乙醇对人神经母细胞瘤细胞凋亡和半胱氨酸蛋白酶-3活性的影响  

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

作　　者：蒲蜀湘[1] 饶从磊[1] 梁兵[1] 袁芳[1] 吴培基[1] 钟真真[1] 高聪[1] 

机构地区：[1]广州医学院第二附属医院神经内科,510260

出　　处：《临床神经病学杂志》2013年第1期38-40,共3页Journal of Clinical Neurology

摘　　要：目的探讨乙醇对人神经母细胞瘤(SH-SY5Y)细胞调亡和半胱氨酸蛋白酶(caspase)-3活性的影响。方法使用不同浓度乙醇(50、100、200、400和800 mmol/L)处理SH-SY5Y细胞24 h,噻唑蓝比色法检测乙醇对细胞增殖的抑制作用;Hoechst33342染色观察细胞凋亡的形态学变化;比色法检测细胞caspase-3相对活性。结果与空白对照组比较,乙醇50 mmol/L组、100 mmol/L组、200 mmol/L组、400 mmol/L组和800mmol/L组的吸光度值(A570)显著降低(P<0.05～0.01),SH-SY5Y细胞凋亡率及caspase-3活性显著增高(均P<0.01);各组间比较,差异亦有统计学意义(均P<0.001)。空白对照组SH-SY5Y细胞核较大,染色质均匀;50 mmol/L乙醇组细胞核有少许固缩,少量染色质边聚、浓缩;100 mmol/L乙醇组致密的颗粒状或条状强蓝色荧光的细胞核轻度增多;200 mmol/L乙醇组细胞核固缩增加,染色质边聚、浓染;400 mmol/L乙醇组细胞核固缩明显增加,染色质边聚、致密浓染,出现核碎裂和凋亡小体;800 mmol/L乙醇组大量细胞凋亡,碎裂的细胞核增多,出现较多的凋亡小体。结论乙醇呈剂量依赖性诱导SH-SY5Y细胞凋亡,其机制可能与caspase-3的活性增高有关。Objective To investigate the effect of ethanol on the apoptosis and caspase-3 activity of human neuroblastoma （SH-SYSY） 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 MTY 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 retool/L, 200 mmol/L, 400 mmoL/L and 800 mmol/L groups were significantly reduced （P 〈 0. 05 -0.01 ） ; the apoptosis rate of SH-SYSY cells and the activity of caspase-3 were significantly increased （ all P 〈 0.01 ）. There were statistical significance amang each group （ all P 〈 0. 001 ）. The nuclei of SH-S^SY 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-SYSY cells.

关 键 词：乙醇 SH-SY5Y细胞 CASPASE-3 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]