帕病2号方对帕金森病模型大鼠中脑黑质Nrf2，HO-1表达的影响

Research on Expressions of Nrf2 and HO-1 in Substantia Nigra of Rats with Parkinson Disease Treated with Pabing 2 Formula

摘要:

目的：观察帕病2号方对6-羟基多巴胺（6-OHDA）所致帕金森病（PD）大鼠中脑黑质组织内核因子E2相关性因子2（Nrf2）以及下游靶基因血红加氧酶1（HO-1）的表达。方法：采用6-OHDA左侧纹状体两点注射法建立PD大鼠模型，经阿卟吗啡（APO）诱导表现为恒定右侧旋转且旋转圈数大于210 r·30 min^-1视为成功PD大鼠模型。24只造模成功大鼠随机分为模型组，帕病2号方低、中、高剂量组（8.0，16.0，32.0 g·kg^-1），同时设立正常组，假手术组。正常组，假手术组，模型组给予等容积蒸馏水，连续灌胃给药4周。Western blot 法检测中脑黑质Nrf2细胞核蛋白及HO-1蛋白表达。RT-PCR法检测中脑黑质Nrf2，HO-1mRNA表达。结果：与假手术组比较，模型组Nrf2细胞核蛋白，HO-1蛋白表达上调（P<0.01），Nrf2，HO-1mRNA水平上调（P<0.01）；与模型组比较，帕病2号方低、中、高剂量组大鼠中脑黑质组织Nrf2细胞核蛋白，HO-1蛋白表达明显上调（P<0.01），Nrf2，HO-1mRNA水平上调（P<0.01），帕病2号方高剂量组与中、低剂量组比较有明显差异（P<0.01）。结论：6-OHDA造模后可以激活Nrf2/HO-1信号通路；帕病2号方治疗后进一步提高Nrf2细胞核蛋白和Nrf2mRNA表达，进而上调下游靶基因HO-1蛋白及HO-1mRNA的表达，且帕病2号方高剂量组作用更为明显。

Abstract:

Objective: To investigate the effect of Pabing 2 formula on the expression of nuclear factor E2-related factor 2(Nrf2)and the hemeoxygenasel(HO-1) in substantia nigra tissue of Parkinson disease(PD) rats model induced by 6-hydroxydopamine(6-OHDA). Method: The rats were stereotaxically injected with 6-OHDA solution into the left striatum in two-site.Rats showed consistently right whirling and the number of rotation was more than 210 r·30 min^-1 induced by APO, then the rat was judged as PD model.Twenty-four modeling rats successfully were randomly divided into four groups:model group, Pabing 2 formula(8.0, 16.0, 32.0 g·kg^-1), at the same time, the normal group and sham group were established and given distilled water at the same volume, all with a course of 4 weeks. The nuclear Nrf2 protein and HO-1 protein levels of the substantia nigra tissue were detected by Western blot, and the mRNA levels of Nrf2 and HO-1 were determined with RT-PCR. Result: Compared with sham group, the nuclear Nrf2 protein and HO-1protein in substantia nigra tissue were significant increased(P<0.01), and the mRNA levels of Nrf2 and HO-1 were increased in model group(P<0.01). Compared with model group, the nuclear Nrf2 protein and HO-1 protein in substantia nigra tissue were significantly increased (P<0.01), and the mRNA levels of both Nrf2 and HO-1were also upregulated in Pabing 2 formula low dose group, media dose group and high dose group(P<0.01), Pabing 2 formula high dose group were significantly differences compared with the low dose group and media dose group (P<0.01). Conclusion: After 6-OHDA injection, the Nrf2/HO-1 signal transduction pathway is activated.Pabing 2 formula can enhance antioxidation ability.The underlying mechanism might be associated with activating Nrf2/HO-1 signal transduction pathway, promoting the nuclear Nrf2 protein and Nrf2 mRNA, then advancing the expression of antioxidant gene in the downstream such as HO-1 protein and HO-1 mRNA.The effects were more obvious in Pabing 2 formula high dose group.