Cumulative evidence has suggested that many antipsychotics cause metabolic abnormalities. Adipose tissue insulin resistance (Adipo-IR) contributes to the development and progress of metabolic abnormalities including fatty liver by inducing excessive free fatty acid release from adipose tissue. Sulpiride is an old antipsychotic still frequently used in many developing countries. However, its adverse metabolic effects remain poorly understood. Here, chronic administration of sulpiride (80 mg/kg, subcutaneously, once daily for 6 weeks) elevated fasting insulin concentration and the index of the homeostasis model assessment of insulin resistance in rats. More importantly, sulpiride increased hepatic triglyceride accumulation and Oil Red O-stained area, indicating the induction of fatty liver by sulpiride. Sulpiride also increased plasma non-esterified fatty acid concentrations at the baseline and during an oral glucose tolerance test, the Adipo-IR index, and adipocyte size. Adipose gene expression profile revealed that sulpiride decreased mRNA and protein expression of insulin receptor substrate (IRS)-1, but not IRS-2. Furthermore, sulpiride increased phosphorylation of both Ser307 in IRS-1 and Ser473 in Akt at baseline. Co-treatment with bromocriptine (a dopamine D2 receptor agonist) attenuated sulpiride-induced hyperprolactinemia, but it was without effect on insulin resistance and fatty liver. Therefore, the present results suggest that sulpiride induces fatty liver in rats via phosphorylation of IRS-1 at Ser307-mediated adipose tissue insulin resistance, in which dopamine D2 receptor is possibly not involved. Our findings may provide new insights into the mechanisms underlying the steatotic effect of the old antipsychotic. Copyright © 2018. Published by Elsevier Inc.