基于生物模型网络的肢体缺血再灌注损伤关键基因及免疫浸润机制研究  

Analysis of Key Genes and Immune Infiltration Mechanism of Limb Ischemia-Reperfusion Injury Based on Bioinformatics Model Network

作　　者：李彦丰 郝洋 石淇允 LI Yanfeng;HAO Yang;SHI Qiyun(Luoyang Orthopedic-Traumatological Hospital of Henan Province/Henan Provincial Orthopedic Hospital,Henan Zhengzhou 450000,China;Henan University of Chinese Medicine,Henan Zhengzhou 450000,China)

机构地区：[1]河南省洛阳正骨医院/河南省骨科医院,河南郑州450000 [2]河南中医药大学,河南郑州450000

出　　处：《中国医药导刊》2024年第9期917-928,共12页Chinese Journal of Medicinal Guide

基　　金：河南省医学科技攻关计划省部共建项目(SBGJ202103042);河南省中医药科学研究专项(2017ZY2121)。

摘　　要：目的:建立生物模型网络,分析肢体缺血再灌注损伤(IRI)关键基因及免疫浸润机制并实验验证。方法:通过GEO数据库获取IRI芯片。使用随机森林、LASSO回归和SVM算法筛选IRI相关特征基因。构建肢体IRI动物模型,用RT-qPCR实验验证关键基因mRNA相对表达量。用CIBERSORT算法预估IRI与筛选出的潜在生物标志物的生物信息学关联。结果:共获取差异基因169个,其中上调基因116个,下调基因53个。通过2种机器学习算法筛选到最相关的4个相关特征基因(WNT5A、PLCG、ITPR1、CAMK2A),RT-qPCR结果显示,与对照组相比,IRI组中WNT5A、PLCG、ITPR1、CAMK2A表达量上调(P<0.01)。免疫细胞浸润显示,IRI组中浆细胞、滤泡辅助性T细胞、初始世噬细胞、M2型巨噬细胞的浸润程度降低(P<0.05);M1型巨噬细胞、静止树实状细胞、活化肥大细胞的浸润程度增高(P<0.05)。静息状态肥大细胞与单核细胞(r=0.76)、M2型巨噬细胞与CD8+T细胞(r=0.75)、M2型巨噬细胞与活化肥大细胞(r=0.75)等呈现正相关;静息状态肥大细胞与M2型巨噬细胞(r=-0.88)、M2型巨噬细胞与M1型巨噬细胞(r=-0.86)、活化的自然杀伤细胞与初始B细胞(r=-0.75)等之间呈现负相关。WNT5A记忆B细胞、记忆CD4+T细胞呈负相关;PLCG与NK激活细胞呈正相关;CAMK2A与初始B细胞呈正相关;ITPR1与γδT细胞呈正相关。结论:WNT5A、PLCG、ITPR1、CAMK2A可能在肢体IRI发病过程中起重要作用,并与多种免疫细胞存在相关性,在防治肢体IRI的过程中,同时应重视免疫细胞作用。Objective:To establish a bioinformatics model to analyze key genes and immune infiltration mechanism of limb ischemia-reperfusion injury(IRI)and to experimentally validate the findings.Methods:IRI microarray data were obtained from the GEO data-base.Random forest method,LASSO regression,and SVM algorithms were employed to identify IRI-related feature genes.An animal model of limb IRI was constructed,and RT-qPCR was used to verify the relative mRNA expression levels of key genes.The CIBERSORT algorithm was utilized to estimate the association between IRI and the identified potential biomarkers in terms of bioinformatics.Results:A total of 169 differentially expressed genes(DEGs)were identified,with 116 upregulated and 53 downregulated.Four highly relevant feature genes(WNT5A,PLCG,ITPR1,CAMK2A)were screened using two machine learning algorithms.RT-qPCR results showed that the expression levels of WNT5A,PLCG,ITPR1,and CAMK2A significantly upregulated in the IRI group compared to the control group(P<0.01).Immune cell infiltration analysis revealed that plasma cells,T cells follicular helper,macrophages M0,and macrophages M2 significantly reduced in the IRI group(P<0.05),while macrophages M1,dendritic cells resting,and mast cells activated significantly increased(P<0.05).Strong positive correlations were observed between mast cells resting and monocytes(r=0.76),macrophages M2 and T cells CD8(r=0.75),and macrophages M2 and mast cells activated(r=0.75).Conversely,strong negative correlations were found between mast cells resting and macrophages M2(r=-0.88),macrophages M2 and macrophages M1(r=-0.86),and NK cells activated and B cells naive(r=-0.75).WNT5A showed a negative correlation with memory B cells and memory CD4+T cells.PLCG showed a positive correlation with NK activated cells.CAMK2A showed a positive correlation with naive B cells.ITPR1 showed a positive correla-tion withγδT cells.Conclusion:WNT5A,PLCG,ITPR1,and CAMK2A may play important roles in the pathogenesis of limb IRI and are associated with various immu

关 键 词：生物模型网络 肢体缺血再灌注损伤 免疫细胞浸润 

分 类 号：R743.3[医药卫生—神经病学与精神病学]