Screening and validation of key genes for hypoxia-induced renal inflammatory reaction in mice by transcriptome sequencing and bioinformatics
10.12007/j.issn.0258-4646.2024.12.003
- VernacularTitle:基于转录组学测序与生物信息学筛选低氧诱导小鼠肾脏炎症反应的关键基因并验证
- Author:
Xintong XU
1
;
Qifu LONG
1
;
Ying HU
1
;
Ruhan JIA
1
;
Ruxue MA
1
;
Sheng YONG
1
Author Information
1. 青海大学医学院基础医学部,西宁 810016
- Publication Type:Journal Article
- Keywords:
plateau hypoxia;
kidney;
inflammatory reaction;
transcriptome sequencing;
bioinformatics
- From:
Journal of China Medical University
2024;53(12):1071-1079
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen and validate the key genes involved in hypoxia-induced inflammatory reactions in mice using transcrip-tome sequencing and bioinformatics. Methods C57/BL6 mice were bred at altitudes of 4200 m and 400 m,and mouse models were con-structed for the plateau hypoxia (HKT) group and the plain normoxia (PKC) group. Kidney tissues were aseptically removed after 30 days,renal pathological changes were analyzed by HE staining,blood gas analysis and renal index changes were measured in the mice under hypoxia. The kidney tissues of mice in the HKT and PKC groups were analyzed using transcriptome sequencing,key genes were screened using bioinformatics technology,and these genes were verified using real-time quantitative reverse transcription PCR (RT-qPCR) and Western blotting. Results HE staining showed glomerular atrophy in mice in the HKT group compared with the PKC group,and a decrease in blood gas analysis and renal index occurred in mice exposed to hypoxia. Transcriptome sequencing analysis revealed 3007 differentially expressed genes (DEGs) in the HKT group,of which 123 were inflammation-related DEGs (IR-DEGs). GO and KEGG enrichment analyses of IR-DEGs showed significant enrichment in inflammation-related signaling pathways,such as cytokine-cytokine receptor interactions and chemokines. The results of the protein-protein interaction (PPI) network construction of IR-DEGs showed that six hub genes,STAT3,TLR7,CD68,NFKBIA,LEP,and APOE,were identified,and the mRNA expression of these six genes was upregulated according to RT-qPCR results,which was in agreement with the results of transcriptome sequencing. Western blotting showed that CD68,NFKBIA,LEP,TLR7,and APOE expression was upregulated while STAT3 expression was downregulated. Conclusion STAT3,CD68,NFKBIA,LEP,TLR7,and APOE are the key genes involved in hypoxia-induced inflammatory reactions. A hypoxic environment induced inflammatory reactions in mouse kidney tissues by upregulating the expression of TLR7,CD68,STAT3,LEP,and APOE.
