Microglia-related retinal inflammation is associated with a variety of retinal diseases. Lycium barbarum Glycopeptide (LBGP) is the main active component of Lycium barbarum polysaccharide (LBP) isolated from Lycium barbarum, which possesses antioxidation, neuroprotection, and immunoregulation. However, whether LBGP has beneficial effects on LPS/IFN-γ (L/I) induced retinal microglial activation and its underlying mechanism remains to be elucidated. In this study, we observed the anti-inflammatory properties of LBGP in microglia activated by LPS and IFN-γ (L/I), examining both primary retinal microglia and BV2 microglial cells. Cells were pretreated with LBGP for 2h before stimulated with LPS/IFN-γ (L/I) for 24h. The dosages of LBGP for cells were evaluated by using the CCK-8 assay. The morphological changes were observed using an optical microscope. The changes in expression of cytokines and inflammatory mediators were measured with Griess assay, ELISA, qPCR, and immunofluorescence. Western blotting was used to assess MAPK, NF-κB signal transducer. LBGP pre-treatment significantly reversed L/I-induced microglia morphological alterations in area, perimeter, Feret's diameter, and roundness. LBGP significantly alleviated the L/I-induced microglial activation in primary and BV2 microglial cells. LBGP switched the M1 pro-inflammatory phenotype to the M2 anti-inflammatory phenotype by downregulating the expression of M1 markers (iNOS, IL-1β, IL-6, COX2, CD16, and CD86) and upregulating the expression of M2 markers (arginase 1 and CD206). Furthermore, LBGP attenuated the activation of NF-κB and MAPK pathways in L/I-activated BV2 cells. In summary, it is suggested that LBGP pretreatment significantly alleviated L/I-induced retinal inflammation by regulating microglial(M1/M2) polarization via NF-κB and MAPK signaling pathways.