Department of Neurology, Philipps-University Marburg, Marburg, Germany.
J Alzheimers Dis. 2011;26(1):69-80. doi: 10.3233/JAD-2011-110179.
Amyloid-β (Aβ) oligomer toxicity is a crucial factor in the development of Alzheimer's disease. Therefore, the aim of therapeutic research is to target the modification of secretase activity, increase Aβ degradation, reduce Aβ formation, and modulate Aβ-induced neuroinflammation. Recently, the p38 MAP kinase inhibitor CNI-1493 has been shown to reduce plaque load and has led to an improvement in memory performance in a transgenic mouse model. We examined the role of CNI-1493 in the microglial inflammatory response to Aβ using both a microglia cell line as well as primary microglia isolated from mesocortices. MTT assays were performed to quantify cell viability. FACS analysis was used to measure phagocytosis. We used ELISA to analyse cytokine concentrations in response to CNI-1493 treatment. Western-blot/Dot-blot techniques were used to show the interaction of CNI-1493 with Aβ-oligomers as well as to measure apoptosis in microglia cells. RT-PCR was used to analyze secretase expression, and secretase function was determined using fluorimetric assays. CNI-1493 is able to prevent oligomer formation as well as apoptosis in microglia. A significant reduction was found in the Aβ-induced release of IL-6 and TNF-α in the presence of CNI-1493. Phagocytosis is an essential Aβ removal mechanism and was enhanced by CNI-1493 in primary microglia. CNI-1493 also influenced the α-secretase product C83 with an increase in the treated cells, while a simultaneous reduction in Aβ secretion was also observed. We hypothesize that CNI-1493 not only reduces neuroinflammation and consequent neurodegeneration, but also leads to a shift in AβPP-processing towards the non-amyloidogenic pathway. Therefore, CNI-1493 is a promising candidate for the treatment of AD.
淀粉样蛋白-β（Aβ）寡聚物毒性是阿尔茨海默病发展的关键因素。因此，治疗研究的目的是靶向修饰分泌酶活性，增加 Aβ 降解，减少 Aβ 形成，并调节 Aβ 诱导的神经炎症。最近，p38 MAP 激酶抑制剂 CNI-1493 已被证明能减少斑块负荷，并在转基因小鼠模型中改善记忆表现。我们使用小胶质细胞系和从小鼠中分离的原代小胶质细胞研究了 CNI-1493 在 Aβ 诱导的小胶质细胞炎症反应中的作用。MTT 法测定细胞活力。FACS 分析用于测量吞噬作用。我们使用 ELISA 分析 CNI-1493 处理后细胞因子浓度。Western-blot/Dot-blot 技术用于显示 CNI-1493 与 Aβ 寡聚物的相互作用以及测量小胶质细胞凋亡。RT-PCR 用于分析分泌酶表达，荧光法测定分泌酶功能。CNI-1493 能够防止小胶质细胞中寡聚物形成和凋亡。在 CNI-1493 存在的情况下，发现 Aβ 诱导的 IL-6 和 TNF-α释放显著减少。吞噬作用是 Aβ 清除的重要机制，在原代小胶质细胞中，CNI-1493 增强了吞噬作用。CNI-1493 还影响 α-分泌酶产物 C83，在处理细胞中增加，同时也观察到 Aβ 分泌减少。我们假设 CNI-1493 不仅减少神经炎症和随后的神经退行性变，而且还导致 AβPP 加工向非淀粉样形成途径的转变。因此，CNI-1493 是治疗 AD 的有前途的候选药物。
