摘要
目的探讨miR-133a对增生性瘢痕增殖水平及转化生长因子-β1(transforming growth factor-β,TGF-β1)、Ⅰ型胶原(collagenⅠ, COL1)、基质金属蛋白酶2(matrix metalloproteinase 2, MMP2)表达的影响。方法自2016年8月至2019年8月,选取北部战区总医院烧伤整形科的30例增生性瘢痕患者的组织为观察组;6例皮瓣移植手术中弃用的正常皮肤组织作为对照组。通过实时荧光定量PRC检测观察组和对照组中mi R-133a和TGF-β1的表达水平,分析mi R-133a与TGF-β1表达的相关性。荧光素酶报告基因实验检测mi R-133a对于TGF-β1的靶向结合作用。将观察组细胞分为3组,分别用阴性对照RNA+空载体、mi R-133a+空载体、miR-133a+TGF-β1转染,使用细胞计数试剂盒-8(CCK-8)检测增生性瘢痕成纤维细胞的增殖水平。Western blot和实时荧光定量PRC检测细胞中mi R-133a、TGF-β1、COL1和MMP2的表达。结果观察组中miR-133a的RNA表达量明显低于对照组(P<0.05)。观察组中TGF-β1的mRNA表达量明显高于对照组(P<0.05)。观察组中mi R-133a的表达与TGF-β1的表达呈负相关,下调TGF-β1的活性。miR-133a转染能够下调增生性瘢痕成纤维细胞的增殖水平,TGF-β1能够逆转mi R-133a对于增生性瘢痕成纤维细胞增殖的抑制作用,转染TGF-β1后,TGF-β1、COL1和MMP2的表达水平恢复。结论 mi R-133a能够抑制增生性瘢痕成纤维细胞的增殖,下调TGF-β1、COL1和MMP2的表达水平。
Objective To investigate the effect of miR-133 a on the proliferation of hypertrophic scar and the expression of transforming growth factor-β1(TGF-β1), collagen I(COL1) and matrix metalloproteinase 2(MMP2).Methods The hypertrophic scars from30 patients between August 2016 and 2019 were selected as observation group. The normal skin from 6 patients were selected as control group. The expression of miR-133 a and TGF-β1 in observation group and control group was detected by real-time PCR. The correlation between mi R-133 a and TGF-β1 expression was analyzed. The targeted binding of miR-133 a and TGF-β1 was detected by luciferase reporter gene assay. The cells in observation group were divided into 3 groups and transfected with negative control RNA + empty vector,mi R-133 a + empty vector and miR-133 a + TGF-β1 respectively. The proliferation of hypertrophic scar fibroblasts was detected by cell counting kit-8(CCK-8). The expression of mi R-133 a, TGF-β1, COL1 and MMP2 was detected by Western blot and real-time PCR.Results The RNA expression of miR-133 a in observation group was significantly lower than that in control group. The RNA expression of TGF-β1 in observation group was significantly higher than that in control group. The expression of mi R-133 a in observation group was negatively correlated with the expression of TGF-β1. The mi R-133 a could bind to the 3’UTR region of TGF-β1 and down regulate the activity of TGF-β1. The transfection of mi R-133 a could down regulate the proliferation of hypertrophic scar fibroblasts. The TGF-β1 could reverse the inhibition of mi R-133 a on hypertrophic scar fibroblasts proliferation. After transfected with TGF-β1, the expression of TGF-β1, COL1 and MMP2 was restored.Conclusion The mi R-133 a could inhibit the proliferation of hypertrophic scar fibroblasts and down regulate the expression of TGF-β1, COL1 and MMP2.
作者
张权
白泽明
陶凯
ZHANG Quan;BAI Ze-ming;TAO Kai(Department of Bum and Plastic Surgery,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《中国美容整形外科杂志》
CAS
2021年第11期658-661,681,共5页
Chinese Journal of Aesthetic and Plastic Surgery
