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纸质出版:2014
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卢茜,邓钰双,刘莉,张玉平,郭秀明,余刚.姜黄素对N2a/APP695细胞载脂蛋白E表达的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(01):46-49.
Lu Xi,Deng Yushuang,Liu Li,Zhang Yuping,Guo Xuming,Yu Gang.Curcumin suppresses ApoE expression in murine neuroblastoma Neuro2a/APP695 cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(01):46-49.
卢茜,邓钰双,刘莉,张玉平,郭秀明,余刚.姜黄素对N2a/APP695细胞载脂蛋白E表达的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(01):46-49. DOI:
Lu Xi,Deng Yushuang,Liu Li,Zhang Yuping,Guo Xuming,Yu Gang.Curcumin suppresses ApoE expression in murine neuroblastoma Neuro2a/APP695 cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(01):46-49. DOI:
目的 探讨姜黄素对N2a/APP695细胞载脂蛋白E(apolipoprotein E
ApoE)的影响及其可能机制。 方法 体外培养野生型的N2a细胞与阿尔茨海默病模型细胞,即N2a/APP695细胞;以MTT比色法检测不同浓度姜黄素对N2a/APP695细胞存活率的影响;应用5、10、20 μmol/L姜黄素处理细胞48 h后,通过RT-PCR、Western blot分别检测ApoE、低密度脂蛋白受体(low density lipoprotein receptor, LDLR)的mRNA、蛋白水平的变化,采用免疫细胞化学检测ApoE分布情况及表达变化。 结果 MTT检测结果显示
姜黄素对N2a/APP695细胞具有明显的保护作用,其中,20 μmol/L时保护作用最显著(
P
0.05)。RT-PCR、Western blot检测结果显示,与对照组相比,N2a/APP695细胞ApoE的mRNA及蛋白水平显著增加(
P
0.01),而LDLR水平明显减少(
P
0.01);姜黄素处理N2a/APP695细胞后,ApoE的mRNA及蛋白水平显著减少,LDLR的mRNA及蛋白水平明显增加,均呈剂量依赖性(
P
0.05)。免疫细胞化学检测结果显示ApoE主要分布在胞质、胞膜;与对照组相比,N2a/APP695细胞ApoE阳性染色数增多并浓染;经姜黄素处理后,ApoE阳性染色数减少并浅染(
P
0.01)。 结论 姜黄素可显著地抑制体外培养神经元中ApoE的产生,其机制可能与促进ApoE受体LDLR的表达有关。
Objective To determine the effect of curcumin on the expression of apolipoprotein E (ApoE) in murine neuroblastoma Neuro2a (N2a) cells. Methods MTT assay was used to detect the effect of curcumin at the doses of 2.5
5
10
20 and 40 μmol/L on the viability of N2a/APP695 cells
a cellular model of Alzheimer’s disease. After the N2a/APP695 cells then were treated w
ith curcumin at 5
10 and 20 μmol/L for 48 h
the mRNA levels of ApoE and low density lipoprotein receptor (LDLR) in N2a cells were evaluated by RT-PCR
and their protein levels were determined by Western blotting. The cellular content of ApoE was measured by immunocytochemical staining. Results Curcumin treatment resulted in significant increase in the cell viability
especially at the dose of 20 μmol/L (
P
0.05). Compared to normal cells
the mRNA and protein expression of ApoE were increased in N2a/APP695 cells
the LDLR expression
instead
were decreased (
P
0.01). Treatment with curcumin distinctly decreased ApoE expression and increased LDLR production in a does-dependent manner (
P
0.05). Further more
ApoE was mainly located in the cytoplasm and cell membrane
and the number of ApoE positive cells were increased and hyper-chromatic compared to normal cells. but the number of ApoE positive cells were decreased and the color grew pallid after the treatment of curcumin (
P
0.01). Conclusion Curcumin inhibits the ApoE production in N2a/APP695 cells via facilitating the expression of LDLR.
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