p38 MAPK-iNOS-NO通路介导化学性缺氧对PC12细胞的损伤作用  

p38 MAPK-iNOS-NO pathway mediates chemical hypoxia-induced injury in PC12 cells

作　　者：廖新学[1,2] 阮经文[3] 兰爱平[4] 王秀玉[4] 董小变[1,2] 胡芬[4] 杨战利[4] 王礼春[1,2] 陈培熹[4] 冯鉴强[4] 

机构地区：[1]中山大学附属第一医院心血管内科 [2]中山大学附属第一医院高血压血管病科 [3]中山大学附属第一医院针灸科 [4]中山大学中山医学院生理教研室,广东广州510080

出　　处：《中国病理生理杂志》2010年第12期2410-2414,共5页Chinese Journal of Pathophysiology

基　　金：广东省科技计划资助项目(No.2009B080701014;No.2007B080701030)

摘　　要：目的:探讨诱导型一氧化氮合酶/一氧化氮(iNOS/NO)是否介导了氯化钴(CoCl2)引起的PC12细胞损伤及p38 MAPK对其调节作用。方法:应用化学性低氧模拟剂CoCl2处理PC12细胞建立化学性缺氧损伤模型。应用细胞计数试剂盒-8(CCK-8)比色法检测细胞存活率;Hochest33258核染色法观察细胞凋亡的形态学改变;Western blotting法检测iNOS蛋白的表达水平;Griess试剂盒检测细胞培养基中亚硝酸盐(NO的代谢物)的浓度。结果:应用600μmol/L CoCl2处理PC12细胞24 h可使iNOS表达明显增多;应用600μmol/L CoCl2处理PC12细胞24 h和48 h可使细胞培养基里NO增多;在CoCl2损伤PC12细胞前60 min应用iNOS抑制剂L-canavanine(10μmol/L)预处理能保护PC12细胞对抗600μmol/L CoCl2引起的损伤,使细胞存活率升高,凋亡细胞数目减少;SB203580(p38 MAPK选择性抑制剂)预处理60 min可下调CoCl2引起的iNOS高表达。结论:p38 MAPK-iNOS-NO通路介导CoCl2引起PC12细胞的损伤作用。AIM：To investigate the possibility that p38 MAPK-iNOS-NO pathway mediates the chemical hypoxia-induced injury in PC12 cells.METHODS： PC12 cells were treated with cobalt chloride（CoCl2,600 μmol/L） to set up a chemical hypoxia-induced cellular injury model.Cell viability was tested by cell counter kit-8（CCK-8）.The morphological changes of the apoptotic cells were detected by Hochest33258 staining.The expression of iNOS was determined by Western blotting.Nitrite accumulation,an indicator of NO production,was measured in cell culture supernatants by Griess reagent assay.RESULTS： Exposure of PC12 cells to CoCl2 for 24 h significantly enhanced iNOS expression.Exposure of PC12 cells to CoCl2 for 24 h and 48 h significantly enhanced nitrite accumulation.Pretreatment with L-canavanine（an inhibitor of iNOS,5-20 μmol/L） for 60 min prior to exposure of PC12 cells to CoCl2 protected PC12 cells against the injuries induced by CoCl2 with enhanced cell viability and decreased amount of apoptotic cells.Pretreatment with SB203580（an inhibitor of p38 MAPK） for 60 min prior to exposure of PC12 cells to CoCl2 down-regulated the expression of iNOS induced by CoCl2.CONCLUSION： p38 MAPK-iNOS-NO pathway mediates CoCl2-induced injuries in PC12 cells.

关 键 词：氯化钴 一氧化氮合酶 一氧化氮 刀豆氨酸 化学性缺氧 

分 类 号：R33[医药卫生—人体生理学] R743[医药卫生—基础医学]