基于ceRNA网络探讨lncRNA NEAT1调控肝纤维化的分子机制
广西中医药大学, 赛恩斯新医药学院医学系, 广西南宁 530222
| 摘 要： | 目的：借助生物信息学和实验验证分析lncRNA NEAT1调控肝纤维化的分子机制，希冀为肝纤维化的防治和药物研发提供新的思路。方法：借助Starbase数据库筛选lncRNA NEAT1靶向miRNA；使用miRDB数据库筛选miRNA靶向mRNA；通过OMIM、CTD、GeneCards数据库获取肝纤维化相关靶点；将lncRNA NEAT1调控靶点与肝纤维化靶点进行交互处理，对关键靶点进行富集分析。用LPS活化肝星状细胞（HSC）将其分为空白组和模型组，用siNEAT1和阴性对照对HSC进行转染，将其分为siNEAT1 NC+LPS、siNEAT1+LPS，MTT法检测HSC增殖情况，流式细胞术检测HSC的凋亡率；使用分子生物学方法检测α-SMA和CollagenⅠ的表达。结果：lncRNA NEAT1靶向miRNA分子26个，miRNA靶向mRNA564个，肝纤维化相关靶点617个，lncRNA NEAT1调控肝纤维化关键靶点20个，lncRNA NEAT1参与肝纤维化可能与其调控细胞衰老关系密切。HSC活化时NEAT1表达显著增加，沉默NEAT1后HSC细胞增殖率显著下降，且凋亡率显著增加，α-SMA和CollagenⅠ表达也显著下降。结论：NEAT1通过调控HSC增殖和凋亡参与肝纤维化的发生发展，可能与其参与的ceRNA网络调控细胞衰老关系密切。 |
| 关 键 词： | lncRNA NEAT1; ceRNA; 凋亡; 衰老; 肝星状细胞; 肝纤维化 |
| DOI： | 10.57237/j.life.2023.01.003 |
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收稿日期: 2023-02-24
接受日期: 2023-04-12
在线出版日期: 2023-04-25
Exploring the Molecular Mechanism of lncRNA NEAT1 Regulation of Liver Fibrosis Based on ceRNA Network
Department of Medicine, Faculty of Chinese Medicine Science, Guangxi University of Chinese Medicine, Nanning 530222, China
| Abstract: | Objective: To analyze the molecular mechanism of lncRNA NEAT1 regulation of liver fibrosis with the help of bioinformatics and experimental validation, hoping to provide new ideas for the prevention and treatment of liver fibrosis and drug development. Methods: Screening lncRNA NEAT1 target miRNAs with the help of Starbase database; screening miRNA target mRNAs using miRDB database; obtaining liver fibrosis-related targets through OMIM, CTD and GeneCards databases; interacting lncRNA NEAT1 regulatory targets with liver fibrosis targets, and performing key target Enrichment analysis was performed. Hepatic stellate cells (HSC) were activated with LPS and divided into blank and model groups. HSC were transfected with siNEAT1 and negative control and divided into siNEAT1 NC+LPS and siNEAT1+LPS. MTT method was used to detect the proliferation of HSC and flow cytometry was used to detect the apoptosis rate of HSC; molecular biology was used to detect α-SMA and Collagen I expression. Results: lncRNA NEAT1 targeted 26 miRNA molecules, miRNA targeted 564 mRNAs, 617 liver fibrosis-related targets, lncRNA NEAT1 regulated 20 key targets of liver fibrosis, lncRNA NEAT1 involvement in liver fibrosis may be closely related to its regulation of cellular senescence. lncRNA NEAT1 expression increased significantly upon HSC activation. The proliferation rate of HSC cells was significantly decreased and apoptosis rate was significantly increased after NEAT1 silencing, and the expression of α-SMA and Collagen I was also significantly decreased. Conclusion: NEAT1 is involved in the development of liver fibrosis by regulating HSC proliferation and apoptosis, and may be closely related to its involvement in the ceRNA network that regulates cellular senescence. |
| Keywords: | lncRNANEAT1; ceRNA; Apoptosis; Senescence; Hepatic Stellate Cells; Hepatic Fibrosis |
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