乳清蛋白酶解物抑制神经母细胞瘤细胞钙超载作用研究  

Effect of Whey Protein Hydrolysate on Calcium Overload in Neuroblastoma Cells

作　　者：朱勇[1] 毕方方[1] 李艳春 张国利 肖波[1] ZHU Yon BI Fang-fan LI Yan-chun ZHANG Guo-lr XIAO Bo(Department ofNeurology, Xiangya Hospital, Central South Universi（y, Changsha, Hunan, 410008, China Department of Neurology, the First Affiliated Hospital of Hunan Medical College, Huaihua, Hunan, 418000, China)

机构地区：[1]中南大学湘雅医院神经内科,湖南长沙410008 [2]湖南医药学院第一附属医院神经内科,湖南怀化418000

出　　处：《现代生物医学进展》2016年第35期6812-6816,共5页Progress in Modern Biomedicine

基　　金：2016年度湖南省医药卫生科研计划项目(B2016196)

摘　　要：目的:探讨乳清蛋白酶解物通过抑制过氧化氢,减轻过氧化氢对神经母细胞瘤细胞损害的机制。方法:将培育的人神经母细胞瘤细胞随机分为四份,对照组:未添加H_2O_2以及乳清蛋白酶解物的正常培养基中的细胞;过氧化氢组:神经母细胞瘤细胞+50μLH_2O_2,乳清蛋白观察组:神经母细胞瘤细胞+50μLH_2O_2+200μg/mL乳清蛋白酶解物;乳清蛋白对照组:神经母细胞瘤细胞+200μg/mL乳清蛋白酶解物,观察四组细胞存活和凋亡情况,以及细胞内钙离子和线粒体膜电位变化情况。结果:①细胞存活率对比上,过氧化氢组细胞存活率和凋亡率与其他三组相比,差异有统计学意义(F=24.62和32.82,P<0.01);乳清蛋白观察组细胞存活率和死亡率与过氧化氢组相比差异有统计学意义(t=7.65和9.82,P<0.05)。②过氧化氢组钙离子荧光强度最强,差异有统计学意义(Z=5.12,P<0.05);而对照组和乳清蛋白对照组荧光强度明显低于过氧化氢组和乳清蛋白组,差异有统计学意义(Z=5.57和4.82,P<0.05),乳清蛋白组荧光强度强于两对照组(Z=6.68,和4.15,P<0.05),弱于过氧化氢组(Z=6.12,P<0.05);③膜电位变化比较,正常对照组和乳清蛋白对照组荧光强度最强,提示膜电位变化幅度最小,过氧化氢组荧光强度最弱,提示膜电位变化幅度最大;乳清蛋白组荧光强度低于对照组而高于过氧化氢组,提示乳清蛋白酶解物能够阻止过氧化氢引起的线粒体膜电位改变。结论:乳清蛋白酶解物能够减少因过氧化氢诱导的神经母细胞瘤细胞内钙离子升高,减少线粒体内外膜电位差,从而抑制细胞凋亡。Objective： To explore the mechanisms of whey protein hydrolysatc reducing the damage of hydrogen peroxide on neu- roblastoma cells through inhibiting hydrogen peroxide. Methods： The human ueuroblastoma cells were randomly divided into four groups. The cells in control group were cultured in normal medium without the medium containing H2O2 and whey protein hydrolysate, the cells in hydrogen peroxide group were treated with 50 μL H2O2, and the cells in whey protein observation group were treated with 50 μL H2O2＋200 μg/m whey protein hydrolysatc, and the cells in whey protein control group were treated with 200 μg/mL whey protein hydrolysate. The survival and apoptosis of the four groups and the changes of intracellular calcium and mitochondrial membrane potential were observed. Results： （1） Compared with the other three groups, the differences for the survival rate and apoptosis rate of cells in the hydrogen peroxide group were statistically significant （F=24.62 and 32.82, P〈0.01）; The survival rate and apoptosis rate of cells in the whey protein observation group had significant differences compared with hydrogen peroxide group （t=7.65 and 9.82, P〈0.05）. （2）The fluorescence intensity of calcium ion in hydrogen peroxide group was the highest （Z=5.12, P〈0.05 ）, while the fluorescence intensity of the control group and the whey protein control group was the weakest （Z=5.57 and 4.82, P〈0.05）. The fluorescence intensity of the whey protein observation group was stronger than those of the two control groups （Z=6.68 and 4.15, P〈0.05 ）, which was weaker than that of the hydrogen peroxide group （Z=6.12, P〈0.05 ）. （3）The fluorescence intensity of the normal control group and the whey protein control group were the highest, which indicated that the change of membrane potential was the least. The fluorescence intensity of the hydrogen peroxide group was the weakest, which indicated that the change of membrane potential was the sharpest. The fluorescence intensity of

关 键 词：乳清蛋白酶解物 过氧化氢 人神经母细胞瘤细胞 钙超载 

分 类 号：R739.4[医药卫生—肿瘤]