美金刚对过氧化氢诱导多巴胺能神经元氧化应激损伤的保护作用

摘要

目的 探讨美金刚对过氧化氢(H2O2)诱导的多巴胺能神经元氧化应激损伤的保护作用.方法 用400μmol/L H2O2诱导人神经母细胞瘤细胞SH-SY5Y损伤建立多巴胺能神经元氧化应激损伤模型,随机分为美金刚10、20、30 μmol/L组和空白对照组,分别加入相应试剂(空白对照组加入等体积PBS)；倒置显微镜观察细胞形态,MTT法检测细胞活性,比色法测定LDH释放率.结果 10、20、30μmol/L美金刚干预24h后,SH-SY5Y细胞的吸光度值(A570)分别为0.936 5±0.033 2、0.974 9±0.030 0和0.948 7±0.033 6,空白对照组为0.924 7±0.039 8；组间比较无统计学差异(P =0.160).经10、20、30 μmol/L美金刚预处理2h后,SH-SY5Y细胞的存活率分别为(59.92 ±3.64)％、(65.40±5.07)％和(62.63±5.01)％,与H2O2处理组比较有统计学差异(P＜0.05或＜0.01).此外,10、20、30 μmol/L美金刚预处理组SH-SY5Y细胞的LDH释放率分别为(38.70±7.88)％、(34.10±6.75)％、(35.58±8.68)％,与H2O2处理组比较均有统计学差异(P＜0.05或＜0.01).结论 美金刚对H2O2诱导的多巴胺能神经元氧化应激损伤具有保护作用.%Objective To study the protective effect of memantine against hydrogen peroxide-induced oxidative stress in dopaminergic neurons. Methods The oxidative stress damage model of dopaminergic neurons was established by the cultured human neuroblastoma cells SH-SY5Y with 400 μmol/L of hydrogen peroxide in vitro. The cell morphology was observed using the inverted microscope. The cell viability was detected by the MTT assay. The release rate of LDH was determined by the colorimetric assay. Results After SH-SY5Y cells were incubated with 10, 20, 30 μmol/L of memantine for 24 h, the absorbance value (A570) was 0.936 5 ± 0.033 2, 0.974 9 ± 0.030 0 and 0. 948 7 ± 0.033 6, respectively. The absorbance value (A570 ) in the control group was 0.924 7 ± 0.039 8. Any comparison between two groups was no significant difference (P=0.160). The SH-SY5Y cells were pretreated with 10, 20, 30 (μmol/L of memantine for 2 h, survival rate of the three memantine groups respectively was (59.92 ±3.64)% ,(65.40 ±5.07)% and (62.63 ±5.01)% , as compared with that of the hydrogen peroxide-cultivated group, the differences were statistically significant (P <0.05 or < 0.01). Furthermore, the release rate of LDH in 10, 20, 30 μmol/L of memantine-pretreated groups was (38. 70 ± 7. 88) % , (34.10 ± 6. 75) % and ( 35. 58 ± 8. 68 ) % , respectively. Compared with that of hydrogen peroxide-fostered . group, statistical significances were noted all(P <0.05 or <0.01). Conclusion Memantine has protective effects against hydrogen peroxide-induced oxidative stress impairment in dopaminergic neurons.