[Abstract] Objective: To investigate the role of Toll-like receptors (TLRs) / NF-κB signaling pathway in methamphetamine (METH) -induced primary microglial activation and inflammatory reaction. Methods: Primary microglial cells were isolated and cultured, then the purity of primary microglia was detected by immunofluorescence. Cells were treated with METH (300 μM) for0 min, 15 min, 30 min, 1 h, 3 h, 6 h, 12 h and 24 h, and the activation of primary microglia and NF-κB pathway was subjected to western blot. The inflammatory factors (IL-6, TNF-α, IL-1β) and TLR1-9,11 mRNA levels were detected byReal-time PCR. Results: The purity of primary microglial cells was above 95%. After exposure to METH, the primary microglia activation marker IBA-1was increasedand the NF-κB pathway was activated. In addition, the mRNA levels of inflammatory factors (IL-6, TNF-α, IL-1β) and TLR2、4-5、7-9、11 were significantly increased (P <0.05),while the level of TLR1 mRNA was significantly reduced. Conclusion: METH can activate primary microglial cells and promote the release of inflammatory factors by regulating the TLRs/NF-κB signaling pathway. Therefore, TLRs can be a potential target for METH neuritis response interventionwith certain therapeutic significance.