基金项目:新疆维吾尔自治区自然科学基金项目资助(2020D01C223)作者单位:830000乌鲁木齐,新疆医科大学第五附属医院骨科中心(张峥、崔泳、丁路、王武、黄涛);830054乌鲁木齐,新疆医科大学第一附属医院骨肿瘤外科(王翀)通信作者:崔泳,电子信箱:14753634@qq.comlncRNAHOTAIR/miR-17-5p/Smad7通路调控激素性股骨头坏死的机制研究张峥崔泳王翀丁路王武黄涛摘要目的研究lncRNAHOTAIR对激素性股骨头坏死(steroidinducednecrosisoffemoralhead,SONFH)的影响。方法收集激素性股骨头坏死患者骨髓组织25例,正常对照组为25例股骨颈骨折患者。培养人骨髓间充质干细胞(hBMSCs),用地塞米松(DEX)培养,CCK-8测定细胞增殖。用HOTAIR过表达载体或miR-17-5p模拟物转染细胞。hBMSCs分为hBMSCs组、hBMSCs+成骨诱导组、hBMSCs+成骨诱导+空载组、hBMSCs+成骨诱导+过表达HOTAIR组、hBMSCs+成脂诱导组、hBMSCs+成脂诱导+空载组和hBMSCs+成脂诱导+过表达HOTAIR组。RT-qPCR测HOTAIR、miR-17-5p、Smad7、RUNX2、RRAR-γ的mRNA表达。Westernblot法检测Smad7蛋白的表达。双荧光素酶报告基因测法确认HOTAIR和miR-17-5p间的结合。碱性磷酸酶(ALP)活力检测试剂盒与茜素红染色评估细胞的成骨分化;油红O染色评估细胞的成脂分化。结果HOTAIR和Smad7在激素性股骨头坏死组织中表达上调,而miR-17-5p表达下调(P均<0.05)。双荧光素酶报告实验证实hBMSCs中HOTAIR的直接结合靶点是miR-17-5p,而Smad7是miR-17-5p的靶基因。HOTAIR的过表达诱导了hBMSCs的成脂分化和RRAR-γ表达,并抑制了成骨分化和RUNX2表达(P均<0.05)。结论过表达lncRNAHOTAIR直接靶向miR-17-5p并抑制hBMSCs的成骨分化,还能够促进成脂分化和Smad7的表达。关键词类固醇股骨头坏死miR-17-5p成骨分化成脂分化中图分类号R681.8;R274.9文献标识码ADOI10.11969/j.issn.1673-548X.2023.02.012MechanismofLncRNAHOTAIR/miR-17-5p/Smad7PathwayRegulatingSteroid-inducedFemoralHeadNecrosis.ZHANGZheng,CUIYong,WANGChong,etal.OrthopedicsCenter,FifthAffiliatedHospitalofXinjiangMedicalUniversity,Xinjiang830000,ChinaAbstractObjectiveToinvestigatetheeffectofLncRNAHOTAIRonsteroid-inducednecrosisfemoralhead(SONFH).Meth-odsBonemarrowtissuesof25patientswithSONFHwerecollected,and25patientswithfemoralneckfractureswereincludedinthecontrolgroup.Humanbonemarrowmesenchymalstemcells(hBMSCs)wereculturedwithDexamethason...
