘草水提物、甘草酸、甘草次酸和甘草miRNA处理体外培养的健康人PBMC,24 h后,对各组进行细胞计数,并观察细胞形态学的变化;分别利用antiCD3、anti-CD56、anti-HLA-DR单克隆抗体染色,检测PBMC中不同细胞亚群的变化情况。结果:电泳结果表明,从甘草干燥药材的水煎剂中确实能够提取到miRNA,进一步印证了miRNA的稳定性。对体外培养PBMC作用结果表明,与空白对照组比较,经甘草miRNA刺激的PBMC发生了明显的抱团,细胞数量增多,HLA-DR+细胞的比例显著增加。结论:本实验成功地从甘草干燥药材的水煎剂中提取到了甘草miRNA,甘草miRNA对PBMC的生长具有促进作用,并能显著增加HLA-DR+细胞亚群的比例。
Objective: To isolate and extract miRNA from licorice root decoction and to explore its effect on human peripheral blood mononuclear cells (PBMCs). Methods: The decoction of licorice dry herbs was prepared according to the conventional method. The miRNAs were extracted by universal plant microRNA rapid extraction kit after being concentrated and dried. The resulting miRNAs were identified by electrophoresis after treatment with DNaseⅠ. Glycyrrhetinic acid and licorice miRNA were used to treat PBMCs cultured in vitro. After 24 h, the cells were counted and the morphological changes were observed. Anti-CD3, anti-CD56 and anti-HLA-DR monoclonal antibody , Detect the changes of different cell subgroups in PBMC. Results: The electrophoresis results showed that miRNA could indeed be extracted from the decoction of licorice dry herbs, further confirming the stability of miRNA. The results of in vitro culture of PBMC showed that compared with the blank control group, licorice-miRNA-stimulated PBMCs developed obvious tumbling, the number of cells increased, and the proportion of HLA-DR + cells increased significantly. Conclusion: In this study, licorice miRNAs were successfully extracted from the decoct of licorice dry herbs, and licorice miRNAs can promote the growth of PBMCs and significantly increase the proportion of HLA-DR + cell subsets.