人脐带间充质干细胞对增生性瘢痕成纤维细胞的抑制作用及机制研究  

Inhibitory effect and mechanism of human umbilical cord mesenchymal stem cells on hypertrophic scar fibroblasts

作　　者：王琨皓 舒俊森 郭伶俐[2] WANG Kunhao;SHU Junsen;GUO Lingi(Chinese PLA Medical School,Beijing 100853,China;Department of Plastic and Reconstructive Surgery,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院,北京100853 [2]解放军总医院第一医学中心整形修复外科,北京100853

出　　处：《解放军医学院学报》2024年第11期1184-1190,共7页Academic Journal of Chinese PLA Medical School

基　　金：北京市自然科学基金项目(7222170);省部级课题。

摘　　要：背景增生性瘢痕是病理性瘢痕的一种,常影响患者的身心生活质量。间充质干细胞在抗瘢痕治疗中具有巨大的潜力。目的研究人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)对体外培养的人增生性瘢痕成纤维细胞(hypertrophic scar fibroblasts,HSFBs)的影响作用及可能机制。方法体外培养的hUC-MSCs与人HSFBs接种细胞数量比例分别为0.5∶1、1∶1、2∶1(0.5∶1组、1∶1组、2∶1组),另外设置只接种HSFBs的对照组、转化生长因子β1(transforming growth factorβ1,TGF-β1)干预组(TGF-β1浓度为5 ng/mL)以及TGF-β1和hUC-MSCs共同干预组(hUC-MSCs与HSFBs数量比为1∶1)。利用Transwell小室共培养,一定时间后取各组下室中HSFBs及其上清液,CCK-8法测定HSFBs增殖率;羟脯氨酸试剂盒检测各组HSFBs上清液中羟脯氨酸含量;划痕实验检测HSFBs迁移率;RT-qPCR检测HSFBs中TGF-β1、Smad3、COL-Ⅰ、COL-Ⅲ、PI3K、AKT的转录水平。结果与对照组相比,0.5∶1组、1∶1组、2∶1组均能够抑制人HSFBs增殖(P<0.05),其中1∶1组抑制增殖的作用最显著且能够抑制HSFBs胶原分泌(P<0.05);划痕12 h后0.5∶1组、1∶1组、2∶1组与对照组比较,细胞迁移的差异无统计学意义(P>0.05),24 h后1∶1组、2∶1组细胞迁移能力弱于对照组和0.5∶1组(P<0.05);0.5∶1组、1∶1组、2∶1组中TGF-β1、Smad3、COL-Ⅰ、COL-Ⅲ的转录水平相比对照组降低(P<0.05);1∶1组的PI3K转录水平低于0.5∶1组(P<0.05);AKT的转录水平在对照组、0.5∶1组、1∶1组、2∶1组间差异均无统计学意义(P>0.05)。TGF-β1干预组相比于对照组,能够促进增生性瘢痕成纤维细胞增殖(P<0.01)、促进胶原分泌(P<0.01)、促进细胞迁移(P<0.05),TGF-β1干预组的TGF-β1、Smad3、COL-Ⅰ、COL-Ⅲ、PI3K、AKT转录水平相比对照组升高(P<0.01),hUC-MSCs干预能够抑制TGF-β1促进HSFBs增殖、迁移、胶原分泌、相关基因转录的作用(P<0.01)。结论hUC-MSCs可Background Hypertrophic scar(HS)is a type of pathological scar that often affects the physical and mental quality of life of patients.In recent years,the research of mesenchymal stem cells has become more and more in-depth,and they have great potential in anti-scar therapy.Objective To study the effect and possible mechanism of human umbilical cord mesenchymal stem cells(hUC-MSCs)on human hypertrophic scar fibroblasts(HSFBs)cultured in vitro.Methods The hUC-MSCs cultured in vitro and human HSFBs were divided into three groups according to the proportion of inoculated cells:0.5:1,1:1 and 2:1.In addition,the control group inoculated with HSFBs only,the transforming growth factor beta 1(TGF-β1)intervention HSFBs group(TGF-β1 concentration was 5 ng/mL)and the TGF-β1 and hUC-MSCs co-intervention group(ratio of hUC-MSCs to HSFBs was 1:1)were set up.After a certain period of time,hypertrophic scar fibroblasts and their supernatant were taken from six groups,and the proliferation rate of fibroblasts was measured by CCK-8.The collagen content in supernatant of hypertrophic scar fibroblasts in each group was detected by hydroxyproline method.Fibroblast mobility was detected by scratch test.The transcription levels of TGF-β1,Smad3,COL-Ⅰ,COL-Ⅲ,PI3K and AKT in fibroblasts were detected by RT-qPCR.Results Compared with the control group,the 0.5:1,1:1,2:1 groups could inhibit the proliferation of human hypertrophic scar fibroblasts(P<0.05),and the 1:1 group had the most obvious inhibitory effect on proliferation and could inhibit the collagen secretion of fibroblasts(P<0.05).There was no significant difference in cell migration between the 0.5:1,1:1,2:1 groups and the control group after 12 h(P>0.05),and the cell migration ability of the 1:1,2:1 groups was weaker than that of the control group and the 0.5:1 group after 24 h(P<0.05).The transcription levels of TGF-β1,Smad3,COL-Ⅰand COL-Ⅲin 0.5:1,1:1 and 2:1 groups were lower than those in control group(P<0.05).The PI3K transcription level in 0.5:1 group was lower th

关 键 词：脐带间充质干细胞 增生性瘢痕 成纤维细胞 转化生长因子Β1 纤维化 

分 类 号：R622.9[医药卫生—整形外科]