【摘要】 目的:研究慢性光损伤皮肤细胞中对cathepsin D具有调控作用的长链非编码RNA(LncRNAs),并探讨其在皮肤光老化机制中的作用。方法:取包皮组织分离培养成纤维细胞,连续UVA照射诱导成纤维细胞慢性光损伤,与空白对照组细胞比较,通过CCK8、β-半乳糖苷酶染色及细胞凋亡率检测验证建模。LncRNAs差异表达检测及功能注释分析慢性光损伤细胞中可调控cathpeisn D的LncRNA;随后用miRanda和TargetScan检测可靶向结合LncRNA-cathpeisn D的miRNA。结果:UVA照射组细胞活性明显低于对照组[(60.02±5.13)%比(89.06±4.21)%,t=2.31,P<0.05];细胞老化率高于对照组[(85.24±4.21)%比(8.16±1.62)%,t=3.22,P<0.05];细胞凋亡率高于对照组[(26.04±7.42)%比(11.31±4.95)%,t=3.54,P<0.05]。LncRNAs差异表达检测及功能注释分析发现可调控cathepsin D且存在差异表达的LncRNA共有25个,其中表达差异最为明显的是Lnc-CTSD、Lnc-TNNI1和Lnc-RP11-706015.1.1-3。miRanda和TargetScan分析发现在慢性紫外线损伤细胞中,LncRNA-CTSD、Lnc-TNNI以及Lnc-RP11均通过与miR4298靶向结合发挥下游调控作用。结论:反复UVA照射可上调cathepsin D的调控LncRNAs, Lnc-CTSD、Lnc-TNNI1和Lnc-RP11-706015.1.1-3均通过与miR4298靶向结合发挥作用,可能通过microRNA-LncRNA-cathepsin D机制参与皮肤光老化发生。更多还原

【Abstract】 Objective:To investigate the role of LncRNAs that regulate cathepsin D in chronic light-damaged skin cells and explore their role in the mechanism of skin photoaging.Methods: Cultured human dermal fibroblasts were induced to a photoaging cells model by repetitive UVA radiation(UVA radiation group),which was evaluated by CCK8 assay, β-galactosidase staining and flow cytometry detection of apoptosis rate. High-throughput sequencing was used to detect LncRNA expression profiles. Functional annotation analysis was preformed to identify the LncRNAs that could regulate cathpeisn D. miRanda and TargetScan were preformed to analyze the miRNA that can target binding to LncRNA-cathpeisn D. Results:Compared with cells in the control group, the proliferative activity of cells in the UVA radiation group significantly decreased [(60.02±5.13)% vs(89.06±4.21)%, t=2.31,P<0.05],while the apoptosis rate of cells in the UVA radiation group significantly increased [(26.04±7.42)% vs(11.31±4.95)%,t=3.54,P<0.05], and the percentage of β-galactosidase of cells in the UVA radiation group significantly increased [(85.24± 4.21)% vs(8.16±1.62)%, t=3.22,P<0.05].Differential expression of LncRNAs was detected via high throughput sequencing technique. 25 differentially expressed LncRNAs were related to regulate cathepsin D including Lnc-CTSD, Lnc-TNNI1 and Lnc-RP11-706015.1.1-3 targeting to miR4298.Conclusion: Repeated UVA irradiation could up-regulate the expression of LncRNAs which regulate cathepsin D and all target to miR4298. The results in this study indicate that microRNA-LncRNA-cathepsin D may play an important role in skin photoaging.更多还原