基于高通量测序的激素性股骨头坏死关键基因的筛选与分析  

Screening and analysis of key genes for steroid-induced osteonecrosis of the femoral head based on high-throughput sequencing

作　　者：高航 杨治[2] 张铭[3] 胡守业 Gao Hang;Yang Zhi;Zhang Ming;Hu Shouye(The Second Clinical Medical College of Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西中医药大学第二临床医学院,咸阳712046 [2]陕西省西安交通大学附属红会医院骨坏死与关节重建病区 [3]陕西省西安交通大学附属红会医院全科医学科

出　　处：《山西医药杂志》2023年第13期963-966,共4页Shanxi Medical Journal

基　　金：陕西省科学技术厅自然科学青年基础项目(2020JQ-964);陕西省西安市科技局项目(2019115013YX00-5SF038(9))。

摘　　要：目的探索激素性股骨头坏死(SONFH)基因表达上的差异,筛选出关键致病基因,为SONFH诊疗提供新的靶点。方法选取我院2020年12月至2021年6月SONFH患者和股骨颈骨折患者18例作为研究对象,SONFH患者设为SONFH组(6例),股骨颈骨折患者设为对照组(12例),通过高通量测序筛选出2组患者骨组织中差异表达基因(DEGs)以及差异MicroRNAs(miRNAs),利用R软件Clusterprofiler包对DEGs进行富集分析。结合P值选取前30位DEGs进行文献调研,筛选关键致病基因,并通过实时荧光定量聚合酶联反应(qRT-PCR)验证其表达。使用Mir SNPInTarget、miRWalk和miRDB数据库预测关键致病基因靶向的mi RNAs。结果共得到735个DEGs(482个上调,253个下调)和111个差异miRNAs(34个上调,77个下调)。P值较低的前30位DEGs中,HMOX1在SONFH组中呈下调表达。qRT-PCR结果显示HMOX1在SONFH组表达低于对照组,差异有统计学意义(P<0.05)。HMOX1靶向的2个关键miRNAs为hsa-miR-21-5p、hsa-mi R-148a-3p。结论HMOX1可能成为SONFH潜在的关键致病基因之一,参与了SONFH的发生与发展,为SONFH诊疗提供新靶点。Objective Explore the differences in the expression of steroid-induced osteonecrosis of the femoral head(SONFH)gene,and select out the key pathogenic genes to provide new targets for the diagnosis and treatment of SONFH.Methods Eighteen SONFH patients and femoral neck fracture patients from December 2020 to June 2021 were selected as study subjects,SONFH patients were set as the SONFH group(6 patients)and femoral neck fracture patients were set as the control group(12 patients),Differentially expressed genes(DEGs)and differential MicroRNAs(miRNAs)in the two groups were selected by high-throughput sequencing,and enrichment analysis of DEGs was performed using the R software Clusterprofiler package.The top 30 DEGs were selected for literature investigation,key pathogenic genes were screened,and their expression was verified by real-time PCR(qRT-PCR).To of miRNAs targeted by key pathogenic genes using the MirSNPInTarget,miRWalk and miRDB databases.Results A total of 735 DEGs(482 up,253 downregulated)and 111 differential miRNAs(34 up,77 down)were obtained.Among the top 30 DEGs with lower P values,HMOX 1 was down-regulated in the SONFH group.The qRT-PCR results showed that HMOX 1 expression was lower in the SONFH group than in the control group,with a statistically significant difference(P<0.05).The two key miRNAs targeted by HMOX 1 were hsa-miR-21-5p and hsa-miR-148a-3p.Conclusion HMOX1 may become one of the potential key pathogenic genes of SONFH,participate in the occurrence and development of SONFH,and provide new targets for the diagnosis and treatment of SONFH.

关 键 词：股骨头坏死 激素性 差异表达 基因 微RNAS 血红素加氧酶1 

分 类 号：R681.8[医药卫生—骨科学]