资源描述：
《p38MAPK在体外培养脊髓星形胶质细胞P物质刺激活化中p38.doc》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、p38MAPK在体外培养脊髓星形胶质细胞P物质刺激活化中p38MAPK变化规律及意义的作用王强，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水平及活性、TNF-a、NO分泌水平及和NOS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大鼠乳鼠脊髓星形胶质细胞，GFAP细胞免疫荧光化学法鉴定。设正常对照组（正常组）、SP刺激组（SP组）、单纯SB203580阻断p38MAPK组（SB组）和SP刺激+SB203580阻断p38M
2、APK组（SP+SB组），SP和SB203580作用终浓度均为10-7mol/L，作用时间为12h。用WB法、RT-PCR法、ELISA法检测各组脊髓星形胶质细胞1h,3h,12h,24h和36h总p38MAPK及p-p38（磷酸化活化的p38MAPK）蛋白含量变化及TNF-a、NO分泌水平和NOS活性的变化。结果SP组脊髓星形胶质细胞总p38MAPK水平无显著变化，1h后p-p38开始显著升高，3h后GFAPmRNA水平开始显著增高，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组同时间点p-p
3、38水平、GFAPmRNA、水平，以及NO、NOS、和TNF-a水平显著降低。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该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】ARoleChangeandsignificanceofp38MAPKinthesubstancePinducedactivationofspinalcordastrocytesinvitroculturedfromspinalcordWangqiang，SongBo（Centerf
4、orUrologyofPLA，SouthwestHospital，ThirdMilitaryMedicalUniversity，Chongqing400038，China）【Abstract】ObjectiveThechangesofp38MAPK,TNF-a,NOandNOSinthesubstancePinducedactivationofspinalcordastrocytesinvitroculturedfromspinalcordwereobservedtoinvestigatetherelationshipbetweenp38MAPKanda
5、strocytesactivation.MethodsAastrocytes,culturedfromspinalcordofSPFratandidentifiedbyimmunofluorescencemethodwithGFAPantibody,weregroupedintothecontrolgroup,theSPstimulusgroup（SPgroup）andtheSB203580interruptp38MAPKgroup（interruptgroup）inwhichSP(10-7mol/L)andSB20358010(10-7mol/L)we
6、readdedtothesupernatantfor12h.TheWBmethod,RT-PCRmethodandELISAmethodwereusedtodeterminethechangesofp38MAPK,phosphorylatedp38MAPK,TNF-a,NOandNOSofastrocytesorsupernatantat1h,3h,12h,24hand36hrespectively.ResultsThecontentofp38MAPKintheSPgroupdidnothaveanychange,whereasp-p38increase
7、dsignificantlyafter1handGFAPmRNA,NO,NOSandTNF-alevelincreasedafter3h.Whenp38MAPKpathwaywasinhibitedbySB203580intheSP+SBgroup,GFAPmRNA,NO,NOSandTNF-awassignificantlyreducedcomparedwiththoseintheSPgroup.ConclusionTheseresultsindicatethatp38MAPKsignalpathwaycontributestoplaysacentra
8、lroleinPsubstance-inducedactivatonofspin
