文章摘要
熊 亮,胡 寅,王立夫,曹咬脐,钱莉丽,刘泾科.miRNA⁃199a⁃5p在热损伤人皮肤成纤维细胞中的表达及其功能研究[J].南京医科大学学报,2021,(8):1160~1165
miRNA⁃199a⁃5p在热损伤人皮肤成纤维细胞中的表达及其功能研究
Expression and function of miRNA⁃199a⁃5p in human skin fibroblasts after thermal injury
投稿时间:2020-12-23  
DOI:doi:10.7655/NYDXBNS20210808
中文关键词: 皮肤成纤维细胞  热损伤  miRNA⁃199a⁃5p  qRT⁃PCR  细胞增殖  细胞迁移与侵袭
英文关键词: human skin fibroblasts  thermal injury  miRNA⁃199a⁃5p  qRT⁃PCR  cell proliferation  cell migration and invasion
基金项目:苏州市吴江区科技局科研项目(WWK201615)
作者单位
熊 亮 南京医科大学附属江苏盛泽医院整形烧伤科江苏 苏州 215228 
胡 寅 南京医科大学第一附属医院整形烧伤科江苏 南京 210029 
王立夫 南京医科大学第一附属医院整形烧伤科江苏 南京 210029 
曹咬脐 南京医科大学附属江苏盛泽医院整形烧伤科江苏 苏州 215228 
钱莉丽 南京医科大学附属江苏盛泽医院整形烧伤科江苏 苏州 215228 
刘泾科 南京医科大学附属江苏盛泽医院整形烧伤科江苏 苏州 215228 
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中文摘要:
      目的:观察热损伤人皮肤成纤维细胞中miRNA?199a?5p表达量的变化,以及对皮肤成纤维细胞增殖、迁移、侵袭功能的影响。方法:分别用人皮肤成纤维细胞株(HSF)隔水52 ℃水浴30 s、60 s制造细胞热损伤模型,对比观察细胞热损伤状态;用四甲基偶氮唑盐(MTT)检测热损伤24 h、48 h后细胞的增殖率;用qRT?PCR检测热损伤24 h、48 h后细胞miRNA?199a?5p表达量的变化情况;用划痕和Transwell测试细胞的迁移、侵袭功能;用模拟物提高热损伤细胞中miRNA?199a?5p的表达量,检测热损伤细胞的增殖和侵袭情况。结果:52 ℃ 30 s可以很好地制造人皮肤成纤维热损伤模型;与对照组比较,热损伤后miRNA?199a?5p表达量下调[24 h,(2.67±0.35)%,P<0.001;48 h,(18.42±0.44)%,P<0.001],细胞增殖显著受到抑制[24 h,(17.10±3.21)%,P<0.002;48 h,(25.93±1.74)%,P<0.001],细胞的迁移功能显著下降[(35.35±7.07)%,P<0.001],侵袭功能亦被显著抑制[(57.14±9.52)%,P<0.007]。通过模拟物提高热损伤细胞中miRNA?199a?5p的表达量后,细胞的增殖显著上调[(136.55±6.14)%,P<0.003],细胞的侵袭能力增强[(196.88±15.63)%,P<0.003]。结论:热损伤人皮肤成纤维细胞中miRNA?199a?5p的表达量与细胞增殖、迁移和侵袭正相关,提高热损伤细胞中miRNA?199a?5p的表达量可以帮助热损伤细胞快速转归。
英文摘要:
      Objective:This study aims to observe the changes of miRNA?199a?5p expression in human skin fibroblasts with thermal injury and its effects on the proliferation,migration and invasion of skin fibroblasts. Methods:Cell models of thermal injury were established using human skin fibroblast cell line by bathing in water at 52℃ for 30 s and 60 s,respectively,and the thermal injury of cells was observed and compared. The proliferation rate of cells 24 h and 48 h after thermal injury was detected by MTT assay. The changes of miRNA?199a?5p expression in the cells 24 and 48 h after thermal injury were determined through qRT?PCR. Cell migration and invasion were detected using wound?healing assay and Transwell assay. The expression of miRNA?199a?5p in thermally injured cells was increased using mimics,and the proliferation and invasion of thermally injured cells were detected. Results:Bathing in water at 52 ℃ for 30 s could successfully establish a thermal injury model in human skin fibroblasts. After thermal injury,miRNA?199a?5p expression was down?regulated[24 h,(2.67±0.35)%,P<0.001;48 h,(18.42±0.44)%,P<0.001],cell proliferation was significantly inhibited[24 h,(17.10±3.21)%,P<0.002;48 h,(25.93±1.74)%,P<0.001],cell migration decreased significantly[(35.35±7.07)%,P<0.001],and cell invasion was also significantly inhibited[(57.14±9.52)%,P<0.007]. After enhancing miRNA?199a?5p expression in the thermally injured cells using mimics,the proliferation of the cells increased significantly[(136.55±6.14)%,P<0.003],and the invasion ability of the cells was enhanced[(136.55±6.14)%,P<0.003]. Conclusion:The expression level of miRNA?199a?5p in human skin fibroblasts with thermal injury is positively correlated with cell proliferation,migration and invasion. Increasing the expression level of miRNA?199a?5p in thermally injured cells can help it rapid return.
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