急性低氧调控肝细胞线粒体碳代谢通路关键基因的筛选与机制研究OA
Screening and mechanistic study of key genes regulating the mitochondrial carbon metabolism pathway in hepatocytes under acute hypoxia
目的:本研究旨在结合生物信息学探究急性低氧暴露下肝细胞线粒体碳代谢通路中的关键基因及其调控机制.方法:将人肝细胞系HL7702分别培养在常氧(95%的空气和5%CO2)和低氧(1%O2、94%N2和5%CO2)培养箱中,分别培养24 h后提取RNA制作表达谱基因芯片,通过微阵列实验得到的差异基因与线粒体数据库进行整合分析,分别对差异基因和差异基因与线粒体功能相关的交集基因进行GO与KEGG富集分析挖掘关键通路,并对差异基因与线粒体功能相关的交集基因构建蛋白质相互作用网络进一步筛选关键基因.用小鼠肝细胞系AML12进行低氧造模并验证筛选出的关键基因以及参与线粒体关键通路中的基因.结果:与常氧组相比,低氧组共鉴定出47 958个非显著差异基因和1 437个差异基因,其中864个基因表达上调,573个表达基因下调;对差异基因进行KEGG富集分析结果显著富集在代谢通路、氨基酸合成、碳代谢、辅酶生物合成等通路中;对差异基因与线粒体功能相关的交集基因进行KEGG富集分析发现,主要富集在碳代谢等通路中;PPI中鉴定出两个关键基因FH和HSPD1,并发现急性低氧下FH和HSPD1的mRNA和蛋白表达量显著上调;KEGG富集中共同富集到碳代谢通路中,发现急性低氧胁迫影响线粒体碳代谢通路中GCSH、AGXT、FH、GPT2、IDH2、HIBCH基因的表达.结论:急性低氧胁迫通过上调FH和HSPD1调节肝脏组织线粒体的功能,并通过影响GCSH、AGXT、FH、GPT2、IDH2、HIBCH基因的表达影响肝脏组织中线粒体碳代谢通路应对急性低氧暴露,为进一步理解急性低氧下肝细胞的相关疾病提供了新的视角.
Objective:To explore key genes and the regulatory mechanisms in the mitochondrial carbon metabolism pathway of hepatocytes under acute hypoxic exposure,using bioinformatics approaches.Methods:The human liver cell line HL7702 was cultured under normoxic conditions(95%air and 5%CO2)and hypoxic conditions(1%O2,94%N2,and 5%CO2)in separate incubators.After 24 hours of cultivation,RNA was extracted to prepare expression profile gene chips.The differential genes ob-tained from microarray experiments were integrated with a mitochondrial database for analysis.GO and KEGG enrichment analy-ses were conducted on the differential genes and the intersection genes related to mitochondrial function to excavate key pathways.A protein-protein interaction network was constructed for the differential genes and intersection genes related to mitochondrial func-tion to further screen for key genes.The mouse liver cell line AML12 was used to model hypoxia and validate the selected key genes and those involved in key mitochondrial pathways.Results:Compared to the normoxic group,the hypoxic group identified a total of 47 958 non-significant differential genes and 1 437 differential genes,among which 864 genes were upregulated and 573 genes were downregulated.KEGG enrichment analysis of the differential genes significantly enriched in metabolic pathways,ami-no acid synthesis,carbon metabolism,and coenzyme biosynthesis pathways.KEGG enrichment analysis of the intersection genes related to mitochondrial function found that they were mainly enriched in pathways such as carbon metabolism.In the protein-protein interaction(PPI)network,two key genes,FH and HSPD1,were identified,and it was found that the mRNA and protein expression levels of FH and HSPD1 were significantly upregulated under acute hypoxia.KEGG enrichment analysis,which was common to both differential genes and intersection genes related to mitochondrial function,revealed that acute hypoxia affects the expression of genes in the mitochondrial carbon metabolism pathway,including GCSH,AGXT,FH,GPT2,IDH2,and HIBCH.Conclusion:Acute hypoxia stress regulates mitochondrial function in liver tissue by upregulating FH and HSPD1 and affects the expression of GCSH,AGXT,FH,GPT2,IDH2,and HIBCH genes in the mitochondrial carbon metabolism pathway to cope with acute hypoxic exposure.These findings provide new insights for further understanding of liver cell-related dis-eases under acute hypoxia conditions.
李俊秀;台艳奇;邓钊红;徐金芳;胥瑾
青海大学医学院 基础医学部,青海 西宁 810016青海大学医学院 基础医学部,青海 西宁 810016青海大学医学院 基础医学部,青海 西宁 810016青海大学医学院 中医系,青海 西宁 810016青海大学医学院 基础医学部,青海 西宁 810016
医药卫生
急性低氧线粒体碳代谢生物信息学
Acute hypoxiaMitochondriaCarbon metabolismBioinformatics
《海南医科大学学报》 2026 (3)
179-187,9
This study was supported by the National Natural Science Foundation of China(82360333)Science and Technology Plan Project of Qinghai Province(2023-ZJ-792) 国家自然科学基金(82360333)青海省科技计划项目(2023-ZJ-792)
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