姜黄素对高糖培养下大鼠神经胶质瘤细胞C6中p53、Siah-1和突触素蛋白表达的影响  

Effects of Curcumin on the Expression of p53,Siah-1 and Synaptophysin in Glioma Cells C6 of Rats Cultured in High Glucose

作　　者：刘晓惠[1] 赵延欣[1] 刘学源[1] LIU Xiaohui;ZHAO Yanxin;LIU Xueyuan(Dept.of Neurology,Shanghai Tenth People＇s Hospital,Tongji University School of Medicine,Shanghai 200072,China)

机构地区：[1]同济大学医学院附属第十人民医院神经内科,上海200072

出　　处：《中国药房》2018年第23期3178-3183,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81771131;81571033);上海市科研计划项目(No.17411950100)

摘　　要：目的:体外研究姜黄素对高糖培养下大鼠神经胶质瘤细胞C6中抑癌基因p53、E3泛素连接酶Siah-1和认知相关蛋白突触素(Synaptophysin)表达的影响。方法:采用CCK-8法检测10、20、30、40、50、60、70、80、90、100μmol/L姜黄素在高糖培养下作用C6细胞24、48 h后的细胞存活率。采用Western blot法和实时荧光定量聚合酶链式反应(RT-PCR)法检测作用24 h后正常对照组(无任何处理)、高糖培养组和姜黄素低、高浓度组(高糖+10、30μmol/L姜黄素)细胞中p53、Siah-1、Synaptophysin蛋白及mRNA的表达情况;另同法检测高糖培养下小干扰RNA(siRNA)沉默C6细胞中p53基因后对Siah-1蛋白及mRNA表达的影响。结果:姜黄素在10～40μmol/L浓度范围内对C6细胞存活率无明显影响,大于40μmol/L浓度后呈浓度依赖性降低细胞存活率。与正常对照组比较,高糖培养组C6细胞中p53、Siah-1蛋白及mRNA表达增强,Synaptophysin蛋白及mRNA表达减弱,差异均有统计学意义(P<0.05);与高糖培养组比较,姜黄素低、高浓度组C6细胞中p53、Siah-1蛋白及mRNA表达减弱,Synaptophysin蛋白及mRNA表达增强,差异均有统计学意义(P<0.05)。高糖培养下,沉默p53基因后的C6细胞中Siah-1蛋白及mRNA表达均明显减弱(P<0.05)。结论:高糖条件下,姜黄素可通过下调C6细胞中p53、Siah-1表达,进而上调Synaptophysin表达。OBJECTIVE：To study the effects of curcumin on the expression of tumor suppressor gene p53,E3 ligase Siah-1 and cognitive associated protein Synaptophysin in glioma cells C6 of rats cultured in high glucose. METHODS：CCK-8 assay was used to detect survival rate of C6 cells after treated with 10,20,30,40,50,60,70,80,90,100 μmol/L curcumin for 24,48 h under high glucose environment. Western blot method and RT-PCR method were adopted to detect the protein and mRNA expressions of p53,Siah-1 and Synaptophysin in C6 cells of normal control group（without any treatment）,high glucose treatment group,curcumin low-concentration and high-concentration groups（high glucose＋10,30 μmol/L curcumin）after cultured for 24 h. Same method was used to determine the effects of P53 gene of C6 cells on protein and mRNA expression of Siah-1 after transfected with siRNA under high glucose environment. RESULTS：Curcumin had no significant effect on the survival rate of C6 cells in the range of 10-40 μ mol/L. The survival rate of C6 cells decreased significantly in a concentration-dependent manner when the concentration of curcumin was higher than 40 μmol/L. Compared with normal control group,the protein and mRNA expressions of p53 and Siah-1 in C6 cells were enhanced in high glucose treatment group,while the protein and mRNA expressions of Synaptophysin were decreased,with statistical significance（P〈0.05）. Compared with high glucose treatment group,the protein and mRNA expressions of p53 and Siah-1 in C6 cells were decreased in curcumin low-concentration and high-concentration groups, while the protein and mRNA expressions of Synaptophysin were enhanced,with statistical significance（P〈0.05）. Under treated with high glucose,the protein and mRNA expressions of Siah-1 in C6 cells were decreased significantly after silencing p53 gene （P〈0.05）. CONCLUSIONS：Under high glucose environment,curcumin can up-regulate the expression of Synaptophysin in C6 cells by down-regulating the expression of p53 and Siah-1.

关 键 词：姜黄素 高糖 P53 大鼠神经胶质瘤细胞C6 突触素 

分 类 号：R965[医药卫生—药理学]