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
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篇名: 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
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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、100 μmol/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、30 μmol/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10~40 μmol/L浓度范围内对C6细胞存活率无明显影响,大于40 μmol/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。
ABSTRACT: 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 μmol/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.
期刊: 2018年第29卷第23期
作者: 刘晓惠,赵延欣,刘学源
AUTHORS: LIU Xiaohui,ZHAO Yanxin,LIU Xueyuan
关键字: 姜黄素;高糖;p53;大鼠神经胶质瘤细胞C6;突触素
KEYWORDS: Curcumin; High glucose; p53; Glioma cells C6 of rats; Synaptophysin
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