›› 2009, Vol. 4 ›› Issue (04): 280-283.

• 论著 • 上一篇    下一篇

蛋白激酶C抑制剂白屈菜红碱在缺氧缺糖所致SHSY5Y细胞损伤中的作用

邹良玉,褚晓凡,饶宜光,李刚,付学军,卢艺   

  1. 518020 广东省深圳市暨南大学医学院附属第二医院 深圳市人民医院神经内科
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-04-20 发布日期:2009-04-20
  • 通讯作者: 褚晓凡

Role of Chelerythrine Chloride, the Protein Kinase C Inhibitor, on the Cell Damage of SHSY5Y Neuronal Cells from Oxygen-glucose Deprivation

  • Received:1900-01-01 Revised:1900-01-01 Online:2009-04-20 Published:2009-04-20

摘要: 目的 观察蛋白激酶C(protein kinase C,PKC)抑制剂白屈菜红碱对体外缺氧缺糖(OGD)SHSY5Y细胞的保护作用,探讨PKC与脑缺血的关系及白屈菜红碱神经保护作用的机制。方法 体外培养SHSY5Y神经母细胞瘤株,分为假OGD,OGD及白屈菜红碱(0.01mmol/L)伴或不伴二乙基二硫代氨基甲酸钠(diethyldithiocarbamate,DDC)(100nmmol/L)处理组,以四甲基偶氮唑盐微量酶反应比色法(MTT法)检测SHSY5Y细胞活性,应用HEt和fluo-3/AM的荧光强度测定OGD后SHSY5Y细胞超氧化物阴离子和钙离子的浓度。结果 与假OGD组相比,OGD组细胞活性显著下降(P <0.01),白屈菜红碱能提高OGD后SHSY5Y细胞的活性(P <0.01),这种神经保护作用可被DDC阻断。白屈菜红碱降低OGD后HEt和fluo-3/AM荧光强度(P <0.05~0.01)。结论 白屈菜红碱通过激活SHSY5Y细胞超氧化物歧化酶,降低细胞内超氧阴离子的浓度,进而抑制细胞内钙超载,阻断PKC,从而达到保护SHSY5Y细胞对抗OGD的细胞损害的作用。

关键词: 白屈菜红碱; 缺氧; 蛋白激酶C; 钙; 超氧化物类

Abstract: Objective To investigate the neuroprotective effect of chelerythrine chloride on SHSY5Y neuronal cells against oxygen-glucose deprivation(OGD), and the mechanisms of the neuroprotective effect.Methods SHSY5Y neuroblastoma cells were exposed to OGD, chelerythrine chloride (0.01mol/L) alone or chelerythrine chloride together with diethyldithiocarbamate (100 nmol/L was administrated,cell viability was measured by the MTT assay, intracellular concentration of superoxide anion and calcium were evaluated via the fluorescence intensity of HEt and fluo-3/AM.Results Compared with the sham group, the MTT value decreased significantly in the OGD group (P <0.01), while after OGD, the MTT value was higher significantly in the chelerythrine chloride group than that in the OGD group (P <0.01), the neuroprotective effect of chelerythrine chloride wasblocked by administration of diethyldithiocarbamate; the fluorescence intensity of HEt (P <0.05)and fluo-3/AM (P <0.01) was decreased significantly in the chelerythrine chloride group.Conclusion Chelerythrine chloride protects SHSY5Y neuronal cells against OGD by activating the superoxide dismutase, then reducing the intracellular concentration of superoxide anion and calcium, thereafter inhibiting protein kinase C.

Key words: Chelerythrine chloride; Anoxia; Protein kinase C; Calcium; Superoxides