p38MAPK在体外培养脊髓星形胶质细胞P物质刺激活化中的作用.docVIP

p38MAPK在体外培养脊髓星形胶质细胞活化中王 强，宋 波（第三军医大学西南医院全军泌尿外科研究所，重庆400038） 摘要目的 观察体外培养脊髓星形胶质细胞P物质刺激活化过程中p38MAPK活性、TNF-(、NONOS活性的变化，探讨p38MAPK活性变化与脊髓星形胶质细胞活化的关系。 方法 分离培养SPF大鼠脊髓星形胶质细胞设正常对照组（正常组）、SP刺激组（SP组10-7mol/L）、SB203580阻断p38MAPK组（SB组10μmol/L）和SP刺激+SB203580阻断p38MAPK组（SP+SB组）。WB法、RT-PCR法、ELISA法检测1,3,12,24h和36hp38MAPK、p-p38（磷酸化p38MAPK）含量及TNF-(、NO水平和NOS活性的变化。 结果脊髓星形胶质细胞GFAP 阳性表达率大于95%SP组脊髓星形胶质细胞总p38MAPK无显著变化，1h后p-p38开始升高，3h后GFAP mRNA水平显著增高，同时NO、NOS和TNF-a水平显著增高用SB203580阻断p38MAPK通路后，SP+SB组较SP组p-p38、GFAP mRNANO、NOSTNF-a水平显著降低。SB组总p38MAPKp-p38、GFAP mRNANO、NOSTNF-a水平结论 p38MAPK信号通路参与了脊髓星形胶质细胞P物质刺激活化过程，阻断p38MAPK信号通路可有效降低炎性因子水平。 关键词脊髓; 星形胶质细胞; 活化; p38MAPK 中图分类号 文献标识码A Role of p38MAPK in the substance P induced activation of spinal cord astrocytes in vitro Wangqiang，Song Bo （Center for Urology of PLA，Southwest Hospital，Third Military Medical University， Chongqing 400038， China ） Abstract Objective The changes of p38MAPK, TNF-(, NO and NOS in the activation of spinal cord astrocytes in vitro were observed in order to reveal the relationship between p38MAPK and astrocytes activation. Methods Aastrocytes, cultured from spinal cord of SPF rat and identified by immunofluorescence method with GFAP antibody, were grouped into the control group, the SP stimulus group（SP group） and the SB203580 interrupt p38MAPK group （ interrupt group）in which SP (10-7mol/L) and SB203580 (10μmol/L) were added to the supernatant for 12h. The WB method, RT-PCR method and ELISA method were used to determine the changes of p38MAPK, phosphorylated p38MAPK, GFAP mRNA, TNF-(, NO and NOS of astrocytes or supernatant at 1h,3h,12h,24h and 36h respectively. Results The GFAP positive rate of the cultured cells was higher than 95%. p38MAPK level in the SP group did not have any change, whereas p-p38 level increased significantly after 1h and GFAP mRNA, NO, NOS and TNF-a level increased after 3h. When p38MAPK pathway was inhibited by SB203580 in the SP+SB group, GFAP mRNA, NO, NOS and TNF-a was sig