miRNAs对人皮肤成纤维细胞光老化的作用及其相关机制

发布时间：2018-05-11 00:00

本文选题：miRNAs + 皮肤老化　；参考：《中国老年学杂志》2017年02期

【摘要】：目的探讨miR-146a在紫外线诱导人皮肤成纤维细胞光老化中的作用。方法 10 J/cm~2UVA连续照射成纤维细胞2 w建立光老化模型,分别于首日、3、7、14 d提取总RNA,实时定量PCR检测miR-146a表达量;慢性病毒转染上调miR-146a表达(miR-146a过表达组),荧光显微镜观察转染效率,并验证细胞内miR-146a表达量;未经处理的成纤维细胞作为空白对照组;UVA+miR-146a组为miR-146a过表达后再用UVA照射。MTT法检测四组细胞增殖情况;实时定量PCR检测四组细胞内老化相关基因mRNA表达情况;Western印迹法检测四组细胞中Smad4蛋白表达情况。结果 UVA照射第3、7、14天UVA照射组miR-146a表达量持续降低,且明显低于同期空白对照组(P0.05)。慢性病毒载体转染后,miR-146a过表达组第7、14天miR-146a表达量明显高于同期空白对照组(P0.05)。UVA照射组、UVA+miR-146a组A值明显低于空白对照组、miR-146a过表达组(P0.01);miR-146a过表达组A值与空白对照组之间无统计学差异(P0.05);UVA+miR-146a组A值明显高于UVA照射组(P0.01)。UVA照射组、UVA+miR-146a组p53、p21、p16 mRNA表达量均明显高于空白对照组、miR-146a过表达组(P0.01);miR-146a过表达组p53、p21、p16mRNA表达量与空白对照组之间无统计学差异(P0.05);UVA+miR-146a组p53、p21、p16 mRNA表达量明显低于UVA照射组(P0.01)。UVA照射组、UVA+miR-146a组细胞内Smad4蛋白表达量明显高于空白对照组、miR-146a过表达组(P0.01);miR-146a过表达组与空白对照组Smad4蛋白表达量之间无统计学意义(P0.05);UVA+miR-146a组Smad4蛋白表达量明显低于UVA照射组(P0.01)。结论 UVA诱导光老化人皮肤成纤维细胞中miR-146a表达被抑制,上调其表达可抑制Smad4表达,促进光老化细胞增殖,起到抗老化作用。
[Abstract]:Objective to investigate the role of miR-146a in ultraviolet induced photoaging of human skin fibroblasts. Methods the model of photoaging was established by 10 J/cm~2UVA continuous irradiation of fibroblasts for 2 weeks. The total RNAs were extracted on the first day, and the expression of miR-146a was measured by real-time quantitative PCR, and the expression of miR-146a was up-regulated by chronic virus transfection, and the transfection efficiency was observed by fluorescence microscope. The untreated fibroblasts were used as the blank control group for miR-146a overexpression and then the proliferation of the four groups was detected by UVA irradiation. Real-time quantitative PCR was used to detect the mRNA expression of aging related genes in four groups of cells. Western blot was used to detect the expression of Smad4 protein in the four groups. Results the expression of miR-146a in the UVA group was significantly lower than that in the control group on the 14th day of UVA irradiation, and was significantly lower than that in the control group (P 0.05). After transfection of chronic virus vector, the expression of miR-146a in the overexpression group of miR-146a was significantly higher than that in the control group (P0.05N. UVA irradiation group) on the 14th day after transfection, and the A value in the miR-146a group was significantly lower than that in the blank control group and the blank control group in which the expression of miR-146a overexpression group was significantly lower than that in the control group and the blank control group. There was no significant difference in the expression of p53p21, p16 mRNA between the UVA irradiation group and the UVA miR-146a group, and the expression of p53p21 + p16 mRNA in the UVA miR-146a group was significantly higher than that in the control group (P0.01 / 146a overexpression group). There was no significant difference in the expression of p53p21p16mRNA between the UVA irradiation group and the blank control group (P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA / P0.05UVA The expression of Smad4 protein in the miR-146a group was significantly lower than that in the UVA irradiation group. The expression of Smad4 protein in the miR-146a group was significantly higher than that in the blank control group. There was no significant difference in the expression of Smad4 protein between the control group and the blank control group. The expression of Smad4 protein in UVA miR-146a group was significantly lower than that in UVA irradiation group. Conclusion UVA induces the inhibition of miR-146a expression in human skin fibroblasts induced by photoaging, and up-regulation of miR-146a expression can inhibit the expression of Smad4, promote the proliferation of photoaged cells, and play an anti-aging role.
【作者单位】：四川省医学科学院四川省人民医院皮肤性病研究所;
【基金】：国家“973”计划项目(No.2011CB707805) 国家临床重点专科建设项目(〔2014〕1081) 四川省国际科技合作与交流计划项目(No.2015KW-041) 四川省科技厅项目(No.2013JY0194)
【分类号】：R758.1

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