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目的:研究肾茶总黄酮对6-羟基多巴胺(6-OHDA)诱致帕金森病(Parkinson’s disease,PD)大鼠模型及细胞模型的保护作用。方法:Wistar雄性大鼠80只随机分为造模组和正常组,造模组70只,正常组10只,除正常组外,采用定位注射6-OHDA(8μg溶于4μL含质量分数为0.2%抗坏血酸的生理盐水中)损毁大鼠单侧黑质多巴胺(DA)神经元的方法建立PD大鼠模型,将造模成功的大鼠随机分成5组,即模型组、肾茶总黄酮高、中、低剂量组(45,22,11 mg·kg-1·d-1)、美多芭组(7.8 mg·kg-1·d-1),给药组ig给予相应药物,给药14 d,给药结束后,进行大鼠行为学检测并处死,采用ELISA法测定大鼠脑组织中丙二醛(MDA),过氧化氢酶(CAT),谷胱甘肽过氧化物酶(GSH-Px),超氧化物歧化酶(SOD),DA和高香草酸(HVA)多项指标的水平。以神经母细胞瘤细胞(SH-SY5Y细胞)为研究对象,实验分为空白组、模型组、肾茶总黄酮2,1,0.5,0.25 g·L-1组,共6组,除空白组外,各组加入30μmol·L-1的6-OHDA,复制PD细胞模型,肾茶总黄酮各剂量组均经过不同浓度肾茶总黄酮预处理,继续培养20 h后,对细胞存活率和细胞形态进行观察比较。结果:与正常组比较,模型组PD大鼠的旋转次数增加,脑组织CAT,GSH-Px,SOD和HVA水平明显降低,MDA水平明显升高,均具有统计学差异(P<0.01);与模型组比较,肾茶总黄酮高剂量可以明显减少PD大鼠旋转次数,肾茶总黄酮高、中、低剂量组均可不同程度的提高脑组织CAT,GSH-Px,SOD和HVA水平,降低MDA水平(P<0.05,P<0.01),但是在改善神经行为和提高DA水平方面作用效果不及美多芭明显。与空白组比较,模型组6-OHDA引起的细胞损伤较明显(P<0.05);与模型组比较,肾茶总黄酮(2,1 g·L-1)组预处理组可以明显减轻6-OHDA引起的细胞损伤(P<0.05,P<0.01)。结论:肾茶总黄酮对6-OHDA诱导的PD大鼠模型和细胞模型具有明显的保护作用,其可能的作用机制与减少抗氧化应激引起的细胞损伤相关。
OBJECTIVE: To study the protective effect of total flavones from Shenchao on the rat model and cell model of Parkinson’s disease (PD) induced by 6-hydroxydopamine (6-OHDA). Methods: Eighty Wistar male rats were randomly divided into model group and normal group. There were 70 model rats and 10 normal rats. Normal rats were injected with 6-OHDA (8μg dissolved in 4μL with a mass fraction of 0.2 % Ascorbic acid in normal saline) to destroy unilateral substantia nigra dopamine (DA) neurons in rats. To establish a rat model of PD, the rats with successful modeling were randomly divided into 5 groups: model group, Middle and low dose groups (45, 22, and 11 mg · kg-1 · d-1), and metoprolol group (7.8 mg · kg-1 · d-1) d. After the administration, behavioral tests were performed and sacrificed. The contents of malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GSH) -Px), superoxide dismutase (SOD), DA and high vanillic acid (HVA) a number of indicators. The neuroblastoma cells (SH-SY5Y cells) were divided into 6 groups: blank group, model group and total flavones of Shencha (2,1,0.5,0.25 g · L -1) In addition, 30μmol·L-1 6-OHDA was added into each group to replicate the PD cell model. The total flavonoids of Shencha were pretreated with different concentrations of total flavonoids of Shencha, and after 20 hours of continuous culture, the cell viability and cells Morphological observation and comparison. Results: Compared with the normal group, the number of rotations, the levels of CAT, GSH-Px, SOD and HVA in brain tissue of rats in model group were significantly decreased, and the levels of MDA were significantly increased (all P <0.01) Compared with the model group, the high dose of total flavonoids of Shencha could obviously reduce the number of rotations of PD rats. The levels of CAT, GSH-Px, SOD and HVA in brain tissues of rats in high, medium and low dose of total flavonoids of Shencha can all reduce to some extent MDA level (P <0.05, P <0.01), but the effect was not as obvious as that of methdopa in improving neurological behavior and increasing DA level. Compared with the blank group, the cell injury caused by 6-OHDA in the model group was more obvious (P <0.05). Compared with the model group, the pretreatment with 2,1 g · L-1 group could significantly reduce the 6- OHDA induced cell injury (P <0.05, P <0.01). Conclusion: The total Flavone of Shencha has significant protective effect on 6-OHDA-induced PD rat model and cell model, and its possible mechanism is related to the reduction of cell injury caused by anti-oxidative stress.