黑质注射LPS构建亚急性帕金森病模型的评价

Evaluation of subacute Parkinson’s disease model by substantia nigra injection LPS

目的向黑质部位连续注入脂多糖(lipopolysaccha-ride,LPS)建立亚急性帕金森病(Parkinson’s disease,PD)小鼠模型。方法将60只雄性C57BL/6小鼠随机分为3组,模型组向双侧黑质注射LPS(2.5μg/侧),溶剂组以相同方法注射生理盐水,空白组不给予任何处理。连续注射5 d后,观察小鼠PD样行为学变化。免疫组化法观察黑质致密部多巴胺(DA)能神经元及活化小胶质细胞数目的变化,实时荧光定量PCR检测小鼠黑质内炎症因子的含量和West-ern blot观察不同形式α-突触核蛋白(α-synuclein,α-Syn)的变化。结果与溶剂组相比,模型组小鼠出现明显的PD样行为表现。与空白组相比,溶剂未对小鼠产生行为学障碍。模型组黑质致密部的DA能神经元显著减少,活化小胶质细胞显著增多,中脑内炎症因子mRNA的表达显著上调和三种形式的α-Syn的蛋白表达显著增加。结论经双侧黑质连续5 d注入LPS可诱导较为稳定的亚急性PD小鼠模型。

Aim To establish the model of subacute Parkinson’s disease(PD)by continuous injection of lipopolysaccharide(LPS)into the substantia nigra.Methods Sixty male C57BL/6 mice were randomly divided into three groups.LPS(2.5μg/side)was injected into bilateral substantia nigra in model group,saline was injected into the solvent group in the same way,and no treatment was given to the blank group.After 5 days of con-tinuous injection,PD like behavior of mice was observed.Im-munohistochemistry was used to observe the changes of dopamin-ergic neurons and activated microglia in mouse substantia nigra pars compacta.Real-time quantitative PCR(qPCR)was used to detect the content of inflammatory factors in mouse substantia nigra.Western blotwas used to observe different forms ofα-synu-clein.Results Compared with vehicle group,model group mice showed significant PD-like behavior.The solvent did not cause a behavioral disorder in mice compared to control group.The number of dopaminergic neurons in the substantia nigra pars compacta of the model group was significantly reduced,the num-ber of activated microglia was significantly increased,and the expression of inflammatory factor mRNA in the midbrain of mice was significantly up-regulated.And three forms ofα-synuclein protein expression significantly increased.Conclusion Injec-ting LPS into the bilateral substantia nigra for 5 consecutive days induces a more stable subacute PD mouse model.