色钉菇乙酸乙酯相对SH-SY5Y细胞的保护作用

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。

Protective effects of ethyl acetate extract from Chroogomphis rutilus on SH-SY5Ycells damaged by 1-methyl-4-phenyl pyridine ion

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-SY5Y 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 with 25mg/L, 50mg/L, 100mg/L respectively, then added MPP⁺ 1.0mmol/L（IC₅₀ value） to incubate for 48 hours. MTT assay, Hoechst 33342 staining and flow cytometric assay were used to detect the survival or apoptosis of SH-SY5Y 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 (50mg/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⁺.