新型番荔枝酰胺衍生物FLZ对去血清培养损伤星形胶质细胞的保护作用

Protective effect of a novel squamosamide derivative FLZ against serum deprivation injury in rat primary cortical astrocytes

目的评价番荔枝酰胺衍生物FLZ对去血清培养损伤星形胶质细胞的作用,并探讨其可能的作用机制。方法在去血清培养的星形胶质细胞模型上,以3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞存活率,Hochest 33342染色观察细胞核形态变化,荧光比色法检测细胞内活性氧簇(ROS)生成。酶联免疫吸附测定(ELISA)方法检测细胞培养液中S100B蛋白含量。生化法检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活力及丙二醛(MDA)与谷胱甘肽(GSH)含量。结果去血清培养损伤星形胶质细胞使其存活率下降,细胞内ROS生成增多,S100B分泌增加,细胞内SOD活力下降,GSH-Px活力升高,MDA生成增多,GSH含量减少。FLZ能提高星形胶质细胞在去血清培养状态下的存活率,减少去血清培养引起的S100B过度分泌,减少细胞内ROS和MDA过度生成,减少GSH含量,抑制SOD活力下降,降低GSH-Px活力过度升高,从而降低去血清培养对星形胶质细胞引起的氧化应激损伤。结论 FLZ对星形胶质细胞去血清培养损伤有明显的保护作用,这种保护作用可能与降低去血清培养损伤引起的S100B分泌增加、减少细胞内ROS的生成及增强细胞自身抗氧化系统相关。

Objective To investigate the effect of a squamosamide derivative（compound FLZ） on serum deprivation injury in cultured rat primary cortical astrocytes.Methods Astrocytes were cultured under normal and serum-deprived condition.Firstly,the optimal concentrations of FLZ exerting protective effects against serum deprivation injury were determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay.Then,the morphological changes in cell nuclei were observed by Hochest 33342 staining under fluorescence microscope.The level of secreted S100B was determined by ELISA.The reactive oxygen species（ROS） in cultures were detected by fluorescence probe 2′,7′-dichlorodihydrofluorescin diacetate（DCFDA）.Finally,the activities of superoxide dismutase（SOD） and glutathione peroxidase（GSH-Px）,concentrations of malondialdehyde（MDA）and glutathione（GSH）were determined using biochemical methods.Results Serum deprivation could decrease the viability of astrocytes and increase intracellular ROS production and S100B secretion.The injuries also included the decrease in SOD activity and GSH concentration,and the increase in GSH-Px activity and MDA concentration.After treated with FLZ,the decrease in cell viability,over-secretion of S100B,over production of cellular ROS and MDA,SOD activity and GSH production attenuation,and elevation of GSH-Px activity were all inhibited.Conclusion FLZ demonstrated significantly protective effects against serum-deprivation in rat cortical astrocytes,and the possible mechanisms involved may be related with the inhibition of over-secreted of S100B,the attenuation of cellular ROS over-production and the enhancement of cellular anti-oxidative system.In conclusion,FLZ attenuates the oxidative stress induced by serum deprivation of cultured cortical astrocytes and thus enhances the cellular anti-oxidative system.