2023年河北大学学报(自然科学版)2023第43卷第2期JournalofHebeiUniversity(NaturalScienceEdition)Vol.43No.2DOI:10.3969/j.issn.10001565.2023.02.010miR-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、Westernblot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:10001565(2023)02017909miR-194-5pinhibitsactivationofhumanhepaticstellatecellsthroughWnt5aLIYaqi1,ZHANGRonghua1,LIUYutan1,CAOYumeng1,XIONGYanan1,ZHANGGuangling2(1.HebeiKeyLaboratoryforChronicDiseases,SchoolofBasicMedicine,NorthChinaUniversityofScienceandTechnology,Tangshan063200,China;2.HebeiProvincialKeyLaboratoryofMedical-IndustrialIntegrationPrecisionMedicine,SchoolofClinicalMedicine,NorthChinaUniversityofScienceandTechnology,Tangshan063200,China)Abstract:ToinvestigatetheeffectsandmechanismofmiR-194-5pontheactivationofhepaticstellatecells(HSCs),CCK-8,β-galactosidasestaining,Transwell,woundhealing,colonyformationassays,RT-qPCRandWesternblotwereusedtodetecttheeffectsofmiR-194-5ponthecellbiologicalproperties,theexpressionofHSCactivationmarkersandtargetgene,respectively.ThecandidatetargetgenesofmiR-194-5pwereconfirmedbythebioinformaticswebsiteanddualluciferasereporterassay.Theresultsshowedthattheproliferation,cellcolonyformation,migrationabilityandHSCactivationmarkersexpressionofLX-2cellswereinhibitedbymiR-194-5p;thesenescencewaspr...
