亚甲基蓝对冈田酸所致SK-N-SH细胞分化神经元损伤的保护作用

doi:10.7506/spkx1002-6630-201617033

食品科学

亚甲基蓝对冈田酸所致SK-N-SH细胞分化神经元损伤的保护作用

佀营营1，袁玉婷2，邱理红2，吕欢欢3，周慧2，许波1，李刚1，李忌1，王振华1,*

1.烟台大学生命科学学院，线粒体与健康衰老研究中心，山东烟台 264005；2.石河子大学药学院，新疆石河子 832005；
3.中国科学院西北高原生物研究所，青海西宁 810001

出版日期:2016-09-15 发布日期:2016-09-22

Protective Effect of Methylene Blue on Neuronal Damage Induced by Okadaic Acid in Differentiated SK-N-SH Neuroblastoma Cells

SI Yingying1, YUAN Yuting2, QIU Lihong2, LÜ Huanhuan3, ZHOU Hui2, XU Bo1, LI Gang1, LI Ji1, WANG Zhenhua1,*

1. Center of Mitochondria and Healthy Aging, College of Life Sciences, Yantai University, Yantai 264005, China;
2. School of Pharmacy, Shihezi University, Shihezi 832005, China; 3. Northwest Institute of Plateau Biology,
Chinese Academy of Sciences, Xining 810001, China

Online:2016-09-15 Published:2016-09-22

摘要/Abstract

摘要：

目的：考察亚甲基蓝（methylene blue，MB）对冈田酸（okadaic acid，OA）所致分化神经元损伤的保护作用，并探究其作用机制。方法：采用全反式维甲酸诱导SK-N-SH人神经母细胞瘤细胞分化为成熟神经元细胞；OA诱导分化神经元损伤模拟阿尔茨海默症（Alzheimer’s disease，AD）神经元损伤；磺酰罗丹明B法检测MB对神经元细胞活性的影响；Giemsa染色法观察细胞形态变化；Western blotting检测突触素蛋白、磷酸化tau蛋白（p-tau）的表达水平。结果：全反式维甲酸可浓度依赖性抑制SK-N-SH细胞增殖，10 μmol/L全反式维甲酸处理SK-N-SH细胞7 d后，细胞突触明显伸长，出现典型神经元特征；以40 nmol/L OA构建AD模型；MB可浓度依赖性抑制分化的SK-N-SH细胞活性，与OA组相比，随着MB浓度增加，细胞轴突长度与胞体长度比值增大，细胞突触变长，MB改善了OA所致的细胞突触损伤，突触素蛋白表达量明显升高（P＜0.01），p-tau（Ser262）蛋白水平明显降低（P＜0.01）。结论：MB对OA所造成的分化SK-N-SH神经元细胞突触损伤有保护作用，可防止AD中tau蛋白的过度磷酸化。

关键词: 亚甲基蓝, 分化SK-N-SH人神经母细胞瘤细胞, 全反式维甲酸, 冈田酸, tau蛋白磷酸化

Abstract:

Objective: To investigate the protective effect and underlying mechanism of methylene blue (MB) on neuronal
damage induced by okadaic acid (OA). Methods: The mature neurons differentiated from SK-N-SH neuroblastoma cells
induced by all-trans retinoic acid (ATRA) were exposed to OA to induce synaptic atrophy and cell injury. The SRB assay
was used to measure the effect of MB on the proliferation of differentiated SK-N-SH cells. Giemsa staining was used to
observe cell morphology. The expression of synaptophysin and the phosphorylation of tau protein were detected by Western
blotting. Results: ATRA inhibited the proliferation of SK-N-SH cells in a dose-dependent manner in vitro. The 10 μmol/L
ATRA exposure for 7 days could induce SK-N-SH neuroblastoma cells differentiation into mature neurons with obviously
extended synapse. Treatment with 40 nmol/L OA resulted in synaptic atrophy in differentiated SK-N-SH neuroblastoma
cells. MB inhibited the proliferation of differentiated SK-N-SH cells in a dose-dependent manner in vitro. Moreover,
MB treatment significantly increased the ratio of axon length to cell body length and the expression of synaptophysin
(P < 0.01), while it decreased the phosphorylation of tau protein at Ser262 site (P < 0.01). Conclusion: MB can alleviate
synaptic atrophy induced by OA, which may be due to the down-regulation of tau phosphorylation at Ser262 site.

Key words: methylene blue, differentiated SK-N-SH neuroblastoma cells, all-trans retinoic acid, okadaic acid, tau phosphorylation

中图分类号:

R966

佀营营，袁玉婷，邱理红，吕欢欢，周慧，许波，李刚，李忌，王振华. 亚甲基蓝对冈田酸所致SK-N-SH细胞分化神经元损伤的保护作用[J]. 食品科学, doi: 10.7506/spkx1002-6630-201617033.

SI Yingying, YUAN Yuting, QIU Lihong, Lü Huanhuan, ZHOU Hui, XU Bo, LI Gang, LI Ji, WANG Zhenhua. Protective Effect of Methylene Blue on Neuronal Damage Induced by Okadaic Acid in Differentiated SK-N-SH Neuroblastoma Cells[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201617033.

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亚甲基蓝对冈田酸所致SK-N-SH细胞分化神经元损伤的保护作用

Protective Effect of Methylene Blue on Neuronal Damage Induced by Okadaic Acid in Differentiated SK-N-SH Neuroblastoma Cells

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