Abstract: 【Aim】 To analyze the contents of heavy metals and metabolomics of Eucriotettix oculatus living in metal and non-metal mining areas, and to explore the effects of combined pollution of heavy metals on the accumulation and metabolomics of heavy metals in E. oculatus. 【Methods】 ICP-MS method was used to determine the content of heavy metals in E. oculatus adults. At the same time, based on UPLC-MS/MS detection platform, selfbuilt database and multivariate statistical analysis, the differences of metabolites in the intestine of E. oculatus adults between the metal mining area and the non-metal mining area were analyzed, and KEGG database was used to conduct annotation and pathway enrichment analysis of the differential metabolites. 【Results】 The contents of nine heavy metals in E. oculatus adults in the metal mining area were 0.4-212.4 times as high as those in the non-metal mining area. The results of multivariate statistical analysis showed that the contents of 112 metabolites changed significantly in the intestine of E. oculatus adults in the metal mining area, mainly including amino acids, fatty acyls, organic acids, nucleotides and benzenes. KEGG annotation and pathway enrichment analysis showed that there were 49 significantly differential metabolites that could be annotated, and 40 significantly differential metabolites were related to metabolic pathways. The most significant enrichment pathways included thyroid hormone synthesis pathway, oxytocin signaling pathway, bile secretion pathway and tyrosine metabolism pathway. 【Conclusion】 There are multiple kinds of heavy metals accumulated in E. oculatus adults living in the environment polluted by multiple heavy metals. Heavy metals can change the composition of metabolites in the intestine of E. oculatus, and the change of some metabolites may be a strategy for E. oculatus to adapt to the habitat polluted by multiple heavy metals.

Key words: Eucriotettix oculatus, heavy metals, combined pollution, intestinal tissue, metabolomics, metabolic pathway