Page 58 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第8期
·1160 · Journal of Nanjing Medical University（Natural Sciences） 2021年8月

·基础医学·

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

熊 亮 ，胡 寅 ，王立夫 ，曹咬脐 ，钱莉丽 ，刘泾科 1
2*
1
1
2
1
南京医科大学附属江苏盛泽医院整形烧伤科，江苏 苏州 215228；南京医科大学第一附属医院整形烧伤科，江苏 南京
1 2
210029
［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miR⁃
NA⁃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。
［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
［中图分类号］ R329.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）08⁃1160⁃06
doi：10.7655/NYDXBNS20210808

Expression and function of miRNA⁃199a⁃5p in human skin fibroblasts after thermal injury

2*
1
2
1
1
XIONG Liang ，HU Yin ，WANG Lifu ，CAO Yaoqi ，QIAN Lili ，LIU Jinke 1
1 Department of Plastic and Burns，Jiangsu Shengze Hospital Affiliated to Nanjing Medical University，Suzhou
2
215228；Department of Plastic and Burns，the First Affiliated Hospital of Nanjing Medical University，Nanjing
210029，China
［Abstract］ 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）%，

［基金项目］ 苏州市吴江区科技局科研项目（WWK201615）
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通信作者（Corresponding author），E⁃mail：huyin282@sohu.com