中国免疫学杂志2024，Vol.40Issue(4)：698-704,713,8.DOI:10.3969/j.issn.1000-484X.2024.04.005
基于小鼠脾脏转录组测序探索脓毒症预后关键基因
Exploring key genes for prognosis of spesis based on transcriptome sequencing of mouse spleen
摘要
Abstract
Objective:To screen key differentially expressed genes(DEGs)in dead mice with sepsis by spleen high-through-put sequencing combined with bioinformatics.Methods:①A mouse sepsis model was set up by intraperitoneal injection of lipopolysac-charide(LPS),a 7-day survival curve of mice was drawn,and the modeling doses of survival group and death group were screened out.②Expressions of TNF-α,IL-1β,IL-6 and IL-10 in peripheral blood of mice in control group,survival group and death group were verified by ELISA.③High-throughput sequencing was conducted on spleens of survival group and death group,and the key genes were screened by bioinformatics analysis of DEGs.④Expressions of key genes and proteins were detected by RT-PCR and Western blot.Results:①LPS dosage in survival group was 15 mg/kg(with a mortality of 30%),and LPS dosage in death group was 30 mg/kg(with a mortality of 80%).②Expression levels of IL-6,TNF-α and IL-1β in sepsis mice were significantly higher than those of control group,while expression level of IL-10 was decreased(P<0.05).Comparison of sepsis model groups showed that levels of pro-inflammatory factors in death group were higher than those in survival group,while level of IL-10 was lower than that in survival group(P<0.05).③A total of 2999 DEGs in survival group and death group were screened out by bioinformatics,among which 1185 genes were up-regulated and 1814 genes were down-regulated.Top 5 DEGs enrichment pathways were screened out:"hematopoietic cell lineage""primary immunodeficiency""African trypanosomiasis""leishmaniasis"and"B-cell receptor signaling pathway".Ifit1,Ifit3 and Mx1 were three key genes that were screened out.④Compared with survival group,expressions of genes and proteins of Ifit1,Ifit3 and Mx1 were down-regulated in spleen tissues of the death group(P<0.05).Conclusion:By high-throughput sequencing and bioinformatics,Ifit1,Ifit3 and Mx1 are screened out as key genes related to the death outcome of sepsis,which probably influence the outcome of sepsis through the immune mechanism related to virus infection.关键词
脓毒症/脾脏/转录组学/预后/关键基因Key words
Sepsis/Spleen/Transcriptomics/Prognosis/Key genes分类
医药卫生引用本文复制引用
罗福龙,张雨婷,余亚义,胡迎春,陈睦虎,钟武..基于小鼠脾脏转录组测序探索脓毒症预后关键基因[J].中国免疫学杂志,2024,40(4):698-704,713,8.基金项目
四川省科技厅项目(2019JDPT0003,2020YFS0517) （2019JDPT0003,2020YFS0517）
泸州市-西南医科大学联合课题(2019LZXNYDJ32). （2019LZXNYDJ32）
