基于高通量测序的风湿性心脏病瓣膜组织mRNA差异分析

Analysis of differentially expressed mRNAs in valve tissue of rheumatic heart disease based on next-generation sequencing

目的:基于高通量测序(NGS)技术筛选风湿性心脏病(RHD)与非RHD患者的瓣膜组织中的差异mRNA。方法:选取2020—2021年在广西医科大学第一附属医院行二尖瓣置换术的RHD患者(RHD组)和非RHD患者(Con组),取瓣膜组织进行基因NGS,对显著差异基因进行基因功能(GO)富集分析和京都基因与基因百科全书(KEGG)通路分析,然后通过蛋白相互作用(PPI)网络筛选关键基因,通过实时荧光定量PCR(RT-qPCR)验证关键基因表达。结果:两组瓣膜组织共4498个差异表达基因,其中2315个上调,2183个下调,显著差异表达有530个,筛选出FN1、CD44、VCAM-13个关键基因。与Con组比较,FN1、CD44、VCAM-1在RHD组瓣膜组织中的表达均上调(P<0.05)。GO分析结果显示,主要富集的生物学过程(BP)为细胞外基质(ECM)及结构的组装和分解,主要的细胞成分(CC)为细胞外基质,显著的分子功能(MF)为钙黏着蛋白结合和肌动蛋白结合。KEGG通路显著富集于细胞基质黏着、白细胞跨内皮转移、ECM受体相互作用等信号通路中。结论:NGS筛查出RHD患者与非RHD患者瓣膜组织中差异表达基因FN1、CD44、VCAM-1,可能在ECM受体相互作用和白细胞跨内皮转移等信号通路中发挥重要作用。

Objective:To screen differentially expressed mRNAs in the valve tissues of patients with and without rheumatic heart disease(RHD)based on next-generation sequencing(NGS)technology.Methods:Patients with RHD(RHD group)and non-RHD patients(Con group)who underwent mitral valve replacement at the First Affiliated Hospital of Guangxi Medical University from 2020 to 2021 were enrolled,and valve tissues were subjected to gene NGS.Gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed for significant differentially expressed genes,and key genes were screened by protein-protein interaction(PPI)network.Key gene expression was verified by real-time quantitative PCR(RT-qPCR).Results:There were 4,498 differentially expressed genes in the valve tissues of the two groups,of which 2,315 were up-regulated,2,183 were down-regulated,and 530 were significantly differentially expressed,and three key genes,FN1,CD44,and VCAM-1,were screened.Compared with the Con group,the expressions of FN1,CD44,and VCAM-1 were all upregulated in the valve tissue of the RHD group(P<0.05).GO analysis showed that the major enriched biological processes(BP)were assembly and disassembly of extracellular matrix(ECM)and structures,the major cellular component(CC)was extracellular matrix,and the significant molecular functions(MF)were cadherins binding and actin binding.The KEGG pathway analysis showed that signaling pathways such as cell matrix adhesion,leukocyte transendothelial transfer,and ECM receptor interactions were significantly enriched.Conclusion:NGS screened differentially expressed genes FN1,CD44,and VCAM-1 in valve tissues of RHD patients and non-RHD patients,which may play important roles in signaling pathways such as ECM receptor interaction and leukocyte transendothelial transfer.