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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 To investigate the protective effect of ethyl acetate extract of Actinidia on dopaminergic neurons induced by 1-methyl-4-phenylpyridinium ion (MPP +). Methods The SH-SY5Y cell line was used to determine the half-lethal dose (IC50) at 48 h after MPP + incubation. Then, the cells were preincubated for 4 h with 25 mg / L, 50 mg / L, 100 mg / L isocratic ethyl acetate extracts and then incubated with 1.0 mmol / L (IC50 value) MPP + for 48 h. Cell viability was measured by MTT assay; apoptosis was observed by flow cytometry with Hoechst staining and AnnexinⅤ-FITC / PI double staining. Detection of intracellular reactive oxygen species (ROS) by ethyl acetate extract of D. nigra by DCFHDA. Results When the concentration of 1.0mmol / L MPP + was incubated for 48 hours, the lethal dose was reached, and the concentration of MPP + was 1.0mmol / L. Preincubation with ethyl acetate acetate (50mg / L) could significantly increase the viability of SH-SY5Y cells and decrease the rate of apoptosis. After the injury of MPP +, the level of ROS increased sharply, while the ROS level decreased after the intervention with 25mg / L, 50mg / L and 100mg / L ethyl acetate extract. Conclusion Ethyl acetate phase of 50mg / L Dictyosoma nigrum can significantly reduce the toxic injury of MPP + to SH-SY5Y cells and has a significant protective effect.