葛根素对帕金森病大鼠黑质组织HO-1,NQO1表达的影响

目的:研究葛根素对6-羟多巴胺(6-OHDA)诱导帕金森大鼠黑质组织血红素加氧酶-1(HO-1),醌氧化还原酶(NQO1)表达的影响.方法:建立帕金森SD大鼠模型,随机分成5组:模型组、美多巴阳性组(40 mg·kg-1)及葛根素低、中、高剂量组(20,40,80 mg·kg-1).持续灌胃给药30 d.检测黑质组织中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量.HE染色观察黑质神经细胞形态学的变化.RT-PCR法检测黑质组织HO-1mRNA表达.Western blot法检测NQO1蛋白表达.结果:与模型组比较,葛根素显著地增加帕金森病大鼠黑质GSH-Px活性(368.29±37.86),(501.74±43.62),(581.43 ±50.97) U·mg-1和SOD活性(119.36±10.24),(179.85±13.04),(206.94±14.37)NU·mg-1,同时降低MDA含量(10.47±1.15),(7.02±0.94),(5.38±0.82)nmol.mg-1(P＜0.01).缓解6-OHDA所致黑质神经细胞损伤.有效下调黑质HO-1 mRNA水平42.1％,34.2％,24.7％ (P ＜0.01),以及明显增加NQO1蛋白表达24.3％,30.9％,47.6％(P＜0.01).结论:葛根素有效逆转6-OHDA致PD大鼠黑质神经细胞损伤,其机制可能与其抑制氧化应激途径有关.

Effect of Puerarin on Expressions of HO-1,NQO1 in Substantia Nigra Tissue of Parkinson's Rats

Objective:To investigate the effect of puerarin on the expressions of heme oxygenase-1 (HO-1), quinone oxidoreductase-1 (NQO-1) in substantia nigra tissue of Parkinson's rat model induced by 6-hydroxydopamine (6-OHDA). Method: Parkinson's rat model was established, the rats were randomly divided into 5 groups: model group, Madopar group (40 mg·kg^-1), low-, medium-and high-dosage groups of puerarin (20, 40, 80 mg·kg^-1). The drugs were intragastrically perfused to rats daily for 30 consecutive days. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) as well as the contents of malonaldehyde (MDA) in substantia nigra tissue were tested by biochemical method. The morphologic changes in nerve cells of substantia nigra were observed by HE staining. The expression of HO-1 mRNA was tested by RT-PCR assay. And the NQO-1 protein expression was tested by in situ hybridization analysis. Result: Compared to model group, puerarin effectively increased the activities of GSH-Px and SOD in substantia nigra tissue of parkinson's rats, while the MDA content was reduced (P<0.01). And alleviated the nerve cell damages in substantia nigra induced by 6-OHDA. And the level of HO-1 mRNA in substantia nigra was down-regulated (P<0.01). The expression of NQO-1 protein was notably up-regulated. Conclusion: The findings indicate that puerarin effectively reverse the neuronal cells injure in substantia nigra of Parkinson's disease(PD) rats induced by 6-OHDA, and its underlying mechanism may be linked to suppression of oxidative stress pathway.