Department of Psychiatry, Institute of Psychiatric Research, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
PLoS One. 2011;6(7):e21954. doi: 10.1371/journal.pone.0021954. Epub 2011 Jul 22.
Overproduction of amyloid-β (Aβ) protein in the brain has been hypothesized as the primary toxic insult that, via numerous mechanisms, produces cognitive deficits in Alzheimer's disease (AD). Cholinesterase inhibition is a primary strategy for treatment of AD, and specific compounds of this class have previously been demonstrated to influence Aβ precursor protein (APP) processing and Aβ production. However, little information is available on the effects of rivastigmine, a dual acetylcholinesterase and butyrylcholinesterase inhibitor, on APP processing. As this drug is currently used to treat AD, characterization of its various activities is important to optimize its clinical utility. We have previously shown that rivastigmine can preserve or enhance neuronal and synaptic terminal markers in degenerating primary embryonic cerebrocortical cultures. Given previous reports on the effects of APP and Aβ on synapses, regulation of APP processing represents a plausible mechanism for the synaptic effects of rivastigmine. To test this hypothesis, we treated degenerating primary cultures with rivastigmine and measured secreted APP (sAPP) and Aβ. Rivastigmine treatment increased metabolic activity in these cultured cells, and elevated APP secretion. Analysis of the two major forms of APP secreted by these cultures, attributed to neurons or glia based on molecular weight showed that rivastigmine treatment significantly increased neuronal relative to glial secreted APP. Furthermore, rivastigmine treatment increased α-secretase cleaved sAPPα and decreased Aβ secretion, suggesting a therapeutic mechanism wherein rivastigmine alters the relative activities of the secretase pathways. Assessment of sAPP levels in rodent CSF following once daily rivastigmine administration for 21 days confirmed that elevated levels of APP in cell culture translated in vivo. Taken together, rivastigmine treatment enhances neuronal sAPP and shifts APP processing toward the α-secretase pathway in degenerating neuronal cultures, which mirrors the trend of synaptic proteins, and metabolic activity.
脑内淀粉样-β(Aβ)蛋白的过度产生被假设为主要的毒性损伤,通过多种机制导致阿尔茨海默病(AD)的认知缺陷。胆碱酯酶抑制是治疗 AD 的主要策略,该类别的特定化合物先前已被证明可影响 Aβ前体蛋白(APP)的加工和 Aβ的产生。然而,关于rivastigmine(一种乙酰胆碱酯酶和丁酰胆碱酯酶的双重抑制剂)对 APP 加工的影响的信息很少。由于该药物目前用于治疗 AD,因此对其各种活性进行表征对于优化其临床应用非常重要。我们之前已经表明,rivastigmine 可以保留或增强退化的原代胚胎大脑皮质培养物中的神经元和突触末端标志物。鉴于先前关于 APP 和 Aβ对突触的影响的报告,APP 加工的调节代表了 rivastigmine 对突触作用的合理机制。为了验证这一假设,我们用 rivastigmine 处理退化的原代培养物,并测量分泌的 APP(sAPP)和 Aβ。rivastigmine 处理增加了这些培养细胞的代谢活性,并增加了 APP 的分泌。对这些培养物分泌的两种主要形式的 APP 进行分析,根据分子量归因于神经元或神经胶质,结果表明,rivastigmine 处理显著增加了神经元相对于神经胶质分泌的 APP。此外,rivastigmine 处理增加了α-分泌酶切割的 sAPPα,并减少了 Aβ的分泌,表明 rivastigmine 通过改变分泌酶途径的相对活性来改变治疗机制。在每日一次给予 rivastigmine 治疗 21 天后,评估啮齿动物 CSF 中的 sAPP 水平证实,细胞培养物中 APP 水平的升高在体内得到了证实。总之,rivastigmine 处理增强了神经元的 sAPP,并使 APP 加工向退化的神经元培养物中的α-分泌酶途径转移,这与突触蛋白和代谢活性的趋势一致。