〔Abstract〕 Objective Based on metabolomics and transcriptomics analysis, to explore the molecular mechanism of spleen inflammation induced by high altitude hypoxia in mice through NOD-like receptor signaling pathway. Methods C57BL/6 mice were raised at an altitude of 400 m and 4 200 m respectively, with five mice in each group, and spleen tissues were collected after 30 days. Differential metabolites and differentially expressed genes in key pathways were screened by metabolomics and transcriptome analysis and correlation KEGG enrichment analysis, and the related network interaction diagram of differential metabolites and differentially expressed genes in key pathways was constructed and verified by RT-qPCR. Results Metabolomics analysis showed that 133 differential metabolites were screened from in the plain spleen control group(PSC group) and the plateau spleen test group(HST group), 95 of which were up-regulated while 38 of which were down-regulated. KEGG enrichment analysis showed that they were mainly involved in NOD-like receptor signaling pathway, HIF-1 signaling pathway, cholesterol metabolism and other metabolic pathways. The results of transcriptome analysis showed that a total of 4213 differentially expressed genes were identified in PSC group and HST group, including 1947 up-regulated genes and 2266 down-regulated genes. KEGG was enriched in 173 signaling pathways, including NOD-like receptor signaling pathway, MAPK signaling pathway, NF-κB signaling pathway and other pathways. Comprehensive analysis showed that the differential metabolites and differentially expressed genes were obviously enriched in NOD-like receptor signaling pathway. Therefore, the correlation network interaction map was constructed for the differential metabolites ATP and differentially expressed genes in NOD-like receptor signaling pathway. RT-qPCR results showed that compared with PSC group, the expression levels of DEGs related to NOD1 and NOD2(CHUK, TAB3, MAPK8) in the signaling pathway of NOD-like receptor and NLRP1-CASP1 pathway(NLRP1b, CASP1) in HST group were significantly enhanced. The mRNA expression levels of downstream inflammatory factors IL-6, IL-1 β, IL-18, INF-γ and TNF-α were up-regulated and differentially expressed. Conclusion Based on the combined analysis of metabolomics and transcriptomics, it was found that hypoxia stimulation at high altitude may affect the NOD-like receptor signaling pathway in vivo, and the differential metabolite ATP is positively correlated with the differential key genes in the pathway. ATP mediates the release of downstream inflammatory factors by activating NOD1, NOD2 pathways and NLRP1 inflammable-CASP1 pathways. Inflammatory response occurred in spleen tissue of mice.