大鼠骨折愈合过程中时序性相关靶基因的筛选及生物信息学分析  

Screening and Bioinformatics Analysis of Time-related Key Genes During Fracture Healing in Rats

作　　者：张晶晶 洪莉[1] 王治 ZHANG Jingjing;HONG Li;WANG Zhi(Department of Gynecology and Obestetrics of Peoples Hospital,Wuhan University,Wuhan 430060,China;Xiaogan Public Security Bureau,Xiaogan 432000,Hubei,China)

机构地区：[1]武汉大学人民医院妇产科,武汉430060 [2]孝感市公安局,湖北孝感432000

出　　处：《刑事技术》2025年第1期48-55,共8页Forensic Science and Technology

基　　金：国家重点研发计划(2021YFC2701303);国家自然科学基金(81971364);湖北省重点研发项目(2022BCA045);湖北省领军人才(201947);湖北省自然科学基金(2022CFB124)。

摘　　要：本文应用生物信息学方法探讨外力导致骨折后损伤修复过程中相关靶基因的表达,初步筛选出不同时间节点的关键基因,为骨折损伤时间推断提供理论依据。从基因表达数据库下载基因芯片数据集GSE594,使用R软件分别筛选伤后3 d组、1周组、2周组、4周组、6周组与正常对照组比较的差异基因,进行GO和KEGG富集分析。采用Cytoscape软件将差异基因导入进行蛋白网络互作分析,利用3种算法筛选排名前15的基因,韦恩图取交集确定关键基因。采用CIBERSORT反卷法分析骨折后22类免疫细胞的含量及比例。结果发现,与正常对照组相较,大鼠骨折后3 d组关键基因为:IL6、SRC、CASP3、TGFB1、PRKACA、HAPA5;1周组关键基因为:IL6、MMP9、DCN、ACAN、SDC1;2周组关键基因为:IL6、MMP9、COL3A1、ACAN、COL2A1、SDC1;4周组关键基因为:MMP9、TIMP1、SPARC、SPP1、APOE、SDC1、CTSK;6周组关键基因为:TIMP1、COL2A1、ACAN、COL10A1、MMP13、SERPINH1、COL5A1、PCOLCE、APOE、CTSK、THY1、FGF5、GPX3。GO功能分析显示关键基因与伤口愈合、骨化、结缔组织发育、软骨发育有关,KEGG分析显示关键基因与P13K-Akt通路、HIF-1信号通路、焦点粘附通路、成骨细胞分化通路有关。关键基因在骨折愈合过程不同阶段发挥特定表达作用,与损伤时间密切相关。对免疫细胞的浸润矩阵分析结果表明,巨噬细胞在骨折后显著增高,且持续时间较长,从免疫细胞层面为进一步了解骨折愈合过程、推断骨折后损伤时间提供了依据。In this paper,bioinformatics methods were used to investigate the expression of related genes in the process of injury repair after fracture induced by external force,to provide a theoretical basis for the estimation of fracture injury time.Data set GSE594 was downloaded from the GEO,and the differential genes were screened by R software in 3-day group,1-week group,2-week group,4-week group and 6-week group,respectively.Enrichment analysis of these differential genes was performed using GO and KEGG databases.Cytoscape software was used to introduce differential genes into protein network interaction analysis,where three algorithms were employed to screen the top 15 genes.Venn diagram intersection was used to identify key genes.The content and proportion of 22 kinds of immune cells in control group and injury group after fracture were analyzed using CIBERSORT deconvolution method.Compared with the normal control group,the key genes in the group 3 days after fracture were:IL6、SRC、CASP3、TGFB1、PRKACA、HAPA5;the key genes in the 1-week group were:IL6、MMP9、DCN、ACAN、SDC1;the key genes in the 2-week group were:IL6、MMP9、COL3A1、ACAN、COL2A1、SDC1;the key genes in 4-week group were:MMP9、TIMP1、SPARC、SPP1、APOE、SDC1、CTSK;the key genes in 6-week group were:TIMP1、COL2A1、ACAN、COL10A1、MMP13、SERPINH1、COL5A1、PCOLCE、APOE、CTSK、THY1、FGF5、GPX3.GO functional analysis of the key geneshowed that it was associated with wound healing,ossification,connective tissue development and cartilage development.KEGG analysis showed that P13K-Akt pathway,HIF-1 signaling pathway,focal adhesion pathway and osteoblast differentiation pathway were involved.The key genes play a specific role in the different stages of fracture healing and are related to the time of fracture injury.The results of infiltration matrix analysis of immune cells showed that macrophages increased significantly after fracture and lasted for a long time,which provides a basis for further study of fracture healing process a

关 键 词：法医学 关键基因 生物信息学 骨折损伤时间 

分 类 号：DF795.1[医药卫生—法医学]