过表达miR-21-5p脂肪干细胞外泌体下调JAK/STAT通路调控增生性瘢痕形成的机制探讨  

Mechanism of overexpression of adipose derived stem cells exosomal miR-21-5p inhibiting JAK/STAT pathway in regulation of hypertrophic scar formation

作　　者：娄艳红 王军杰 葛玉颖 许天人 吴利永 Lou Yanhong;Wang Junjie;Ge Yuying;Xu Tianren;Wu Liyong(Department of Plastic Surgery,Henan Provincial Third People’s Hospital,Zhengzhou 450000,China)

机构地区：[1]河南省直第三人民医院整形外科,郑州450000

出　　处：《中国实用医刊》2025年第1期21-26,共6页Chinese Journal of Practical Medicine

基　　金：河南省医学科技攻关计划项目(LHGJ20230665)。

摘　　要：目的探讨过表达微小RNA-21-5p(miR-21-5p)脂肪干细胞外泌体下调Janus激酶/信号传导及转录激活蛋白(JAK/STAT)通路调控增生性瘢痕形成的机制。方法实验性研究。实验时间为2023年3月至8月。选取中国科学院细胞库典型培养物保藏中心提供的人增生性瘢痕成纤维细胞(hHSFs),设立hHSFs组、hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组,培养72 h。培养结束后,应用CCK-8试剂盒检测细胞活力,甲基紫染色检测细胞单克隆形成数目,FAScan流式细胞仪评估细胞凋亡状态,逆转录聚合酶链式反应法及蛋白印迹法测定细胞Janus激酶(JAK)、信号传导及转录激活蛋白(STAT)信使RNA(mRNA)和蛋白表达水平。结果hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组光密度(OD)值、存活率低于hHSFs组,hHSFs组+外泌体(miR-21-5p)组OD值、存活率低于hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组(P<0.05)。hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组单克隆形成数目低于hHSFs组,hHSFs组+外泌体(miR-21-5p)组单克隆形成数目低于hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组(P<0.05)。hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组凋亡率高于hHSFs组,hHSFs组+外泌体(miR-21-5p)组凋亡率高于hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组(P<0.05);hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组COLⅠ、COLⅢ蛋白水平低于hHSFs组,hHSFs组+外泌体(miR-21-5p)组单COLⅠ、COLⅢ蛋白水平低于hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组(P<0.05)。hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组、hHSFs组+外泌体(miR-21-5p)组JAK、STAT mRNA和蛋白表达低于hHSFs组,hHSFs组+外泌体(miR-21-5p)组单JAK、STAT mRNA和蛋白表达水平低于hHSFs组+外泌体组、hHSFs组+外泌体(miR-NC)组(P<0.05)。结论miR-21-5pObjectiveTo investigate the mechanism of regulation of hypertrophic scar formation by overexpression of adipose derived stem cells exosomal microRNA-21-5p(miR-21-5p)inhibiting Janus kinase-signal transducer and activator of transcription(JAK/STAT)pathway.MethodsFrom March to August 2023,a experimental study was conducted on 4 groups including human hypertrophic scar fibroblasts(hHSFs)group,hHSFs+exosome group,hHSFs+exosomal miR-21-5p negative control(miR-NC)group,referred to as hHSFs+exosome(miR-NC)group,and hHSFs+exosome(miR-21-5p)group;the hHSFs wer provided by China Center for Type Culture Colleccion,and the cells were cultured for 72 hours.After the culture,cell viability was assessed using a cell counting kit-8;the number of cell colonies was determined through methyl violet staining;apoptosis was evaluated using a FAScan flow cytometry;and the expressions of Janus kinase(JKA)messenger RNA(mRNA)and signal transducer and activator of transcription(STAT)mRNA were determined by reverse transcription polymerase chain reaction;while expressions of JAK protein and STAT protein were determined using western blotting.ResultsThe optical density(OD)and survival rate of the hHSFs+exosome group,hHSFs+exosome(miR-NC)group and hHSFs+exosome(miR-21-5p)group were lower than those of the hHSFs group(P<0.05).The OD and survival rate of the hHSFs+exosome(miR-21-5p)group were lower than those of the hHSFs+exosome group and hHSFs+exosome(miR-NC)group(P<0.05).The numbers of monoclonal colonies in the hHSFs+exosome group,hHSFs+exosome(miR-NC)group,and hHSFs+exosome(miR-21-5p)group were lower than that in the hHSFs group(P<0.05);and the number of monoclonal colonies in the hHSFs+exosome(miR-21-5p)group was lower than those in the hHSFs+exosome group and hHSFs+exosome(miR-NC)group(P<0.05).The apoptosis rates in the hHSFs+exosome group,hHSFs+exosome(miR-NC)group,and hHSFs+exosome(miR-21-5p)group were higher than that in the hHSFs group(P<0.05).The apoptosis rate in the hHSFs+exosome(miR-21-5p)group was higher than that in the hHSFs+e

关 键 词：脂肪干细胞 外泌体 微小RNA-21-5p Janus激酶/信号传导及转录激活蛋白通路 增生性瘢痕 

分 类 号：R622[医药卫生—整形外科]