CAJ | 학술논문

目的探讨对甾体类激素导致股骨头坏死的分子机制.方法使用Ⅰ型胶原酶和胰蛋白酶消化法分离人股骨头松质骨内的微血管内皮细胞,通过血管假性血友病因子、血小板-内皮细胞黏附分子-1、血管内皮钙黏蛋白和血管细胞黏附分子-1等血管内皮细胞标志物抗体进行免疫荧光鉴定,用双标的lncRNAs+mRNAs表达谱芯片和单标microRNAs芯片进行差异性转录本检测,通过生物信息学软件并参考数据库信息,分析出lncRNAs-microRNAs-mRNAs的空间位置和表达水平的相关性.结果体外培养的股骨头内微血管内皮细胞在甾体类激素损伤情况下出现mRNAs、microRNAs(其中miR-339上调,miR-100、miR-222、miR-23、miR-933下调)和lncRNAs(其中73种表达上调,166种表达下调)等分子特异性的变化,利用靶基因预测等生物信息学工具,构建出了转录本模块间的共表达调控网络模型.结论甾体类激素导致骨内微血管内皮细胞miR-100、miR-222、miR-933、miR-339和miR-23等转录本发生明显变化,它们直接攻击过氧化氢酶、成纤维细胞生长因子和神经生长因子等关键酶和细胞因子,对松质骨内的骨组织和循环系统产生重要影响.
목적 탐토대치체류격소도치고골두배사적분자궤제.방법 사용Ⅰ형효원매화이단백매소화법분리인고골두송질골내적미혈관내피세포,통과혈관가성혈우병인자、혈소판-내피세포점부분자-1、혈관내피개점단백화혈관세포점부분자-1등혈관내피세포표지물항체진행면역형광감정,용쌍표적lncRNAs+mRNAs표체보심편화단표microRNAs심편진행차이성전록본검측,통과생물신식학연건병삼고수거고신식,분석출lncRNAs-microRNAs-mRNAs적공간위치화표체수평적상관성.결과 체외배양적고골두내미혈관내피세포재치체류격소손상정황하출현mRNAs、microRNAs(기중miR-339상조,miR-100、miR-222、miR-23、miR-933하조)화lncRNAs(기중73충표체상조,166충표체하조)등분자특이성적변화,이용파기인예측등생물신식학공구,구건출료전록본모괴간적공표체조공망락모형.결론 치체류격소도치골내미혈관내피세포miR-100、miR-222、miR-933、miR-339화miR-23등전록본발생명현변화,타문직접공격과양화경매、성섬유세포생장인자화신경생장인자등관건매화세포인자,대송질골내적골조직화순배계통산생중요영향.
Objective To explore the differential expression profiles and transcriptomes of human bone microvascular endothelial cells via microarray and elucidate the molecular mechanisms underlying steroids-induced osteonecrosis.Methods Bone microvascular endothelial cells in cancellous bone of femoral head were harvested by type Ⅰ collagenase and trypsin digestion and identified by immunofluorescent labeling with primary antibodies specific for vascular endothelial cells' biomarkers such as yon Willebrand factor,platelet endothelial cell adhesion molecule 1,vascular endothelial cadherin and vascular cell adhesion molecule 1.Their differential expression profiles and transcriptomes were tested by double-labeled lncRNAs + mRNAs microarrays and single-labeledd microRNAs microarrays.The data were analyzed by bioinformatical software with database checking.The authors spatial correlations and expression relevance were established among lncRNAs,microRNAs and mRNAs.Results Steroids incurred specific changes in mRNAs,microRNAs and lncRNAs transcribed in human bone microvascular endothelial cells of femoral head in vitro cultured.Thus a model of coexpression network was constructed among transcript modules with bioinformatical tools such as target gene prediction.Conclusion Steroids induced significant changes in transcripts of bone microvascular endothelial cells such as miR-100,miR-222,miR-933,miR-339 and miR-23.They exert considerable influences on osseous tissues and circulatory systems in cancellous bone by targeting cytokines and enzymes such catalase,fibroblast growth factor and nerve growth factor.Through further explorations of these transcripts,we can not only elucidate the underlying pathogenesis,but also discover specific biomarkers and therapeutic targets for corresponding diseases.