Zhang Yuan, Yu Qian, Lai Tian-Bao, Yang Yang, Li Gang, Sun Sheng-Gang
Department of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Department of Neurology, Shandong Qianfoshan Hospital, Jinan, 250014, China.
J Huazhong Univ Sci Technolog Med Sci. 2013 Jun;33(3):427-432. doi: 10.1007/s11596-013-1136-5. Epub 2013 Jun 17.
Alzheimer's disease (AD) is an age-related, progressive neurodegenerative disorder that occurs gradually and results in memory, behavior, and personality changes. Abnormal sphingolipid metabolism was reported in AD previously. This study aimed to investigate whether sphK1 could exacerbate the accumulation of amyloid protein (Aβ) and sharpen the learning and memory ability of the animal model of AD using siRNA interference. An adenovirus vector expressing small interfering RNA (siRNA) against the sphK1 gene (sphK1-siRNA) was designed, and the effects of sphK1-siRNA on the APP/PS1 mouse four weeks after treatment with sphK1-siRNA hippocampal injection were examined. SphK1 protein expression was confirmed by using Western blotting and ceramide content coupled with S1P secretion was evaluated by enzyme-linked immunosorbent assay (ELISA). Aβ load was detected by immunohistochemical staining and ELISA. Morris water maze was adopted to test the learning and memory ability of the APP/PS1 mice. A significant difference in the expression of sphK1 protein and mRNA was observed between the siRNA group and the control group. Aβ load in transfected mice was accelerated in vivo, with significant aggravation of the learning and memory ability. The sphK1 gene modulation in the Aβ load and the learning and memory ability in the animal model of AD may be important for the treatment of AD.
阿尔茨海默病（AD）是一种与年龄相关的进行性神经退行性疾病，其发病缓慢，会导致记忆、行为和性格改变。此前有报道称AD患者存在鞘氨醇激酶1（sphK1）异常代谢。本研究旨在通过小干扰RNA（siRNA）干扰来探究sphK1是否会加剧淀粉样蛋白（Aβ）的积累并损害AD动物模型的学习和记忆能力。设计了一种表达针对sphK1基因的小干扰RNA（siRNA）的腺病毒载体（sphK1-siRNA），并检测了海马注射sphK1-siRNA四周后其对APP/PS1小鼠的影响。通过蛋白质免疫印迹法确认sphK1蛋白表达，并采用酶联免疫吸附测定（ELISA）评估神经酰胺含量及S1P分泌情况。通过免疫组织化学染色和ELISA检测Aβ含量。采用莫里斯水迷宫试验检测APP/PS1小鼠的学习和记忆能力。siRNA组与对照组之间sphK1蛋白和mRNA表达存在显著差异。转染小鼠体内Aβ含量增加，学习和记忆能力显著受损。AD动物模型中sphK1基因调控对Aβ含量及学习和记忆能力的影响可能对AD治疗具有重要意义。
