补阳还五汤通过抑制JAK2/STAT3通路促进BV2小胶质细胞M2极化以减轻炎症反应OA

Objective:To investigate the effect of Buyang H uanwu decoction(BYHWD)on the polarization of BV2 microglial cells induced by lipopolysaccharide(LPS)by reg-ulating the JAK2/STAT3 signaling pathway.Methods:A neuroinflammatory injury model was established in BV2 microglial cells using LPS.The cells were divided into blank group,model group,BYHWD group,Colivelin group(STAT3 activator),and BYHWD+Colivelin group.Cell viability was assessed using the Cell Counting Kit-8(CCK-8)assay.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the secretion levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-10(IL-10),and in-terleukin-4(IL-4).Immunofluorescence co-expression analysis of F4/80/iNOS and F4/80/CD206 was used to observe BV2 cell activation.Western Blot was performed to detect the expression of iNOS,IL-1β,Arg1,Fizz1,and proteins related to the JAK2/STAT3 signaling pathway.Results:Compared with the blank group,the model group exhibited significantly decreased cell viability,levels of the anti-inflammatory cytokines IL-10 and IL-4,the proportion of F4/80 and CD206 doub-le-positive cells,and the protein expression levels of Fizz1 and Arg1(P＜0.05).Conversely,the levels of the pro-inflam-matory cytokines IL-6 and TNF-α,the proportion of F4/80 and iNOS double-positive cells,and the protein expression lev-els of iNOS,IL-1β,p-JAK2/JAK2,and p-STAT3/STAT3 were significantly increased(P＜0.05).Compared with the model group,the BYHWD group showed significantly increased cell viability,levels of IL-10 and IL-4,the proportion of F4/80 and CD206 double-positive cells,and the protein expression levels of Fizz1 and Arg1(P＜0.05),while the levels ofIL-6 and TNF-α,the proportion of F4/80 and iNOS double-positive cells,and the protein expression levels of iNOS,IL-1β,p-JAK2/JAK2,and p-STAT3/STAT3 were significantly decreased(P＜0.05).Compared with the BYHWD group,the Colivelin group exhibited significantly decreased cell viability,levels of IL-10 and IL-4,the proportion of F4/80 and CD206 double-positive cells,and the protein expression levels of Fizz1 and Arg1(P＜0.05),along with significantly increased levels of IL-6 and TNF-α,the proportion of F4/80 and iNOS double-positive cells,and the protein expression levels of iNOS,IL-iβ,p-JAK2/JAK2,and p-STAT3/STAT3(P＜0.05).Compared with the Colivelin group,the BYHWD+Colivelin group showed significantly increased cell viability,levels of IL-10 and IL-4,the proportion of F4/80 and CD206 double-positive cells,and the protein expression levels of Fizz1 and Arg1(P＜0.05),while the levels of IL-6 and TNF-α,the proportion of F4/80 and iNOS double-positive cells,and the protein expression levels of iNOS,IL-1β,p-JAK2/JAK2,and p-STAT 3/STAT 3 were significantly decreased(P＜0.05).Conclusion:BYHWD may inhibit the activation of the JAK2/STAT 3 signaling pathway,thereby reducing LPS-induced M1-type activation of BV2 microglia and promoting their polarization toward the M2 phenotype.This provides experimental evidence for the clinical treatment of neuroinflammation following spinal cord injury(SCI).