建立Aβ1-42介导新生鼠原代神经元退行性变细胞模型及虾青素作用研究

Study on Establishing a Cellular Model of Primary Neuronal Degeneration Induced by Aβ1-42 and Screening Astaxanthin

目的:建立低聚Aβ1-42(Oligomeric Aβ1-42,oAβ1-42)介导Sprague-Dawley(SD)大鼠乳鼠原代神经元退行性变的细胞模型,用于阿尔茨海默病发病机制、药物研发或潜在新药如虾青素(Astaxanthin,AST)对其的预防和治疗作用筛选研究。方法:用oAβ1-42介导来源于2-3d龄SD新生鼠大脑皮质的成熟原代神经元,以产生的氧化应激产物一氧化氮(NO)、炎症介质环氧化酶(COX-2),以及凋亡蛋白Caspase-3、Bax作为模型的生物标记物,与对照组或脂多糖(LPS)组比较,判断模型建立是否成功;同时用该模型对虾青素的作用进行研究。结果:在建立的新生鼠原代神经元退行性变细胞模型中,Aβ1-42介导神经元产生毒性反应,与对照组比较,NO明显上调(P<0.05),Caspase-3、Bax和COX-2蛋白的表达明显增加;该来源的原代神经元对LPS的反应比Aβ1-42敏感;抗氧化剂虾青素可下调二者的毒性反应。结论:oAβ1-42介导2-3d龄SD乳鼠神经元发生退行性变的细胞模型建立成功;强抗氧化剂虾青素有较好调节oAβ1-42或LPS神经退行性变毒性反应的作用。该模型可望用于阿尔茨海默病Aβ1-42介导神经元氧化应激和神经退行变等发病机理研究以及药物研发、新药筛选的研究。

Objective To establish a cellular model of neuronal degeneration induced by oligomeric Aβ1-42 via using the primary neurons derived from 2-3 d Sprague-Dawley(SD)rats,and to apply to study the onset mechanisms of Alzheimer’s disease(AD)and Screening studies on the prevention and treatment of AD or potential drug like Astaxanthin for preventing or treating AD with this model.Methods Compare with LPS,Oligomeric Aβ1-42 was applied to primary neurons from 2-3 d SD neonatal rats,and the toxic target molecules such as Nitric Oxide(NO),Caspase-3,Bax,and Cyclooxygenase(COX-2)as oxidative-stress response and neuronal degenerative events were observed.Compared with the control group or the lipopolysaccharide(LPV)group,judge whether the model was established successfully.Moreover,Astaxanthin,as an anti-oxidative agent,was tested in this model.Results In this model of degenerative toxicity of primary neurons mediated by Aβ1-42 or LPS;Compared with the control group the NO product induced by oAβ1-42 or LPS was up-regulated,significantly(P<0.05);and the expression of Caspase-3,Bax and COX-2 induced by Aβ1-42 or LPS were obviously up-regulated,The sources of primary neurons are more sensitive to LPS than Aβ1-42.Furthermore,all of above toxic molecules were down-regulated by Astaxanthin.Conclusion A neuronal degenerative cellular model,which was using primary neurons cultured from the cortex of 2-3 d SD rat neonatal pups,induced by oAβ1-42 SD rats has been successfully established.Also,it was confirmed the anti-oxidative stress,anti-apoptosis and anti-neurodegenerative effects of Astaxanthin on this model.It can be useful for the studies-in the neuropathogenesis of oxidative stress induced by Aβ1-42 in Alzheimer’s disease,or drug development as well as screening drugs.