6-羟基多巴胺帕金森病模型大鼠氧化应激反应

Oxidative stress reaction of rats with 6-hydroxydopamine Parkinson disease

目的:观察6-羟基多巴胺帕金森病大鼠模型的氧化应激反应及其可能的机制。方法:实验于2005-06/12在上海中医药大学科研实验中心完成。实验分组:Wistar雄性大鼠18只,随机分为正常对照组、假手术组、模型组,每组6只。实验干预:采用6-羟基多巴胺(溶于0.2g/L抗坏血酸的生理盐水中)注射于脑右侧黑质,10d后以腹腔注射阿朴吗啡0.5mg/kg诱发大鼠向一侧旋转,记录开始旋转至30min内的旋转圈数,以平均每分钟旋转圈数超过7次者为合格的帕金森病模型。假手术组6只大鼠只注射含0.2g/L抗坏血酸的等量生理盐水,其余条件与造模组手术相同。正常对照组6只大鼠只进行大鼠固定,不进行任何处理。在旋转诱发实验完成后,动物每天自由饮水与进食,共45d。实验评估:应用化学比色法测定大鼠中脑黑质纹状体部位活性氧、丙二醛、谷胱甘肽含量,谷胱甘肽过氧化物酶、超氧化物歧化酶的活性。结果:18只大鼠均进入结果分析。①活性氧含量:模型组明显高于正常对照组和假手术组[(451.13±73.42),(135.62±53.46),(161.15±61.16)U/mg,P<0.01]。②谷胱甘肽含量:模型组明显低于正常对照组和假手术组[(4.26±0.75),(7.03±1.39),(7.03±1.42)mg/g,P<0.05]。③谷胱甘肽过氧化物酶活性:模型组明显低于正常对照组和假手术组[(3.05±0.38),(3.96±0.39),(3.77±0.38)NU/g,P<0.01]。④超氧化物歧化酶活性:模型组明显低于正常对照组和假手术组[(137.74±4.65),(190.77±8.47),(199.73±8.23)NU/mg,P<0.01]。⑤丙二醛含量:模型组明显高于正常对照组和假手术组[(10.90±2.17),(4.18±2.88),(4.52±2.45)μmol/g,P<0.01]。结论:帕金森病模型大鼠的氧化反应被激活,处于氧化应激状态,氧化应激反应增强可能是帕金森病的发病机制之一。

AIM： To investigate the oxidative stress reaction of rats with 6-hydroxydopamine （6-OHDA） Parkinson disease （PD） and its potential mechanism. METHODS： The experiment was completed in the Scientific Research Laboratory Center of Shanghai University of Traditional Chinese Medicine from June to December in 2005. Eighteen Wistar male rats were randomized into three groups： normal control group, sham-operated group and PD model group. 6-OHDA （dissolved in normal saline containing 0.2 g/L antiscorbic acid） was injected into the right substantial nigra （RSN）. Ten days later, 0.5 mg/kg apomorphine （APO） was injected intraperitoneally to induce lateral rotation of the rats. Finally the rotation circles from the starting point to 30 minutes were recorded, and those with more than 7 circles per minute on average were regarded as qualified PD model. Except for injection of same amount of normal saline containing 0.2 g/L antiscorbic acid, other managements of the sham-operated group were exactly same as the model group. The rats in normal control group were only done immobilization. Following the APO-induced rotation experiment, the rats were given water and food intake freely for 45 days. Then the rats in each group were decapitated. And the contents of active oxygen in nigrostriatal area, malondialdehyde （MDA）, glutathione （GSH）,. GSH peroxidase （GSH-Px）, and superoxidedismutase （SOD） were measured with chemical chromatometry. RESULTS： All the 18 rats were involved in the result analysis. （1）The active oxygen in the PD model group was significantly elevated compared with these of normal control group and sham-operated group [（451.13±73.42）, （135.62± 53.46）, （161.15±61.16） U/mg, P 〈 0.01].（2）The GSH activity in the PD model group was significantly reduced compared with these of normal control group and sham-operated group [（4.26±0.75）, （7.03±1.39）, （7.03±1.42） mg/g, P 〈 0.05].（3） The GSH-Px activity in the PD model group was significantly reduced compared with these of normal control group and sham-operated group [（3.05±0.38）, （3.96±0.39）, （3.77±0.38） NU/g, P 〈 0.01].（4）The SOD activity in the PD model group was significantly lower than those of normal control group and sham-operated group [（137.74±4.65）,（190.77±8.47）, （199.73±8.23） NU/mg, P 〈 0.01].（5）The MDA content in the PD model group was significantly higher than these of normal control group and sham-operated group [（10.90±2.17）, （4.18±2.88）, （4.52±2.45） μmol/g, P 〈 0.01]. CONCLUSION： The many signs of increased oxidative stress reaction found in rat model of PD are suggestive of their contribution to the etiology of PD.