Objective Insulin resistance is closely related to chronic low-grade inflammation in adipose tissue, liver and muscle with following activation of macrophage proinflammatory pathways and cytokine secretion. At the molecular level, insulin resistance is promoted by a transition in macrophage polarization from an M2 (alternatively activated macrophages) state controlled by STAT6 and PPARs to a M1 (classically activatedmacrophages) state driven by NF-κB, and other signal transcription factors. However, key factors of function and polarization in macrophage in metabolism are unclear. Methods To determine the role of mitochondria of macrophages in IR, we have generated macrophage specific OxPhos complex-deficient (LysM^cre CRIF1^flox/flox) mice, because CRIF1 plays a critical role in the integration of OxPhos polypeptides into the mitochondrial membrane in mammals. CRIF1^flox/flox strain was intercrossed with LysM^cre strain to generate LysM^cre CRIF1^flox/flox. Macrophages were stimulated with cytokines or LPS for 30 minutes or 18 hours, and western blot analysis, Q-PCR and ELISA were performed. For animal study, body weight, GTT and ITT of mice were monitored after 10 weeks on HFD. Results The proinflammatory cytokines, reactive oxygen species (ROS) were significantly increased in bone marrow-derived macrophages (BMDMs) from LysM^cre CRIF1^flox/flox mice compared with control. Moreover, LPS and Interferon (IFN)-γ stimulation of BMDM from LysM^cre CRIF1^flox/flox mice caused a marked potentiation of TNFα, IL-6 and iNOS response, it is direct evidence that OXPHOS complex dysfunction promotes and amplifies M1-like phenotype. However, M2-like activation was impaired in IL-4 stimulated BMDM from LysM^cre CRIF1^flox/flox mice. In addition to, insulin-stimulated phosphorylation of AKT was markedly decreased in BMDM from LysM^cre CRIF1^flox/flox mice. In animal study, Body weights were similar in both groups, but LysM^cre CRIF1^flox/flox mice were significantly more glucose intolerant after a 10 week HFD challenge. Conclusion Taken together, these date demonstrate that OxPhos complex has a critical role in macrophage M1 and M2-like polarization and regulates metabolic homeostasis.