Influnces of ethanol on the apoptosis and caspase-3 activity of human neuroblastoma cells

Objective To investigate the effect of ethanol on the apoptosis and caspase-3 activity of human neuroblastoma(SH-SY5Y) cells.Methods SH-SY5Y cells were incubated with various concentrations(50,100,200,400,800 mmol/L) of ethanol for 24 h.The depressant effect by ethanol was determined by the MTT colorimetric assay.The morphology change in apoptotic cells were monitored by Hoechst33342 staining.The caspase-3 relative activity was assessed by the colorimetric assay.Results Compared with blank control group,the absorbance value(A570) in ethanol 50 mmol/L,100 mmol/L,200 mmol/L,400 mmol/L and 800 mmol/L groups were significantly reduced(P0.05-0.01);the apoptosis rate of SH-SY5Y cells and the activity of caspase-3 were significantly increased(all P0.01).There were statistical significance amang each group(all P0.001).The nuclei of SH-SY5Y cells were larger and the chromatin was uniform in the blank control group.However,there have been few pyknotic nuclei,little margination and condensation of nuclear chromatin in ethanol 50 mmol/L group;slightly increased dense granular or strip strong blue fluorescence nuclei in ethanol 100 mmol/L group;pyknotic nuclei increase,the margination and staining of nuclear chromatin in ethanol 200 mmol/L group;obviously augmented pyknotic nuclei,the margination and dense staining of nuclear chromatin,nuclear fragmentation and apoptotic bodies in ethanol 400 mmol/L group;a large number of apoptosis cells,nuclear fragmentation increase and more apoptotic bodies in ethanol 800 mmol/L group.Conclusion Ethanol-induced apoptosis shows a dose-dependent manner and its mechanism may be related to the elevation of caspase-3 activity in SH-SY5Y cells.