| 杜怡峰,闫鹏,韩晓娟.Alzheimer病胰岛素信号传导通路的研究[J].济宁医学院学报,2012,(2):84-92 |
| Alzheimer病胰岛素信号传导通路的研究 |
| Study on the impairment of insulin signal transduction pathway in Alzheimer's disease |
| 投稿时间：2012-04-02 |
| DOI：10.3969/j.issn.10009760.2012.02.002 |
| 中文关键词: 阿尔茨海默病;β淀粉样蛋白;神经元;胰岛素信号转导通路;大鼠 |
| 英文关键词: Alzheimer's Disease;Amyloid protein;Neuron;Insulin signaling transduction pathway;Rats |
| 基金项目:山东省自然科学基金项目(Y2008C116) |
| 作者 | 单位 | | 杜怡峰 | 山东省立医院神经内科, 山东济南 250021 | | 闫鹏 | 山东省立医院神经内科, 山东济南 250021 | | 韩晓娟 | 山东省立医院神经内科, 山东济南 250021 |
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| 中文摘要: |
| 目的 探讨胰岛素信号传导通路相关蛋白变化在Alzheimer病发生发展中的作用。方法 1)体外原代培养Wistar胎鼠基底前脑胆碱能神经元并鉴定,以不同终浓度的纤维化Aβ1-40分别对原代培养的神经细胞进行干预,荧光显微镜观察细胞形态学变化,四甲基偶氮唑盐(MTT)法检测细胞活性,提取细胞总蛋白行免疫印迹(Western Blotting)检测神经元胰岛素信号转导通路相关蛋白的表达。2)用微量注射器在痴呆模型组大鼠侧脑室注射Aβ1-42寡聚体5μl,同样的方法于盐水(NS)组大鼠注射生理盐水5μl,正常组大鼠不做任何处理。2周后通过Y-迷宫实验检测大鼠行为学改变并取海马组织进行免疫组化染色。结果 1)以2μmol/L、5μmol/L和10μmol/L三个终浓度纤维化Aβ1-40进行干预的胆碱能神经元,细胞形态学观察及MTT实验均显示神经元活性有下降趋势,且下降程度与纤维化Aβ1-40浓度呈正相关。以μ5mol/L浓度的纤维化Aβ1-40对细胞进行不同时长(24h,48h,72h)干预后,细胞形态学观察及MTT实验均显示神经元活性的下降程度与纤维化Aβ1-40的干预时长呈正相关。2)以2μmol/L、5μmol/L和10μmol/L 3个终浓度纤维化Aβ1-40进行干预的胆碱能神经元,其胰岛素信号传导通路相关蛋白中,InsR(Insulin Receptor)、IRS-1[Insulin Receptor Substrate-IRS-1、Akt/PKB(serine/threonine protein kinase B)]及Bcl-2的表达减低,减低程度与纤维化Aβ1-40干预浓度呈正相关;以5mol/L终浓度的纤维化Aβ1-40对细胞进行不同时长(24h,48h,72h)干预后,InsR、IRS-1、Akt/PKB及Bcl-2表达的减低程度与纤维化Aβ1-40干预时长呈正相关。3)AD模型组大鼠海马区神经元InsR、IRS-1和Akt/PKB比对照组表达减低(P<0.05),差异有统计学意义。结论 Aβ可引起神经元损害和学习记忆功能障碍,其主要机制可能是介导了海马神经元胰岛素信号转导通路功能异常。 |
| 英文摘要: |
| Objective To explore the impact of fibrillar Aβ1-40 and soluble Aβ oligomers on the insulin signal transduction pathway in Alzheimer's disease(AD).Methods 1) Primitive rat basal forebrain neurons were cultured and evaluated with.Morphologic, cellular vigor and IR、IRS-1、Akt/PKB、CREB、Bcl-2 expressing changes were observed by fluorescent microscope, MTT assay, Western Blot respectively when the neurons were exposed to fibrillar Aβ1-40 with different ending concentrations(0.1μmol/L, 1.0μmol/L, 2.0μmol/L, 5.0μmol/L, 10.0μmol/L)and different duration.2)5μl soluble Aβ oligomers were injected into lateral ventriculus of the model group by a microinjector under the stereotaxic apparatus.5μl saline solution was injected into the NS group in the same way, and the control group received nothing.The behavior was evaluated by the Y-maze test after 2 weeks.And then the hippocampus was removed and underwent immunohistochemical staining.Results 1) After being exposed to fibrillar Aβ1-40 for 48h, the neurons exposed to higher ending concentration(2.0, 5.0 and 10.0μmol/L) of fibrillar Aβ1-40 showed significant changes in morphology and cellular vigor:average process quantity of the neurons, average process length and the length of the longest process of neurons were decreased significantly, and the results of MTT assay showed that average OD values were significantly decreased.2) After being exposed to the three higher concentrations(2.0, 5.0&10.0μmol/L) of fibrillar Aβ1-40, the expression of InsR、IRS-1、Akt/PKB and Bcl-2 were significantly decreased.When the neurons were cultured with 5 mol/L fibrillar Aβ1-40 for different duration(24h, 48h, 72h), the above indexes were decreased by time dependency.3)Compared with the control group, the expression of InsR, IRS-1, and Akt/PKB in AD group were decreased(P<0.05), which was statistically significant.Conclusion Conclusion downregulated insulin signaling pathway induced by fibrillar Aβ1-40 in cultured primitive rat basal fore-brain cholinergic neurons, which is dose-dependent, may contribute to the death and loss of cholinergic neurons in AD.Soluble Aβ oligomers could give rise to learning and memory disability.The mechanism may be that it lead to abnormity of the insulin signal transduction pathway of hippocampus neurons. |
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