姜黄素影响N2a/APP695swe细胞中ABCA1表达的机制研究  

Curcumin up-regulates ABCA1 expression through inhibiting the activity of CaN in N2a/APP695swe cells

作　　者：许可[1] 向月[1] 孙洁芸[1] 张雄[1] 李昱[1] 

机构地区：[1]重庆医科大学神经科学研究中心、病理学教研室,重庆400016

出　　处：《重庆医科大学学报》2015年第5期677-681,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81271426)

摘　　要：目的:研究姜黄素(Curcumin)对N2a/APP695swe细胞钙调神经磷酸酶(Calcineurin,CaN)活性和三磷酸腺苷结合转运子A1(ATP-binding cassette transporter A1,ABCA1)表达的影响;探讨Curcumin影响N2a/APP695swe细胞中ABCA1表达的可能机制。方法:MTT法分别检测1、5、10、20、40μmol/L Curcumin或0.1、0.5、1、2、4μmol/L CaN活性抑制剂环孢素A(Cyclosporin A,Cs A)对N2a/APP695swe细胞存活率的影响;依据MTT筛选结果,分为正常对照组、模型组、Curcumin组:5μmol/L Curcumin处理24 h的N2a/APP695swe细胞、Cs A组:0.5μmol/L Cs A处理48 h的N2a/APP695swe细胞。酶底物显色法检测CaN活性,real-time PCR、Western blot检测ABCA1表达水平的变化。结果:与未处理组比较,5μmol/L Curcumin组细胞存活率[(110.495±4.005)%]最高(P=0.023);0.1、0.5、1、2、4μmol/L Cs A组细胞存活率依次为(105.532±4.292)%、(115.211±4.826)%、(76.317±4.475)%、(69.177±5.267)%、(54.687±3.912)%,与未处理组比较,0.5、1、2、4μmol/L Cs A组差异均有统计学意义(P值依次为0.001、0.000、0.000、0.000);酶底物显色法显示与正常对照组[(18.519±1.664)nmol/mg]比较,模型组CaN活性[(24.488±3.041)nmol/mg]增加(P=0.007),Curcumin组[(16.717±2.055)nmol/mg]、Cs A组[(4.928±1.232)nmol/mg]CaN活性明显受到抑制(P值依次为0.002、0.000);real-time PCR和Western blot显示Curcumin组ABCA1表达量(real-time PCR:1.988±0.355;Western blot:0.294±0.015)高于模型组(real-time PCR:1.000±0.000;Western blot:0.175±0.008),差异有统计学意义(real-time PCR:P=0.009;Western blot:P=0.000);与模型组比较,Cs A组(real-time PCR:4.000±0.464;Western blot:0.470±0.016)的ABCA1表达水平增加(real-time PCR:P=0.000;Western blot:P=0.000)。结论:Curcumin能上调N2a/APP695swe细胞中ABCA1的表达,其机理可能与抑制CaN活性有关。Objective：To investigate the effect of Curcumin on the activity of Calcineurin（CaN）and the expression of ATP-binding cassette transporter A1（ABCA1）in N2a/APP695 swe cells,and to explore the mechanism of the regulation of Curcumin on the expression of ABCA1 in N2a/APP695 swe cells. Methods：MTT assay was used to detect N2a/APP695 swe cell viabilities in different concentrations of Curcumin or the CaN activity inhabitor cyclosporin A（Cs A）treatment. According to the results determined by MTT,N2a/APP695 swe cells were co-cultured with 5 μmol/L Curcumin for 24 h,or 0.5 μmol/L Cs A for 48 h,then,the CaN activities were measured by enzyme substrate color metric methods. Western blot and real-time PCR were performed to evaluate the expression of ABCA1. Results：Cell survival rate was（110.495±4.005）%,the highest in 5 μmol/L Curcumin group（P=0.023）,followed by（105.532±4.292）%,（115.211 ±4.826）%,（76.317±4.475）%,（69.177±5.267）%,（54.687±3.912）%,respectively in 0.1,0.5,1,2,4 μmol/L Cs A groups. There were significant differences in cell survival rate between untreated group and 0.5,1,2,4 μmol/L Cs A groups（P =0.001,0.000,0.000,0.000）. Compared with that（18.519±1.664）nmol/mg of normal group,the CaN activity was increased in N2a/APP695 swe cells（24.488 ±3.041） nmol/mg（P =0.007）,CaN activities were distinctly decreased in Curcumin group（16.717±2.055）nmol/mg（P=0.002）and Cs A group（4.928±1.232）nmol/mg（P=0.000）. Furthermore,Western blot and real-time PCR showed that the expression of ABCA1 was higher in Curcumin group（real-time PCR：1.988±0.355;Western blot：0.294±0.015）than in the model group（real-time PCR：1.000;Western blot：0.175±0.008）（P=0.009 in real-time PCR and P=0.000 in Western blot）. Compared with that of model group,the expression of ABCA1 in Cs A group（real-time PCR：4.000±0.464;Western blot：0.470±0.016）was increased（both P=0.000 in real-time PCR and Western blot）. Conclusion：Curcumin may up-regul

关 键 词：阿尔茨海默病 钙调神经磷酸酶 三磷酸腺苷结合盒转运子A1 姜黄素 环孢素A 

分 类 号：Q256[生物学—细胞生物学] R741.02[医药卫生—神经病学与精神病学]