Effect of Curculigoside on Kainic Acid Induced Microglia Injury by Regulating JAK2/STAT3 Signaling Pathway In Vitro

The aim of this study was to investigate the effect of CCG (curculigoside) on microglia injury induced by KA (kainic acid) in vitro through regulating the JAK2 (Janus kinase-2)/STAT3 (signal transducer and activator of transcription 3) signaling pathway. HMC3 cells were randomly divided into HMC3 group, KA group, L-CCG group, M-CCG group, H-CCG group, and H-CCG+colivelin (JAK2/STAT3 pathway activator) group. The cells in the HMC3 group were not intervened, the cells in the KA group were stimulated with 600 μmol/L KA, the cells in the L-CCG, M-CCG, and H-CCG groups were added with 5, 10, and 20 μmol/L CCG, respectively, and the cells in the H-CCG+colivelin group were added with 0.5 μmol/L colivelin on the basis of the H-CCG group. The cell proliferation was detected by EdU method. Apoptosis was detected by Hochest 33258/PI method. The expression of ROS (reactive oxygen species) was detected by DCFH-DA method. DNA damage was detected by γ-H2AX (γ histone family 2A variant) immunofluorescence method. The levels of NADP+ /NADPH (nicotinamide adenine dinucleoside phosphate/reduced nicotinamide adenine dinucleotide phosphate), IL-1β (interleukin-1β), TNF-α (tumor necrosis factor-α), and IL-6 in cells were detected by kit. The expression of p-JAK2 (phosphorylated JAK2), p-STAT3 (phosphorylated STAT3), NLRP3 (NOD-like receptor thermal protein domain associated protein 3), Caspase-1 (cysteinyl aspartate specific proteinase 1), and ASC (apoptosis-associated speck-like protein) were detected by Western blot. The results showed that compared with the HMC3 group, the proliferation rate of HMC3 cells in the KA group was decreased, while the apoptosis rate, number of γ-H2AX nuclear foci, and the levels of NADP+ /NADPH, ROS, IL-1β, TNF-α, and IL-6 were increased (P<0.05). Compared with the KA group, the proliferation rate of the LCCG group, M-CCG group, and H-CCG group were increased successively, while the apoptosis rate, number of γ-H2AX nuclear foci, and the levels of NADP+ /NADPH, ROS, IL-1β, TNF-α, and IL-6 were decreased successively (P<0.05). Compared with the H-CCG group, the proliferation rate of the H-CCG+colivelin group was decreased, apoptosis rate, number of γ-H2AX nuclear foci, and the levels of NADP+ /NADPH, ROS, IL-1β, TNF-α, and IL-6 were increased successively (P<0.05). Compared with the HMC3 group, the expression of p-JAK2, p-STAT3, NLRP3, Caspase-1, and ASC in HMC3 cells in the KA group were increased (P<0.05). Compared with the KA group, the expression of p-JAK2, p-STAT3, NLRP3, Caspase-1, and ASC in the L-CCG group, M-CCG group, and H-CCG group were decreased successively (P<0.05). Compared with the H-CCG group, the expression of p-JAK2, p-STAT3, NLRP3, Caspase-1, and ASC in the H-CCG+colivelin group were increased (P<0.05). In conclusion, CCG may have a protective effect on KA-induced microglia dysfunction by inhibiting the JAK2/STAT3 pathway

CSTR: 32200.14.cjcb.2025.02.0005