一种快速易行的帕金森病小鼠炎症模型构建

A rapid and simple method for establishing an inflammatory model of Parkinson’s disease in mice

为构建快速简便的帕金森病(PD)小鼠炎症模型,将造模组分为7、14、21、30 d组,以旷场、爬杆和滚筒试验观察小鼠行为学变化;HPLC-ECD法测定多巴胺(DA)及代谢产物含量;免疫组织化学法观察TH阳性细胞数、NeuN神经元和Iba-1细胞数;Western blot检测TH、NeuN和NF-κB蛋白表达;ELISA法测定TNF-α、IL-1β含量。结果表明:造模组小鼠黑质致密部TH免疫阳性细胞数和纹状体DA含量均呈时间依赖性下降,同时,黑质和纹状体的TH蛋白表达也下调;而皮层和海马TH阳性细胞数和TH蛋白表达均与对照组无显著差异,皮层NeuN神经元数亦无明显变化,但海马区显著减少。此外, LPS使黑质Iba-1阳性细胞数、NF-κB p65蛋白表达和TNF-α、IL-1β含量均显著增加。这一研究提示此法能选择性引起小鼠黑质-纹状体DA能神经元损伤,且时间短、操作简便,可用于PD小鼠炎症模型的建立。

To establish a rapid and simple method for an inflammatory model of PD induced by LPS in mice. The mice were divided into the control group and the model groups(7, 14, 21, 30 d group), respectively. Open-field test, pole climb test and rotarod test were applied to detect the behavioral changes in mice. The contents of DA and its metabolites were measured by HPLC-ECD. The numbers of NeuN neurons, TH and Iba-1 positive cells were investigated by immunohistochemistry. Protein expressions of TH, NeuN and NF-κB were determined by Western blot. ELISA was used to survey the levels of TNF-α and IL-1β. The results showed that the numbers of TH immunoreactive cells of SNcp and DA contents of the Str in the mice induced by LPS were decreased in a time-dependent manner 30 days after LPS. At the same time, the expressions of TH protein in the SN and Str were also down-regulated. However, there was no significant difference in the numbers of TH immunoreactive cells and the expressions of TH protein in cortex and hippocampus of mice in LPS groups, compared with those in the control group. It was also found that the numbers of NeuN neurons in hippocampus were significantly reduced, while those of cortex had no significant change. In addition, LPS significantly increased the numbers of Iba-1 positive cells, expressions of NF-κB p65 protein, and levels of TNF-α and IL-1β in SN. The study showed that LPS, given according to the administration method applied in this experiment, can specifically cause damage to DA neurons in the substantia nigra-striatum of mice. The modeling approach is easy to operate and can be used to establish an inflammatory model of PD in mice.