Objective: The objective of present study was to examine the changes of melatonin receptor agonist Neu-P11 on expression of p-IRS-1/PI3K/p-GSK3β in 3T3-L1 adipocytes and to research the variation mechanisms, observe Neu-P11 improving insulin resistance because of p-IRS-1/PI3K/p-GSK3β abnormal expression of signaling pathways leading. Thereby, we can deeperly study the impact of Neu-p11 on theinsulin signaling pathway in adipocytes from insulin resistant mice.Methods: 3T3-L1 pre-adipocytes were cultivated and differentiated in adipogenic cocktailby IBMX, DEX and insulin; Confirm the establishment of insulin resistance model. The changes of p-IRS-1/PI3K/p-GSK3βprotein expressions before and after drug action were detected by Western blot; Results: Expression of Pp-IRS-1/PI3K/p-GSK3βreduced significantly by comparison with IS (P < 0.05). Melatonin and Neu-P11 can increase obviously the expression of p-IRS-1/PI3K/p-GSK3β(P < 0.05). But Luzindole can blockthe effect of melatonin, Neu-P11 increasing p-IRS-1/PI3K/p-GSK3β (P < 0.05).Conclusions: Melatonin and Neu-P11 can increase obviouslysignificant protein the expression of insulin receptor signaling pathway(P < 0.05). Neu-P11 and Mel improve important protein expression of insulin signaling pathway by coupling with melatonin receptor 2.