脑血疏口服液灌胃对帕金森病模型大鼠黑质内神经炎症反应的影响

目的:研究脑血疏口服液对帕金森病(PD)模型大鼠黑质内神经炎症的抑制作用,以及对黑质多巴胺(DA)能神经元的保护作用。方法:应用立体定向注射技术,向大鼠脑右侧纹状体内注射6-羟多巴胺制备PD模型。将PD模型大鼠随机分为PD组、雷沙吉兰组和脑血疏口服液高、中、低剂量干预组,每组10只;另选10只正常大鼠作为对照组。采用免疫组织化学法检测各组大鼠黑质DA能神经元(酪氨酸羟化酶(TH)阳性)、小胶质细胞(CD11b阳性)、星形胶质细胞(GFAP阳性)形态变化。Western Blot法检测中脑黑质环氧化酶-2(COX-2)和肿瘤坏死因子-α(TNF-α)的变化。结果:大鼠行为学检测和黑质TH免疫组织化学染色显示,中剂量组的干预效果最好。免疫组织化学显示,与对照组相比,PD组的星形胶质细胞、小胶质细胞、TNF-α阳性细胞和COX-2阳性细胞均明显增加,TH阳性细胞数量明显减少,经中剂量的脑血疏口服液干预后,两种胶质细胞、TNF-α和COX-2阳性细胞数量均减少,TH阳性细胞增多。Western Blot显示,与PD组比较,中剂量组和雷沙吉兰组GFAP、CD11b、TNF-α和COX-2蛋白表达量显著减少(P0.05),而中剂量组与雷沙吉兰组比较上述蛋白表达均无显著差异(P0.05)。结论:脑血疏口服液可能通过抑制PD大鼠黑质胶质细胞的异常激活,减少炎性因子分泌而发挥神经保护作用。

The influence of Naoxueshu Oral Liquid on neuroinflammatory reaction of substantia nigra in rats with 6-OHDA-induced Parkinson's disease

Objective:To investigate the anti-inflammatory and neuroprotective effects of Naoxueshu Oral Liquid on 6-OHDA rat model of Parkinson's disease (PD).Methods:Male adult Sprague-Dawley rats were stereotactically injected with 6-hydroxydopamine (6-OHDA) into the right striatum.PD rats were randomly divided into PD group,Rasagiline group,low-,medium-and high-dose group.Ten rats were in each group.Ten adult normal rats were injected with saline instead of 6-OHDA as the control group.The morphological changes of dopaminergic neurons (tyrosine hydroxylase positive,TH-positive),astrocytes (GFAP-positive) and microglia (CD11 b-positive) in substantianigra of rats in each group were examined by immunohistochemical methods.The expression changes of tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2) were detected by Western Blot.Results:The results of behavioral testing and TH-immunoactive cells examination showed that the medium-dose Naoxueshu oral Liquid had the best therapeutic effects on PD rats (P＜0.05).Compared with the control group,the number of GFAP-,CD11b-,TNF-α-,COX-2-positive cells in substantianigra of the model group increased significantly,and the numbers of TH-positive cells was significantly decreased in PD model group.The numbers of astrocyte,microglia,TNF-α-and COX-2-positive cells were reduced,and the number of TH-positive cell increased in medium-dose group.The Western Blot showed that the expressions of GFAP,CD11 b,TNF-α and COX-2 in medium-dose group and Rasagiline group were significantly decreased compared with the PD group (P ＜ 0.05).There were no differences between the medium-dose group and Rasagiline group (P ＞ 0.05).Conclusion:Naoxueshu oral liquid might play a neuroprotective role by inhibiting abmormol activation of microglia and astrocytes,alleviating the inflammatory response in substantia nigra of PD rats.