美金刚对H_2O_2诱导人神经母细胞瘤SH-SY5Y细胞凋亡的影响及机制探讨

Influence of memantine on hydrogen peroxide-induced apoptosis of SH-SY5Y cells and its mechanism

目的观察美金刚(Mem)对H2O2诱导人神经母细胞瘤SH-SY5Y细胞凋亡的影响,并探讨其作用机制。方法取对数生长期的细胞,随机分为5组;A、B、C、D组分别加入终浓度为10、20、30μmol/L的Mem及等体积的PBS预处理2 h,A、B、C、E组再加入400μmol/L的H2O2继续培养24 h。MTT法测定细胞活性,Hoechst33342染色检测细胞凋亡,比色法检测Caspase-3相对酶活性。结果 A、B、C、D、E组细胞活性(A570值)分别为0.617 5±0.037 5、0.674 0±0.052 2、0.645 5±0.051 7、1.030 6±0.068 9、0.554 4±0.044 3,细胞凋亡率分别为35.98%±5.96%、31.48%±6.79%、32.28%±9.16%、2.04%±1.54%、45.24%±5.78%,Caspase-3相对酶活性分别为251.35±20.76、219.33±18.24、223.27±29.89、100.00±13.25、327.49±35.18,E组与D组比较,P均<0.01;A、B、C组与E组比较,P均<0.05。结论 Mem能够阻止H2O2诱导的SH-SY5Y细胞凋亡,其机制可能与抑制Caspase-3的活性有关。

Objective To observe the influence of memantine （Mere） on hydrogen peroxide-induced apoptosis of SH- SY5Y cells and to investigate its mechanism. Methods SH-SY5Y ceils in logarithmic growth phase were randomly divided into five groups. Cells in group A, B, C and D were respectively pretreated with a final concentration of 10, 20, 30μmol/ L of Mere and an equal volume of PBS for 2 h. Then, the cells in group A, B, C and E were cultured with 400 μmol/L of H202 for 24 h. The cell viability was detected by the MTT assay, the apoptosis was determined by Hoechst33342 staining and the caspase-3 relative activity was monitored by the colorimetric assay. Results The absorbance value A570 in group A, B, C, DandEwas0.6175±0.0375, 0.6740-±0.0522, 0.6455±0.0517, 1.0306±0.0689 and0.5544± 0.044 3 respectively. The apoptosis rate severally was 35. 98% ± 5. 96%, 31.48% ± 6. 79%, 32.28% ± 9. 16%, 2.04% ± 1.54% , 45.24% ± 5.78% and the relative activity of caspase-3 separately was 251.35 ± 20.76, 219.33 ± 18.24, 223.27 ± 29.89, 100.00± 13.25, 327.49 ± 35.18 in group A, B, C, D, E. When group E was compared with group D, the differences were statistically significant （ all P 〈 0.01 ）. When group A, B and C were compared with group E, the statistical significances were noted （all P 〈 0.05）. Conclusion Memantine can prevent the apoptosis induced by hydrogen peroxide in SH-SY5Y cells, which may be related to the inhibition of caspase-3 activity.