低幅度机械牵张力诱导肌成纤维细胞再分化的作用  

EFFECTOF LOW-AMPLITUDE MECHANICALSTRAINININDUCING THE REDIFFERENTIATION OF MYOFIBROBLASTS

作　　者：安玉[1] 郝荣安 匡瑞霞[1] 胡春楠[1] 陈璐[1] 王志国[1] AN Yu;HAO Rongan;KUANG Ruixia;HU Chunnan;CHEN Lu;WANG Zhiguo(Department of Burns and Plastic Surgery,The Affiliated Hospital of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属医院烧伤整形外科,山东青岛266071 [2]青岛元美整形医院

出　　处：《青岛大学学报（医学版）》2021年第3期432-436,共5页Journal of Qingdao University(Medical Sciences)

基　　金：山东省自然科学基金资助项目(ZR2017MH083)。

摘　　要：目的探讨低幅度机械牵张力对肌成纤维细胞再分化的诱导作用及增生性瘢痕的转归机制。方法体外培养健康人皮肤成纤维细胞,应用10%幅度牵张力诱导构建肌成纤维细胞模型。实验分为阴性对照组、阳性对照组、实验组,以人正常成纤维细胞为阴性对照组,应用2%幅度牵张力体外培养;以肌成纤维细胞作为实验组和阳性对照组,分别应用2%、10%幅度牵张力继续体外培养,诱导细胞再分化。实验第7天,应用CCK-8法检测细胞增殖能力,SYBR Green qPCR法检测整合素β1、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原的mRNA表达,ELISA法检测TGF-β1、Ⅰ型胶原含量;Western-Blot法检测整合素β1、α-SMA蛋白表达量。结果与阴性对照组比较,实验组肌成纤维细胞的增殖能力增高(F=67.75,P<0.05),细胞内TGF-β1、Ⅰ型胶原的蛋白含量、α-SMA的蛋白表达及各指标的mRNA表达水平均增高(F=132.64~981.60,P<0.05),细胞内整合素β1 mRNA和蛋白表达水平差异无统计学意义(P>0.05);阳性对照组和实验组肌成纤维细胞内α-SMA mRNA和蛋白表达差异无统计学意义(P>0.05);而阳性对照组肌成纤维细胞内TGF-β1、Ⅰ型胶原的mRNA和蛋白含量与整合素β1的mRNA和蛋白表达水平均较实验组增高,差异有统计学意义(P<0.05)。结论低幅度牵张力不能诱导肌成纤维细胞再分化,可能存在其他机制介导了张力诱导肌成纤维细胞再分化的过程。Objective To explore the effect of low-amplitude mechanical strain in inducing the redifferentiation of myofibroblasts,and to investigate the prognostic mechanism of hyperplastic scars.Methods Fibroblasts from the skin of healthy people were cultured in vitro,and a model of myofibroblasts was established by applying a strain at an amplitude of 10%.The experiment included three groups,i.e.,negative control group,positive control group,and experimental group.Normal human fibroblasts were taken as the negative control group,which were subjected to a strain at an amplitude of 2%when cultured in vitro.Myofibroblasts,which were subjected to a strain at amplitudes of 2%and 10%to induce cell redifferentiation during in vitro culture,were taken as the experimental group and positive control group,respectively.On day 7 of the experiment,the CCK-8 was used to determine the proliferative capacity of cells;SYBR Green qPCR assay was used to determine the mRNA expression of integrinβ1,transforming growth factorβ1(TGF-β1),alpha-smooth muscle actin(α-SMA),and typeⅠcollagen;enzyme-linked immunosorbent assay was used to determine the content of TGF-β1 and typeⅠcollagen;Western Blot was used to determine the protein expression of integrinβ1 andα-SMA.Results Compared with the negative control group,the experimental group had significantly increased proliferative capability of myofibroblasts(F=67.75,P<0.05)and significantly increased protein content of TGF-β1 and typeⅠcollagen,protein expression ofα-SMA,and mRNA expression of the three indicators in the cells(F=132.64-981.60,P<0.05),but there were no significant differences in the mRNA and protein expression levels of integrinβ1 in cells between the two groups(P>0.05).There were no significant differences in the mRNA and protein expression ofα-SMA in myofibroblasts between the positive control group and the experimental group(P>0.05),while the former group had significantly higher mRNA expression and protein content of TGF-β1 and typeⅠcollagen and mRNA and protein

关 键 词：机械张力 肌成纤维细胞 成纤维细胞 细胞分化 瘢痕 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]