p38丝裂原活化蛋白激酶在β淀粉样肽+缺氧缺糖所致SHSY5Y细胞损伤中的作用

目的探讨p38丝裂原活化蛋白激酶(p38MAPK)活性在β淀粉样肽(Aβ)+缺氧缺糖(OGD)所致SHSY5Y细胞损伤中的作用。方法体外培养SHSY5Y神经母细胞瘤株,以OGD、伴或不伴Aβ(5μmol/L)处理,以MTT检测SHSY5Y细胞活性,Westernblot方法检测p38MAPK活性,fluo-3/AM的荧光强度检测SHSY5Y细胞内钙离子的浓度。结果与对照组相比,OGD、Aβ、OGD+Aβ组SHSY5Y细胞活性显著下降,分别为对照组的(63±4)%、(68±7)%、(56±1)%,(P<0.05或P<0.01);p38MAPK活性在OGD、Aβ、OGD+Aβ组显著升高,分别为对照组的(121±3)%、(132±4)%、(165±5)%,(P<0.05或P<0.01);fluo-3/AM的荧光强度在OGD、Aβ、OGD+Aβ组显著升高,分别为照组的(169±4)%、(176±4)%、(220±7)%,(P<0.05或P<0.01)。结论与OGD、Aβ比较,OGD+Aβ对SHSY5Y细胞具有更显著的细胞毒性作用。OGD+Aβ通过激活p38MAPK,进而促进细胞内钙超载,最终导致SHSY5Y细胞死亡。

The role of p38 mitogen activated protein kinase in SHSY5Y cell injury caused by beta amyloid and oxygen glucose deprivation

Objective To investigate the role of p38 mitogen activated protein kinase （p38MAPK） in SHSY5Y cell injury caused by beta-amyloid （Aβ） and oxygen glucose deprivation （OGD）. Methods The SHSYSY neuroblastoma cells were exposed to OGD, with or without Aβ （5μmol/L）. The cell viability was detected by the MTF assay, activity of p38MAPK was detected by Western blot method, and intracellular concentration of calcium was evaluated via the fluorescence intensity of fluo 3/AM.Results Compared with the control group,the MTF value was（63±4）% ,（68±7）% ,（56± 1）% in the OGD, Aβ and OGD ＋ Aβ groups respectively, significantly lower than that that in control group （ P 〈 0.05 or P 〈 0.01 ）. The activity of p38MAPK was（ 121 ± 3） %, （ 132± 4） %, （ 165± 5） % in the OGD,Aβ and OGD ＋ Aβ groups respectively, significantly higher than that in control group（ P 〈 0.05 or P 〈 0. 01 ）. The fluorescence intensity of fluo 3/AM was （ 169 ± 4） %, （ 169± 4） %, （220 ± 7） % in the OGD, Aβ and OGD ＋Aβgroups respectively, significantly higher than that in the control group （ P 〈 0.05 or P 〈 0.01 ）. Conclusion It showed that OGD ＋Aβ caused SHSYSY cells injury was greater than the OGD and Aβ groups. The possible mechanisms seemed to enhancement the activity of p38MAPK, and then disturbed calcium homeostasis, result in the apoptosis of SHSYSY cells.