基础医学与临床 ›› 2022, Vol. 42 ›› Issue (6): 927-932.doi: 10.16352/j.issn.1001-6325.2022.06.008

• 研究论文 • 上一篇    下一篇

冈田酸诱导人神经母细胞瘤细胞系SH-SY5Y损伤

路亚岚, 周澧, 韩云林, 王克维, 秦川*   

  1. 中国医学科学院 医学实验动物研究所 北京协和医学院 比较医学中心 国家卫生健康委员会人类比较医学重点实验室北京市人类重大疾病实验动物模型工程技术研究中心,北京 100021
  • 收稿日期:2021-05-24 修回日期:2022-03-28 出版日期:2022-06-05 发布日期:2022-06-02
  • 通讯作者: * qinchuan@pumc.edu.cn
  • 基金资助:
    中国医学科学院医学与健康科技创新工程(2019-12M-1-004)

Okadaic acid induces damage of human neuroblastoma cell line SH-SY5Y

LU Ya-lan, ZHOU Li, HAN Yun-lin, WANG Ke-wei, QIN Chuan*   

  1. Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences; Comparative Medicine Center, Peking Union Medical College; Key Laboratory of Human Disease Comparative Medicine, National Health Commission, Beijing Engineering Research Center for Experimental Animal Models of Human Critical Disease, Beijing 100021, China
  • Received:2021-05-24 Revised:2022-03-28 Online:2022-06-05 Published:2022-06-02
  • Contact: * qinchuan@pumc.edu.cn

摘要: 目的 探索冈田酸(OA)诱导神经元微管蛋白相关蛋白(Tau)异常磷酸化对人神经母细胞瘤细胞系(SH-SY5Y)损伤的影响。方法 将OA按照不同浓度(20和40 nmol/L)和/或时间(0、12、24、48和72 h)处理SH-SY5Y细胞;光学显微镜下观察细胞形态学;透射电子显微镜观察细胞超微结构;TUNEL法检测细胞凋亡;Western blot检测p-Tau和caspase3蛋白的表达;RT-qPCR检测细胞凋亡抑制家族蛋白(IAPs)的mRNA表达。结果 与对照组相比,OA显著增加SH-SY5Y细胞Tau蛋白磷酸化(P<0.001),促进细胞损伤凋亡(P<0.001),损伤线粒体结构,同时也可显著上调IAPs表达(P<0.05)。结论 OA诱导的SH-SY5Y细胞的Tau蛋白异常磷酸化可导致细胞损伤凋亡,抑制内源凋亡通路或上调IAPs可能作为阿尔茨海默病(AD)防治的重要靶标。

关键词: 阿尔茨海默病, 冈田酸, 神经元微管蛋白相关蛋白, 神经原纤维缠结

Abstract: Objective To investigate the effect of okadaic acid (OA)-induced phosphorylation of neuron micro-tubule associated protein tau (Tau) on the damage of human neuroblastoma SH-SY5Y cells. Methods SH-SY5Y cells were incubated with different concentrations (20 and 40 nmol/L) of OA for 0, 12, 24, 48 and 72 hours followed by microscopy for cell morphology.Cell ultrastructure was observed by transmission electron microscope and cell apoptosis was detected by TUNEL assay. The protein expression of p-Tau and caspase3 was analyzed by Western blot and mRNA level of inhibitor of apoptosis family proteins (IAPs) was determined by RT-qPCR. Results OA induced phosphorylation of Tau protein in SH-SY5Y cells as compared to the result from control goup (P<0.001), promoted cell damage and apoptosis (P<0.001).OA also injured mitochondrial structure in a concentration and time dependent manner. It can also up-regulate IAPs expression significantly (P<0.05). Conclusions The aberrant phosphorylation of Tau protein in SH-SY5Y cells induced by OA may lead to cell injury and apoptosis. Inhibition of endogenous apoptotic pathways or up-regulation of IAPs family proteins provides potential targets for AD prevention and treatment.

Key words: Alzheimer's disease, okadaic acid, Tau, neurofibrillary tangle

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