咖啡酸苯乙酯对鱼藤酮诱导帕金森细胞模型的保护  

Caffeic acid phenethyl ester against cellular injuries in the rotenone-induced Parkinson's disease model

作　　者：邱实 李军果 邱倩[2] 陈辉 相子民[3] 

机构地区：[1]解放军陆军总医院,北京市100068 [2]解放军第155医院呼吸肾病科,河南省开封市475003 [3]解放军南京军区福州总医院骨一科,福建省福州市350025

出　　处：《中国组织工程研究》2016年第40期5979-5985,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81501024)~~

摘　　要：背景:咖啡酸苯乙酯可抑制大鼠脑损伤后的脂质过氧化和脑损伤,但是在帕金森细胞模型中,5-脂氧酶和白三烯的变化规律,以及咖啡酸苯乙酯是否通过抑制鱼藤酮诱导的5-脂氧酶和白三烯变化而发挥对帕金森样损伤的保护作用,仍有待进一步研究。目的:探讨咖啡酸苯乙酯对鱼藤酮诱导帕金森细胞模型的保护作用,同时观察5-脂氧酶是否参与帕金森细胞模型的损伤。方法:(1)取生长状态良好的PC12细胞,分5组培养,分别以0,0.01,0.1,1,10μmol/L鱼藤酮干预,24,48 h后,观察细胞形态及活性变化,选择最佳诱导帕金森细胞模型的鱼藤酮浓度;24 h后,Western法检测5-脂氧酶表达,ELISA法检测半胱氨酰白三烯生成释放量;(2)取生长状态良好的PC12细胞,分6组培养,以正常培养的细胞为空白对照,其余5分别加入0,0.01,0.1,1,10μmol/L咖啡酸苯乙酯预处理30 min,之后加入1μmol/L鱼藤酮,24 h后,检测细胞活性,ELISA法检测半胱氨酰白三烯生成释放量。结果与结论:(1)0.1-10μmol/L鱼藤酮可诱导PC12细胞损伤,1μmol/L鱼藤酮处理24 h后的细胞形态和细胞存活率变化适度明显,所以选择该条件作为鱼藤酮诱导PC12细胞帕金森样损伤的细胞模型;(2)鱼藤酮呈浓度依赖性增加5-脂氧酶的表达与半胱氨酰白三烯的生成释放;(3)1-10μmol/L咖啡酸苯乙酯可降低鱼藤酮诱导的PC12细胞半胱氨酰白三烯生成释放量增加,呈浓度依赖性抑制鱼藤酮诱导的PC12细胞存活率降低;(4)结果表明,5-脂氧酶参与鱼藤酮诱导帕金森细胞模型的损伤作用,咖啡酸苯乙酯对鱼藤酮诱导帕金森细胞模型的损伤有明显保护作用。BACKGROUND： Caffeic acid phenethyl ester（CAPE） can inhibit lipid peroxidation after rat brain injury. However, the trend of 5-lipoxygenaseis（5-LOX） and cysteinyl leukotrienes（Cys LTs） in model of Parkinson＇s disease, and whether CAPE protects against rotenone-induced cellular injuries by inhibiting the levels of 5-LOX and CysL Ts still need further research. OBJECTIVE： To investigate the protective effect of CAPE on the rotenone-induced Parkinson-like injury, and to determine whether 5-LOX involved. METHODS：（1） PC12 cells in good-growth were collected and divided into five groups cultured with different concentrations of rotenone（0, 0.01, 0.1, 1, 10 μmol/L）. 24 and 48 hours later, changes of cellular morphology and activity were observed to single out the optimum concentration of rotenone; at 24 hours, the levels of 5-LOX and Cys LTs were detected by western blotting and ELISA, respectively.（2） PC12 cells were pretreated with different concentrations of CAPE（0, 0.01, 0.1, 1, 10 μmol/L） for 30 minutes, and 1 μmol/L rotenone was then added. The other cells received no intervention as blank control group. Subsequently, the cell activity was detected, and the CysL Ts production was detected by ELISA at 24 hours. RESULTS AND CONCLUSION：（1） Rotenone（0.1-10 μmol/L） could induce PC12 cell injury with overt morphological and cell activity changes at 24 hours, especially the 1 μmol/L rotenone.（2） Rotenone also significantly increased the 5-LOX expression and Cys LTs production in a concentration-dependant manner.（3） CAPE（1-10 μmo/L） significantly attenuated rotenone-induced Cys LTs production and cell viability reduction in a concentration-dependant manner.（4） These results suggest that CAPE protects against PC12 cell injuries in the model rat with Parkinson＇s disease induced by rotenone involving 5-Lox.

关 键 词：实验动物 神经损伤与修复动物模型 咖啡酸苯乙酯 5-脂氧酶 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]