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The mechanism of fat soluble bioactives on activation of AMPK, normalization of insulin and serum proteome on obesity is not yet clear. This study investigated the effect of wheat bran oil concentrate on adipokines, pro-inflammatory markers and transcription factors in relation to differentially expressed serum proteins using two-dimensional electrophoresis (2DE) approach-in high fat diet (HFC) fed C57BL6 mice.The HFC group exhibited a significant increase in body weight, insulin level and lipid profile. The ELISA and Western blot analysis of serum proteins of HFC group also exhibited increased level of inflammatory markers (TNF-α, TGF-β and interleukin-1β), tissue transcription factors (SREBP-2 & PPARγ) and were validated by gene expression and immunohistochemistry. However, the treatment with WBA 1 and 2% significantly lessened lipid droplets in 3T3-L1 cells. Reduced insulin level, down-regulated transcription factors and associated inflammatory cytokines by activating AMPK in HFC-fed mice. Furthermore, WBA treatment directly reversed differentially expressed recombinant proteins of KCTD family and Zinc finger protein 236 isoform.Bioactive fraction of WBA had beneficial effect on ameliorating obesity related complications mainly through phosphorylation of AMPK and inflammatory cytokines which was authenticated by proteomic analysis.