病理浓度甲醛的累积导致小鼠神经母瘤细胞活力及黏附能力下降（英文）

发布时间：2019-05-11 03:01

【摘要】：衰老相关的认知损伤,如阿尔茨海默病(AD),是一种缓慢、渐进、不可逆的神经退行性疾病.内源(尿)甲醛浓度与AD病人的认知损害程度呈正相关.但是,在以前研究中所采用的甲醛浓度,通常高于AD患者和老年人的病理和生理浓度.因此,研究长时程低浓度(病理浓度)甲醛对神经类细胞的影响,对于揭示甲醛在AD发生发展过程的作用是非常必要的.本文参照AD患者尿甲醛浓度,采用每隔24 h连续传代接种加甲醛(10μmol/L)的方式,模拟AD病人尿甲醛浓度,观察长时程甲醛的累积对小鼠神经母瘤细胞(N2a)和原代海马神经元的影响.通过高效液相色谱、细胞活力和乳酸脱氢酶释放检测显示,随着孵育时间的延长,培养基内甲醛浓度积累升高,N2a细胞生长受到抑制,在甲醛孵育的后期,细胞死亡显著增加.全息成像显示,长时程病理浓度甲醛的积累,使细胞厚度增加、面积减小、神经突受损、显著削弱细胞的黏附能力.同样条件下,小鼠原代海马神经元的一级神经突的数目显著降低,表明病理浓度甲醛的积累能够削弱神经元之间的连接.免疫印迹和免疫荧光染色显示,甲醛的累积可以使N2a细胞内Tau蛋白的181位苏氨酸(T181)和396位丝氨酸(S396)磷酸化水平显著上升,可能是甲醛累积导致的神经突减少和形态改变的因素之一.以上结果证明,病理浓度甲醛在细胞生存环境中的累积,可以导致神经细胞损伤,尤其是神经突的异常改变.本文为探索甲醛代谢失调与老年认知损害之间关系和机制提供了新的启示.
[Abstract]:Aging-related cognitive impairment, such as Alzheimer's disease (AD), is a slow, progressive, irreversible neurodegenerative disease. There was a positive correlation between the concentration of endogenous formaldehyde and the degree of cognitive impairment in patients with AD. However, the formaldehyde concentration used in previous studies is usually higher than the pathological and physiological concentrations in AD patients and the elderly. Therefore, it is necessary to study the effect of long-term low concentration (pathological concentration) formaldehyde on neural cells in order to reveal the role of formaldehyde in the occurrence and development of AD. According to the urine formaldehyde concentration of AD patients, the urine formaldehyde concentration of AD patients was simulated by continuous passage vaccination with formaldehyde (10 渭 mol / L) every 24 hours. To observe the effect of long-term formaldehyde accumulation on mouse neuroblastoma cells (N2A) and primary hippocampal neurons. The results of high performance liquid chromatography (HPLC), cell activity and lactic dehydrogenase release showed that with the prolongation of incubation time, the accumulation of formaldehyde in the culture medium increased, the growth of N2A cells was inhibited, and the cell death increased significantly at the later stage of formaldehyde incubation. Holographic imaging showed that the accumulation of formaldehyde in long-term pathological concentration increased the thickness of cells, decreased the area, damaged the nerve process, and significantly weakened the adhesion ability of cells. Under the same conditions, the number of primary hippocampal neurons in mice decreased significantly, indicating that the accumulation of formaldehyde in pathological concentration could weaken the connection between neurons. Western imprinting and immunofluorescence staining showed that the accumulation of formaldehyde could significantly increase the phosphorylation of Tau protein T181 and S396 in N2A cells. It may be one of the factors leading to the decrease of neuroprocesses and morphological changes caused by formaldehyde accumulation. These results show that the accumulation of formaldehyde in the cell survival environment can lead to nerve cell injury, especially the abnormal changes of nerve processes. This paper provides a new enlightenment for exploring the relationship and mechanism between formaldehyde metabolism disorder and cognitive impairment in the elderly.
【作者单位】：重庆医科大学基础医学院生物化学与分子生物学教研室;中国科学院生物物理研究所脑与认知国家重点实验室;
【基金】：supported by grants from National Basic Research Program of China(2012CB911004) The National Natural Science Foundation of China(31270868,31200601)~~
【分类号】：R749.16

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