热量限制对SH-SY5Y细胞氧化损伤的影响  

Effect of Caloric Restriction on Oxidative Damage in Human Neuroblastoma Cell Line SH-SY5Y Cells

作　　者：陈娟[1,2] 张景燕[1,2] 王蓉[1,2] 赵静姝[1,2] 郭瑾[3] 吴燕川[1,2] 赵志炜[1,2] 

机构地区：[1]首都医科大学宣武医院中心实验室 [2]北京老年病医疗研究中心,神经变性病教育部重点实验室,北京100053 [3]中国人民解放军总医院301超声诊断科

出　　处：《中国比较医学杂志》2012年第10期33-36,I0001,共5页Chinese Journal of Comparative Medicine

基　　金：北京市自然科学基金(7082043);首都卫生发展科研专项项目(2011-1001-02);北京市卫生系统高层次卫生技术人才计划资助项目(2009-3-64)

摘　　要：目的观察热量限制培养条件下,SH-SY5Y细胞抗氧化应激损伤的能力。方法建立过氧化氢诱导的SH-SY5Y细胞损伤模型。体外培养SH-SY5Y细胞,分为对照组、损伤组(50、100、250、500、1 000μmol/L H2O2)、低糖组(2 g/L)、低糖+损伤组,进行细胞形态观察、测定各组细胞的噻唑蓝(MTT)代谢率、乳酸脱氢酶(LDH)漏出率。结果与对照组比较,(50、100、250、500、1 000)μmol/L H2O2损伤1 h后MTT代谢率测定细胞活力,50μmol/L组与对照组比较差异无统计学意义(P>0.05);其他组与对照组比较,随着H2O2浓度的增加,细胞活力呈递减趋势,差异具有显著性(P<0.01);选定250μmol/L H2O2组为损伤应激源。用低糖预处理细胞24 h,给与250μmol/L H2O2损伤1 h后测定MTT代谢率显示,与对照组比较,损伤组活力明显下降,低糖组活力上升(P<0.01);与损伤组比较,低糖+损伤组活力明显上升(p<0.01);继续培养至7 h发现,与对照组比较,低糖组活力上升(P<0.01);与损伤组比较,低糖+损伤组活力明显上升(P<0.01)。进一步检测LDH漏出率显示,损伤1 h后结果显示,与对照组比较,损伤组漏出率明显增加(P<0.05),低糖组漏出率稍有减少(P>0.05);与损伤组比较,低糖+损伤组漏出率明显减少(P<0.01);继续培养7h显示,低糖7h组与低糖1 h组比较,漏出稍有增多(P>0.05),低糖+损伤组7 h组与低糖+损伤组1 h比较漏出率稍有增加(P<0.05);细胞形态学观察显示,未加损伤之前,低糖组的细胞形态,与对照组比较无明显改变。加入损伤药物1h后的细胞形态与对照组比较无明显改变。加入损伤药物7 h后的细胞形态,低糖组和对照组细胞突起伸展良好细长,损伤组可见细胞数目明显减少,死细胞多,突起回缩,细胞明显变圆,贴壁性不好,透光性差。结论热量限制能提高神经细胞的抗氧化应激能力,增加细胞生存率,降低死亡率。Objective To observe the effect of caloric restriction on oxidative damage in human neuroblastoma cell line SH-SY5Y cells in vitro.Methods To establish an in vitro model of H2O2-induced oxidative stress damage of SH-SY5Y cells.SH-SY5Y cells were cultured in vitro.The cells were divided into four groups： control group,H2O2（250 μmol/L） group,low glucose（2g/L） group,and low glucose ＋ H2O2 group.Cell morphology,thiazolyl blue（MTT） metabolism rate,and lactate dehydrogenase（LDH） leakage rate were measured to observe the cell growth status in different groups.Results Compared with the control group,the MTT metabolism rate in the cells treated with 50 μmol/L H2O2 for one hour was not significantly changed（P0.05）,but it was significantly decreased in the 100,250,500,1000 μmol/L H2O2-treated groups than that of control group（P0.01）,and there was a tendency that along with the increase of H2O2 concentration,the cell activity was increasingly decreased.So,the H2O2 concentration of 250 μmol/L was selected to generate oxidative stress.The MTT metabolism rate in the cells pretreated with low glucose for 24 hours and treated with 250 μmol/L H2O2 for one hour was significantly decreased than that of control group（P0.01）,but significantly increased than that of the H2O2 group（P0.01）.When the cells were further cultured for 7 hours,the metabolism rate of the low glucose group was significantly higher than that of the control group（P0.01）.Compared with the H2O2 group,the metabolism rate of the low glucose ＋ H2O2 group was significantly increased（P0.01）.The LDH leakage rate in the cells treated with 250 μmol/L H2O2 for 1 hour was significantly increased than that of the control group（P0.05）.The LDH leakage rate in the low glucose treated group was slightly decreased,and the rate of the low glucose ＋ H2O2 group was significantly lower than that of the H2O2-treated group（P0.01）.When the cells were further culture for 7 hours,the LDH leakage rate of the low glucose group trea

关 键 词：氧化应激 SH-SY5Y细胞 细胞培养 热量限制 

分 类 号：R33[医药卫生—人体生理学] R734[医药卫生—基础医学]