Objective：To observed the effect of electroacupuncture on the expression of key factors as Toll like receptor 4(TLR4)，Nuclear factor κβ(NF-κβ)，Interleukin 1 receptor related kinase1(IRAK1)，Tumor necrosis factor receptor related factor 6(TRAF6)， MicroRNA146a(miR146a)in TLR4/NF-κβ signal pathway in spleen of rats with sleep deprivation.Methods：Randomly divided 32 male Wistar ratsof SPF grade into the control group， model group， diazepam group and electroacupuncture group，8 rats in each group.The control group did not receive any treatment.After the other three groups received sleep deprivation by injection of para-chlorophenylalanine(PCPA)，the diazepam group received intraperitoneal injection of diazepam and the electroacupuncture group received electroacupuncture at Bai Hui and Shen Ting.Each group was given treatment once a day.After continuous treatment of five days，detected the expression of TLR4，NF-κβ，IRAK1，TRAF6 and miR146a in spleen of rats by RT-PCR.Results：Compared with the control group，the mRNA expression of TLR4，NF-κβ，IRAK1 and TRAF6 in spleen of rats was obviously increased and the expression of miR146a in spleen of rats was increased in the model group，differences being significant(P ＜ 0.05).Compared with the model group，the mRNA expression of TLR4，NF-κβ，IRAK1 and TRAF6 in spleen of rats in the diazepam group and the electroacupuncture group was obviously decreased，the expression of miR146a in spleen of rats in the diazepam group was obviously decreased，and the expression of miR146a in spleen of rats in the electroacupuncture group was obviously increased，differences being significant(P＜0.05).Conclusion：The therapy of electroacupuncture at Bai Hui and Shen Ting can improve the expression of miR146a in spleen of rats with sleep deprivation so as to negatively regulate TLR4 signal pathway，and eventually regulate innate immune response and relive the injury of body by sleep deprivation.