has-miR-100-5p靶向含7A的锌指和BTB结构域是绝经后骨质疏松的潜在靶标  

Has-miR-100-5p targeting zinc finger and BTB domain containing 7A is a potential target for postmenopausal osteoporosis

作　　者：麦合木提江·穆海麦提 买买提沙吾提阿吉·买买提 唐军伟[1] 张玉新[1] Maihemutijiang·Muhaimaiti;Maimaitishawutiaji·Maimaiti;Tang Junwei;Zhang Yuxin(Department of Spinal Orthopedics,the First People’s Hospital of Kashgar,Kashgar 844099,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]喀什地区第一人民医院脊柱骨科,新疆维吾尔自治区喀什市844099

出　　处：《中国组织工程研究》2023年第32期5168-5172,共5页Chinese Journal of Tissue Engineering Research

基　　金：喀什地区科技计划项目(KS2020008),项目负责人:麦合木提江·穆海麦提。

摘　　要：背景:miRNAs在骨质疏松症的发病机制中起重要作用,因此可以成为重要的治疗靶点。其中,靶向miR-100-5p调控轴促进成骨细胞分化是骨质疏松的潜在治疗靶标。目的:探讨has-miR-100-5p和has-miR-101-3p在绝经后骨质疏松中的靶向调控作用。方法:比较GSE56814和GSE56815数据中高骨矿物质密度和低骨矿物质密度之间的差异表达基因,并进行富集分析。预测差异表达基因中has-miR-100-5p和has-miR-101-3p的靶向调控的基因。随后,收集40例绝经后骨质疏松患者和40例健康对照者的外周血样本,采用qRT-PCR法检测has-miR-100-5p和has-miR-101-3p以及靶标基因的差异表达。最后,通过双荧光素酶报告基因检测has-miR-100-5p对靶标基因3’UTR的结合作用。结果与结论:①富集分析显示,高骨矿物质密度和低骨矿物质密度之间鉴定到81个共同差异表达基因,显著富集于免疫、核因子κB复合物、肿瘤坏死因子信号通路等;②miRTargetbase数据库和miRTarget数据库预测显示,has-miR-100-5p和has-miR-101-3p均靶向调控含7A的锌指和BTB结构域(zinc finger and BTB domain containing,ZBTB7A);③qRT-PCR检测显示,与健康对照组比较,骨质疏松组has-miR-100-5p mRNA表达上调(P<0.001),has-miR-101-3p和ZBTB7A mRNA表达下调(P<0.001);双荧光素酶报告系统显示,野生型ZBTB7A-3’UTR共转染has-miR-100-5p后,293T细胞的荧光活性下调;④结果显示,has-miR-100-5p靶向ZBTB7A可能是绝经后骨质疏松症的潜在靶标。BACKGROUND:miRNAs play an important role in the pathogenesis of osteoporosis,which therefore can be important therapeutic targets.Among them,targeting the miR-100-5p regulatory axis to promote osteoblast differentiation is a potential therapeutic target for osteoporosis.OBJECTIVE:To explore the targeted regulatory roles of has-miR-100-5p and has-miR-101-3p in postmenopausal osteoporosis.METHODS:Differentially expressed genes between high and low bone mineral density in the GSE56814 and GSE56815 datasets were identified and subjected to enrichment analyses.Target genes of has-miR-100-5p and has-miR-101-3p among the differentially expressed genes were predicted.Peripheral blood samples from 40 patients with postmenopausal osteoporosis and 40 healthy controls were selected and qRT-PCR experiments were performed to detect the differential expression of has-miR-100-5p and has-miR-101-3p as well as target genes.Dual luciferase reporter assay was used to measure the targeting effect of miRNAs to the 3'UTR of target genes.RESULTS AND CONCLUSION:A total of 81 common differentially expressed genes were identified between high and low bone mineral density,mainly significantly enriched in immunity,nuclear factor-κB compound,and tumor necrosis factor signaling pathway.Has-miR-100-5p and has-miR-101-3p were targeted to regulate zinc finger and BTB domain containing 7A(ZBTB7A)based on miRTargetbase database and miRTarget database predictions.Has-miR-100-5p was up-regulated(P<0.001)and has-miR-101-3p and ZBTB7A were down-regulated(P<0.001)in postmenopausal osteoporosis patients compared with the healthy controls.The fluorescence activity of 293T cells was down-regulated after co-transfection of has-miR-100-5p with wild-type ZBTB7A-3’UTR.To conclude,has-miR-100-5p targeting ZBTB7A may be a potential target for postmenopausal osteoporosis.

关 键 词：绝经后骨质疏松 has-miR-100-5p has-miR-101-3p ZBTB7A 骨矿物质密度 

分 类 号：R459.9[医药卫生—治疗学] R-331[医药卫生—临床医学] R589.5