Methyl tert-butyl ether (MTBE),as a widely used gasoline additive,is suspected of being environmentally toxic.MTBE accumulates mainly in adipose tissue,but its effect on obesity or obesity-related metabolic disorders has not been well understood yet.Therefore,we examined the effect of MTBE on the adipose function and the related metabolic processes with both 3T3-L1 cell line and C57BL/6J mice model.We found that exposure to MTBE at the environmental relevant concentration (100 μmol/L) could significantly induce differentiation of preadipocyte and disturb insulin-stimulated glucose uptake of mature adipocyte.The in vivo observation in male mice showed a positive correlation of visceral white adipose tissue (vWAT) expansion and cell size increase with MTBE treatment in 14 weeks.Glucose tolerance and insulin sensitivity tests demonstrated that MTBE at 1000 μg/(kg·day) disturbed the systemic glucose metabolism in a gender-specific manner,which might be partly attributed to the alterations of gut microbiota community at genus level with respect to Akkermansia,Clostridium XIVb,and Megamonas.In summary,our study characterized the effect of MTBE on adipose tissue function and glucose homeostasis in vitro and in vivo,and revealed that systemic disorders of the glucose metabolism might be modulated by the related gut microbiota.