EX527对热量限制保护SH-SY5Y细胞的影响  

Effect of EX527 on the protection of SH-SY5Y by calorie restriction

作　　者：张静爽 马丽娜[1,2] 王蓉[1,2,3,4] 张景燕[1,2,4] 吴燕川[1,2,4] 赵志炜[1,2,4] 王玉兰[1,2,4] 

机构地区：[1]首都医科大学宣武医院中心实验室,北京100053 [2]北京市老年病医疗研究中心,北京100053 [3]北京脑重大疾病研究院阿尔茨海默病研究所,北京100053 [4]神经变性病教育部重点实验室,北京100053

出　　处：《首都医科大学学报》2015年第5期705-708,共4页Journal of Capital Medical University

基　　金：国家自然科学基金(81472007);北京市自然科学基金(7132044)~~

摘　　要：目的观察SIRT1抑制剂EX527对热量限制处理的人神经母细胞瘤株SH-SY5Y细胞的影响。方法体外培养SHSY5Y细胞,分为正常对照组(NC组)、热量限制组(CR组)和热量限制加SIRT1抑制剂组(CR+EX527组),光镜下观察细胞形态学改变,测定各组细胞噻唑蓝(thiazolyl blue tetrazolium bromide,MTT)代谢率及乳酸脱氢酶(lactate dehydrogenase,LDH)漏出率,应用蛋白免疫印迹(Western blotting)法测定SIRT1表达量。结果与NC组相比,CR组细胞形态好,表现为细胞胞体饱满、突起伸展、贴壁牢固,MTT代谢率升高(P<0.05),LDH漏出率有降低趋势,CR组和CR+EX527组SIRT1表达均增加(P<0.05);与CR组相比,CR+EX527组细胞形态较差,突起回缩,贴壁性不好,MTT代谢率降低(P<0.05),LDH漏出率有升高趋势,SIRT1表达略减少。结论热量限制对神经细胞具有保护作用,而SIRT1抑制剂EX527能够减弱热量限制的神经保护作用,提示热量限制对SH-SY5Y细胞的保护作用可能是由SIRT1介导的。Objective To observe the impact of SIRT1 inhibitor（ EX527） on the human neuroblastoma cell line SH-SY5 Y,and the SH-SY5 Y cells were treated with calorie restriction. Methods SH-SY5 Y cells were cultured in vitro and divided into the following three groups： normal control group（ NC group）,calorie restriction group（ CR group） and calorie restriction plus SIRT1 inhibitor EX527 group（ CR ＋ EX527 group）. The morphological changes in the SH-SY5 Y cells of each group were observed by light microscopy,and the thiazolyl blue tetrazolium bromide（ MTT） metabolic rate and the lactate dehydrogenase（ LDH） leakage rate of all groups were measured by MTT working liquid and LDH Cytotoxicity Assay Kit,and the expression of SIRT1 protein in the three groups mentioned above were detected by Western blotting. Results Compared with the normal control（ NC） group,cells of CR group were in good shape,including plump cell bodies,good adherence and neurites extension. The MTT metabolic rate of CR group increased（ P〈0. 05）,while the LDH leakage rate tended to decrease as compared with NC group. And the expression of SIRT1 protein of CR group and CR ＋ EX527 group were increased obviously（ P〈0. 05） as compared with NC group. Compared with the calorie restriction（ CR） group,cells of CR ＋EX527 group showed poor morphology,including no plump cell bodies,retracted neurites and bad adherence. The MTT metabolic rate of CR ＋ EX527 group decreased（ P〈0. 05）,while the LDH leakage rate tended to increase as compared with CR group. And the expression of SIRT1 of CR ＋ EX527 group were decreased slightly as compared with CR group. Conclusion Calorie restriction has a protective effect on nerve cells,and SIRT1 inhibitor EX527 can weaken the neuroprotective effect of calorie restriction in this study,so we preliminarily infer that the protective effect of calorie restriction on SH-SY5 Y cells may be mediated by SIRT1.

关 键 词：热量限制 SIRT1抑制剂 人神经母细胞瘤株SH-SY5Y 

分 类 号：R592[医药卫生—老年医学]