阿尔茨海默病(AD)是一种以记忆丧失为特征的神经退行性疾病,认知障碍,痴呆的行为和心理症状.治疗神经退行性疾病的药物疗效有限反映了其复杂的病因和发病机制。一种新的体外模型可能有助于弥合现有的临床前动物模型和人体临床试验之间的差距。从而确定可在即将进行的临床试验中探索的有希望的治疗靶点.通过协助识别作用机制和潜在危险,体外测试还可以缩短翻译的时间和费用。
目标:由于这些因素,我们的目标是在短时间内建立一个强大的、信息丰富的AD细胞模型.通过在小化合物(PAFC-16和BetA)的帮助下触发MAPK和NF-κβ信号通路,分别,在小鼠小胶质细胞(SIM-A9)和神经母细胞神经2a(N2a)细胞系中。
结果:PAFC-16在72小时后在SIM-A9和N2a细胞系中以3.12nM至25nM的浓度引发活化作用。72小时后,在SIM-A9和N2a细胞系中以12.5nM至25nM的浓度激活NF-κβ途径。激活剂化学物质的组合为MEK1/2-ERK和NF-κβ提供了合适的激活。激活剂显著启动APP和MAPT基因表达,以及蛋白质APP的表达,β。淀粉样蛋白,tau,还有p-tau.靶向途径的激活导致显著的形态学变化。
结论:我们可以推断MEK1/2-ERK和NF-κβ途径,分别,被PAFC-16和BetA化学品直接激活。MEK1/2-ERK通路的激活导致APP基因的激活,进而激活β。淀粉样蛋白,这反过来又导致斑块。此外,NF-κβ激活导致MAPT基因的激活,导致Tau和p-Tau蛋白活化,最终导致缠结。这可以在短短三天内付诸实践,具有高水平的活性和稳定性,可传递给接下来的三代(亚培养),具有显著的形态学变化。在小胶质细胞和成神经细胞系中,我们成功地创建了一个新的AD细胞模型。
Alzheimer\'s disease (AD) is a neurodegenerative disease characterized by memory loss, cognitive impairment, and behavioral and psychological symptoms of dementia. The limited efficacy of drugs for the treatment of neurodegenerative diseases reflects their complex etiology and pathogenesis. A novel in vitro model may help to bridge the gap between existing preclinical animal models and human clinical trials, thus identifying promising therapeutic targets that can be explored in upcoming clinical trials. By assisting in the identification of the mechanism of action and potential dangers, in vitro testing can also shorten the time and expense of translation.
OBJECTIVE: As a result of these factors, our objective is to develop a powerful and informative cellular model of AD within a short period of time. Through triggering the MAPK and NF-κβ signaling pathways with the aid of small chemical compounds (PAF C-16 and BetA), respectively, in mouse microglial (SIM-A9) and neuroblast Neuro-2a (N2a) cell lines.
RESULTS: PAF C-16, initiated an activation effect at a concentration of 3.12 nM to 25 nM in the SIM-A9 and N2a cell lines after 72 h. BetA, activated the NF-κβ pathway with a concentration of 12.5 nM to 25 nM in the SIM-A9 and N2a cell lines after 72 h. The combination of the activator chemicals provided suitable activation for MEK1/2-ERK and NF-κβ in more than three subcultures. Activators significantly initiate APP and MAPT gene expression, as well as the expression of proteins APP, β. Amyloid, tau, and p-tau. The activation of the targeted pathways leads to significant morphological changes.
CONCLUSIONS: We can infer that the MEK1/2-ERK and NF-κβ pathways, respectively, are directly activated by the PAF C-16 and BetA chemicals. The activation of MEK1/2-ERK pathway results in the activation of the APP gene, which in turn activates the β. Amyloid protein, which in turn results in plaque. Furthermore, NF-κβ activation results in the activation of the MAPT gene, which leads to Tau and p-Tau protein activation, which ultimately results in tangles. This can be put into practice in just three days, with a high level of activity and stability that is passed down to the next three generations (subculture), with significant morphological changes. In microglial and neuroblast cell lines, we were successful in creating a novel AD-cell model.