基于转录组学研究高原低氧对小鼠T、B淋巴细胞信号通路的影响

Transcriptomics-based study of the effects of high altitude hypoxia on T and B lymphocyte signaling pathways in mice

目的 基于转录组学测序技术探究高原低氧(HST)胁迫下T、B淋巴细胞信号通路中枢纽基因和关键调控通路响应的分子机制。方法 分别在海拔400 m[平原常氧组(PSC组)]和4 200 m(HST组)处饲养C57BL/6小鼠30 d后取脾脏组织,采用RNA-Seq进行转录组测序,得到差异基因进行基因本体(GO)功能注释和京都基因和基因组数据库(KEGG)通路分析,使用实时荧光定量聚合酶链反应(RT-qPCR)验证。结果 相比于PSC组,HST组中1 947个差异基因表达上调,2 266个差异基因表达下调,差异均有统计学意义(P<0.05),且KEGG富集分析发现,HST胁迫下主要通过T、B细胞受体、磷脂酰肌醇-3激酶-蛋白激酶B(PI3K-Akt)、丝裂原活化蛋白激酶(MAPK)、核因子κB(NF-κB)等信号通路调控异常引起免疫炎症的发生发展。结论 HST胁迫可能通过激活PI3K-Akt通路,抑制p38 MAPK和NF-κB通路,诱导T、B细胞受体信号通路响应HST胁迫的转录调控分子机制,引起免疫调节和炎症反应的发生。

Objective To explore the molecular mechanism of the response of key genes and key regula-tory pathways in T and B lymphocyte signaling pathways under high altitude hypoxia(HST)stress based on transcriptomic sequencing technology.Methods C57BL/6 mice were raised at 400 meters[the plain normoxia group(PSC group)]and 4200 meters(the HST group)above sea level for 30 days,respectively.RNA extrac-ted from mouse spleen tissue was used for transcriptome sequencing,and differentially expressed genes were obtained for GO functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway anal-ysis,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to verify the differen-tially expressed genes.Results Compared with the PSC group,1947 differentially expressed genes were up-regulated and 2266 differentially expressed genes were down-regulated in the HST group,the differences were statistically significant(P<0.05).KEGG enrichment analysis showed that abnormal regulation of differential-ly expressed genes between the two groups under HSP stress caused the occurrence and development of im-mune inflammation,mainly through T cell receptor,B cell receptor,PI3K-Akt,MAPK,NF-κB and other sig-naling pathways.Conclusion HSP stress may induce T and B cell receptor signaling pathways to respond to transcriptional regulatory molecular mechanisms by activating PI3K-Akt pathway,inhibiting p38 MAPK and NF-κB pathways,and causing immune regulation and inflammatory response.