miR-194-5p通过Wnt5a抑制肝星状细胞活化

doi:10.3969/j.issn.1000-1565.2023.02.010

摘要/Abstract

摘要： 为研究miR-194-5p对肝星状细胞(HSCs)活化的影响及作用机制,采用CCK-8、β-半乳糖苷酶染色、Transwell、划痕、集落形成实验和RT-qPCR、Western blot分别检测miR-194-5p 对LX-2细胞生物学特性、细胞活化标志物和靶基因表达的影响,并用生物信息学和双荧光素酶报告实验确定miR-194-5p靶基因. 结果显示:miR-194-5p抑制LX-2细胞增殖、细胞集落形成、迁移能力以及LX-2细胞中活化标志物的表达,促进LX-2细胞衰老;Wnt5a为miR-194-5p的直接靶基因.研究表明miR-194-5p通过靶定Wnt5a抑制LX-2细胞活化,miR-194-5p和Wnt5a可能是治疗肝纤维化新的靶点.

关键词: miR-194-5p, 肝纤维化, 肝星状细胞活化, 增殖, 迁移

Abstract: To investigate the effects and mechanism of miR-194-5p on the activation of hepatic stellate cells(HSCs), CCK-8, β-galactosidase staining, Transwell, wound healing, colony formation assays, RT-qPCR and Western blot were used to detect the effects of miR-194-5p on the cell biological properties, the expression of HSC activation markers and target gene, respectively. The candidate target genes of miR-194-5p were confirmed by the bioinformatics website and dual luciferase reporter assay. The results showed that the proliferation, cell colony formation, migration ability and HSC activation markers expression of LX-2 cells were inhibited by miR-194-5p; the senescence was promoted by miR-194-5p; and- DOI:10.3969/j.issn.1000-1565.2023.02.010miR-194-5p通过Wnt5a抑制肝星状细胞活化李亚琦¹,张荣花¹,刘雨潭¹,曹雨萌¹,熊亚南¹,章广玲²(1.华北理工大学基础医学院, 河北省慢性疾病重点实验室, 河北唐山 063200;2.华北理工大学临床医学院,河北省医工融合精准医疗重点实验室, 河北唐山 063200)摘要:为研究miR-194-5p对肝星状细胞(HSCs)活化的影响及作用机制,采用CCK-8、β-半乳糖苷酶染色、Transwell、划痕、集落形成实验和RT-qPCR、Western blot分别检测miR-194-5p 对LX-2细胞生物学特性、细胞活化标志物和靶基因表达的影响,并用生物信息学和双荧光素酶报告实验确定miR-194-5p靶基因. 结果显示:miR-194-5p抑制LX-2细胞增殖、细胞集落形成、迁移能力以及LX-2细胞中活化标志物的表达,促进LX-2细胞衰老;Wnt5a为miR-194-5p的直接靶基因.研究表明miR-194-5p通过靶定Wnt5a抑制LX-2细胞活化,miR-194-5p和Wnt5a可能是治疗肝纤维化新的靶点.关键词:miR-194-5p;肝纤维化;肝星状细胞活化;增殖;迁移中图分类号:R575.2 文献标志码:A 文章编号:1000-1565(2023)02-0179-09miR-194-5p inhibits activation of human hepatic stellate cells through Wnt5aLI Yaqi¹, ZHANG Ronghua¹, LIU Yutan¹, CAO Yumeng¹, XIONG Yanan¹, ZHANG Guangling²(1.Hebei Key Laboratory for Chronic Diseases, School of Basic Medicine, North China University of Science and Technology, Tangshan 063200, China; 2. Hebei Provincial Key Laboratory of Medical-Industrial Integration Precision Medicine, School of Clinical Medicine, North China University of Science and Technology, Tangshan 063200, China)Abstract: To investigate the effects and mechanism of miR-194-5p on the activation of hepatic stellate cells(HSCs), CCK-8, β-galactosidase staining, Transwell, wound healing, colony formation assays, RT-qPCR and Western blot were used to detect the effects of miR-194-5p on the cell biological properties, the expression of HSC activation markers and target gene, respectively. The candidate target genes of miR-194-5p were confirmed by the bioinformatics website and dual luciferase reporter assay. The results showed that the proliferation, cell colony formation, migration ability and HSC activation markers expression of LX-2 cells were inhibited by miR-194-5p; the senescence was promoted by miR-194-5p; and- 收稿日期:2022-06-07 基金项目:河北省自然科学基金资助项目(H2021209026);河北省人力资源和社会保障厅项目(C20210340);2020年政府资助临床医学优秀人才培养项目(冀财预付［2020］397号) 第一作者:李亚琦(1997—),女,河北邯郸人,华北理工大学在读硕士研究生,主要从事肝纤维化分子机制方向研究. E-mail:871528007@qq.com 通信作者:熊亚南(1978—),女,河北唐山人,华北理工大学副教授,主要从事肝纤维化分子机制方向研究.E-mail:xiongyanan@ncst.edu.cn章广玲(1972—),女,河北唐山人,华北理工大学教授,博士,主要从事肝纤维化分子机制方向研究.E-mail:zhanggl@ncst.edu.cn第2期李亚琦等:miR-194-5p通过Wnt5a抑制肝星状细胞活化河北大学学报(自然科学版) 第43卷Wnt5a was a target gene of miR-194-5p. Our findings provide evidence that miR-194-5p inhibits LX-2 cell activation by targeting Wnt5a. Therefore, miR-194-5p and Wnt5a may be possible novel therapeutic targets for hepatic fibrosis.

Key words: miR-194-5p, hepatic fibrosis, hepatic stellate cell activation, proliferation, migration

中图分类号:

R575.2

李亚琦,张荣花,刘雨潭,曹雨萌,熊亚南,章广玲. miR-194-5p通过Wnt5a抑制肝星状细胞活化[J]. 河北大学学报(自然科学版), 2023, 43(2): 179-187.

LI Yaqi, ZHANG Ronghua, LIU Yutan, CAO Yumeng, XIONG Yanan, ZHANG Guangling. miR-194-5p inhibits activation of human hepatic stellate cells through Wnt5a[J]. Journal of Hebei University(Natural Science Edition), 2023, 43(2): 179-187.

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