高原低氧环境抑制小鼠脾脏组织碳代谢的分子机制  

Molecular Mechanism of Carbon Metabolism Inhibition in Spleen Tissues of Mice Under High Altitude Hypoxia Environment

作　　者：陈晓晨 胡英 许玉珍 龙启福 马茹雪 永胜 CHEN Xiaochen;HU Ying;XU Yuzhen;LONG Qifu;MA Ruxue;YONG Sheng(Department of Immunology,School of Basic Medicine,Qinghai University School of Medical,Xining 810016,China)

机构地区：[1]青海大学医学院基础医学部免疫学教研室,青海西宁810016

出　　处：《中山大学学报（医学科学版）》2024年第5期806-817,共12页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(82060295);青海省科技计划项目基金(2023-ZJ-771)。

摘　　要：【目的】探究低氧暴露对小鼠脾脏组织碳代谢通路的调控机制。【方法】将C57BL/6小鼠分别在海拔400 m和4200 m饲养,每组5只,30 d后无菌取出脾脏组织,通过转录组测序,蛋白质组学和非靶向代谢组学分析差异表达基因、差异蛋白以及差异代谢物,利用GO和KEGG富集分析,挖掘关键通路,并通过RT-qPCR和Western blot对通路中关键基因和蛋白进行验证。【结果】转录组测序发现,低氧暴露下有4213个基因出现显著性差异表达,其中1947个基因表达上调,2266个基因表达下调;差异表达蛋白分析发现166个蛋白表达上调,39个蛋白表达下调;非靶向代谢组学结果表明,高原低氧条件下存在133个差异代谢物,95个上调,38个下调;通过KEGG富集分析表明,差异表达基因、差异蛋白以及差异代谢物共同富集到碳代谢通路,因此,对碳代谢通路中的关键基因和蛋白进行验证,发现碳代谢通路中PGAM2、ENO3、PRPS2、PGLS、RPE、IDH3A、SUCLA2、MDH2的mRNA和蛋白表达量显著下调。【结论】高原低氧环境通过抑制机体碳代谢通路,减弱了糖酵解、三羧酸循环、磷酸戊糖途径,致使机体发生氧化应激、能量代谢失衡等。【Objective】To explore the regulatory mechanism of hypoxia exposure on carbon metabolism pathway in spleen of mice.【Methods】C57BL/6 mice were raised at altitudes of 400 m and 4200 m,with 5 mice in each group.After 30 days,spleen tissues were aseptically removed for analysis of differentially expressed genes,proteins,and metabolites using transcriptome sequencing,proteomics,and non-targeted metabolomics.GO and KEGG enrichment analysis were conducted to explore key pathways.The key genes and protein in the pathway were validated by RT-qPCR and Western blot.【Results】Transcriptome sequencing revealed a significant difference in the expression of 4213 genes in hypoxic exposure,of which 1947 were up-regulated and 2266 were down-regulated.The analysis of differentially expressed proteins showed that 166 proteins were up-regulated and 39 proteins were down-regulated.The results of non-targeted metabolomics showed that 133 different metabolites were screened under high altitude hypoxia condition,of which 95 were up-regulated and 38 were down-regulated.KEGG enrichment analysis showed that differentially expressed genes,differentially expressed proteins and differentially expressed metabolites were enriched into the carbon metabolic pathway.Therefore,the key genes and proteins in the carbon metabolic pathway were verified.The mRNA and protein expressions of PGAM 2、ENO 3、PRPS 2、PGLS、RPE、IDH 3 A、SUCLA 2 and MDH 2 were significantly down-regulated in the carbon metabolism pathway.【Conclusion】Low oxygen environment at high altitude weakens glycolysis,tricarboxylic acid cycle and pentose phosphate pathway by inhibiting the carbon metabolism pathway of the body,resulting in oxidative stress and energy metabolism imbalance.

关 键 词：多组学 低氧 脾脏 碳代谢 能量 小鼠 

分 类 号：R34[医药卫生—基础医学]