色钉菇乙酸乙酯相对SH-SY5Y细胞的保护作用  

Protective effects of ethyl acetate extract from Chroogomphis rutilus on SH-SY5Y cells damaged by 1-methyl-4-phenyl pyridine ion

作　　者：李革[1] 姚辉 刘川[1] 高志国[1] 王立安[1] 管振龙[1] 王艳芹[1] 

机构地区：[1]河北师范大学生命科学学院,石家庄050024

出　　处：《解剖学报》2016年第3期330-336,共7页Acta Anatomica Sinica

基　　金：河北省科学技术研究与发展计划项目(14236903D);河北省自然科学基金(H2013205165);河北省高等学校科学技术研究项目(ZD2015012)

摘　　要：目的探讨色钉菇乙酸乙酯提取物对1-甲基-4-苯基吡啶离子(MPP+)诱导损伤的多巴胺能神经元的保护作用。方法以SH-SY5Y细胞系为对象,首先测定MPP+孵育48 h时的半致死量(IC50)。然后用25mg/L、50mg/L、100mg/L等剂量的乙酸乙酯提取物预先孵育4 h,随后加入1.0mmol/L(IC50值)MPP+孵育48 h。通过MTT法检测细胞活性;Hoechst染色法、AnnexinⅤ-FITC/PI双染法流式细胞术检测法观察细胞凋亡情况。利用DCFHDA检测色钉菇乙酸乙酯提取物对细胞内活性氧(ROS)水平的影响。结果 1.0mmol/L MPP+孵育48 h时达到半致死剂量,MPP+的损伤浓度确定为1.0mmol/L。预先经色钉菇乙酸乙酯相(50mg/L)孵育,可显著提高SH-SY5Y细胞的活力,降低细胞凋亡率。MPP+损伤后ROS水平急剧升高,而25mg/L、50mg/L、100mg/L乙酸乙酯提取物干预后ROS水平下降。结论 50mg/L色钉菇乙酸乙酯相可显著减轻MPP+对SH-SY5Y细胞的毒性损伤,具有显著的保护作用。Objective The aim of present study is to observe the protective effects of ethyl acetate extract from Chroogomphis rutilus on dopaminergic neurons damaged by 1-methyl-4-phenyl pyridine ion（ MPP＋）. Methods SH-SY5 Y cells（ dopaminergic cells）,a cell line of human neuroblastoma,were served as the experimental subject. We examined the IC50 of MPP＋at 48 hours. The cells were preincubated with ethyl acetate extract from Chroogomphis rutilus for 4 hours with25 mg / L,50 mg / L,100 mg / L respectively,then added MPP＋1. 0mmol / L（ IC50value） to incubate for 48 hours. MTT assay,Hoechst 33342 staining and flow cytometric assay were used to detect the survival or apoptosis of SH-SY5 Y cells.DCFH-DA was used to detect the reactive oxygen speciess（ ROS） level on the cells of ethyl acetate extract from Chroogomphis rutilus. Results 1. 0mmol / L MPP＋caused the IC50 of cells at 48 hour,so 1. 0mmol / L was used as the lesion concentration. Pretreatment with ethyl acetate extract from Chroogomphis rutilus（ 50 mg / L） significantly increased the cell viability,and reduced the apoptosis rate of cells. After MPP＋lesion,the ROS level increased significantly,and the preincubation with ethyl acetate extract from Chroogomphis rutilus reduced the ROS level induced by MPP＋. Conclusion Ethyl acetate extract from Chroogomphis rutilus can significantly protect dopaminergic cells damaged with MPP＋.

关 键 词：色钉菇乙酸乙酯相 多巴胺能神经元 四甲基偶氮唑盐 HOECHST33342 流式细胞术 

分 类 号：R338[医药卫生—人体生理学]