CREB和NF-κB在p38MAPK所致脊髓星形胶质细胞活化中的作用(英文)

The Role of CREB and NF-κB in the p38MAPK-induced Activation of Spinal Cord Astrocytes

目的:探讨CREB和NF-κB在p38MAPK所致脊髓星形胶质细胞活化中的作用,明确脊髓星形胶质细胞活化中p38MAPK细胞信号转导途径的作用。方法:分离培养SPF大鼠脊髓星形胶质细胞,设正常组、SP刺激组(SP组,10-7mol/L)、SP刺激+SB203580(10μmol/L)阻断p38MAPK组(SP+SB组)、SP刺激+PD98059(10μmol/L)阻断CREB组(SP+PD组)、SP刺激+SN50(10μmol/L)阻断NF-κB(SP+SN组)。WB法、免疫荧光法、ELISA法检测12 h和24 h时p-p38、p-CREB、NF-κBp65水平及GFAP、TNF-、IL-1β水平变化。结果:SP组脊髓星形胶质细胞p-p38、p-CREB、NF-κBp65显著升高,GFAP水平显著增高,同时TNF-和IL-1β水平显著增高。与SP组比较,用SB203580阻断p38MAPK通路后,SP+SB组p-p38、p-CREB、NF-κBp65显著降低,GFAP、TNF-和IL-1β水平显著降低。用PD98059阻断CREB通路后,SP+PD组p-p38、NF-κBp65无显著变化,p-CREB显著降低,GFAP水平降低,同时TNF-和IL-1β水平降低。用SN50阻断NF-κB通路后,SP+SN组p-p38、p-CREB无显著变化,NF-κBp65显著降低,GFAP水平降低,同时TNF-和IL-1β水平降低。结论:体外培养中,SP刺激后脊髓星形胶质细胞显著活化,p38MAPK活化后通过CREB及NF-κB信号途径导致胶质细胞炎性因子水平显著升高。

Objective：To investigate the role of CREB and NF-κB in the p38MAPK-induced activation of spinal cord astrocytes,p38MAPK signal transduction pathway in the activation of spinal cord astrocytes cell.Methods：Aastrocytes,cultured from spinal cord ofSPF rat,were grouped into the normal group,the SP stimulus group（SP group）,the SP stimulus＋SB203580 interrupt group（SP＋SBgroup）,the SP stimulus＋PD98059 interrupt group（SP＋PD group） and the SP stimulus＋SN50 interrupt group（SP＋SN group）in whichSP（10-7mol/L）,SB203580（10 μmol/L）,PD98059（10 μmol/L） and SN50（10 μmol/L） were added to the supernatant for 12h.TheWB method,immunofluorescence method and ELISA method were used to determine the changes of p-p38,p-CREB,NF-κBp65,GFAP,TNF-,IL-1β of astrocytes or supernatant at 12h and 24h.Results：p-p38,p-CREB,NF-κBp65 level in the SP group increased significantly.In the same time,GFAP,TNF-a and IL-1β level increased significantly too.When p38MAPK pathway was inhibited by SB203580 in the SP＋SB group,p-p38,p-CREB,NF-κBp65 level and GFAP,TNF-a and IL-1β was significantly reduced compared with those in the SP group.In the SP＋PD group,p-CREB level was significantly reduced compared with those in the SP group.In theSP＋SN group,NF-κBp65 level was significantly reduced compared with those in the SP group.Conclusion：Astrocytes from spinal cordwere significant activated after stimulated by SP in vitro,Inflammatory factors levels from glial cells were significantly increased through CREB and NF-κB signaling pathways after p38MAPK activation.