摘要
背景:帕金森病的主要病理改变在于中脑黑质的多巴胺神经元不可逆转的变性减少,而氧化应激反应在帕金森病的发病过程中起着十分重要的作用。3-硝基丙酸为线粒体复合体-I抑制剂,它可抑制氧化磷酸化作用从而抑制能量代谢,但德国Riepe教授发现小剂量的3-硝基丙酸可以提高神经元对缺血缺氧的耐受性,它是否可以增强多巴胺神经元对神经毒素的耐受性尚不得而知。目的:认识3-硝基丙酸多次预处理对帕金森病的预防及可能的机制。设计:对照观察动物实验。单位:华中科技大学同济医学院附属协和医院神经科。材料:实验于2004-03/07在华中科技大学同济医学院附属协和医院神经科实验室进行。C57BL小鼠48只,体质量18～20g,鼠龄二三个月,雌雄不限。小鼠随机分为6组,每组8只:①空白对照组:不用任何药物。②单次3-硝基丙酸组:仅腹腔注射3-硝基丙酸1次。③多次3-硝基丙酸组:每隔5d腹腔注射3-硝基丙酸,共5次。④神经毒素组:腹腔注射神经毒素,1次/d,共5次。⑤3-硝基丙酸单次预处理组:腹腔注射3-硝基丙酸1次,3d后腹腔注射神经毒素,1次/d,共5次。⑥3-硝基丙酸多次预处理组:腹腔注射3-硝基丙酸,每隔5d重复注射,共5次;3d后再腹腔注射神经毒素,1次/d,共5次。3-硝基丙酸和神经毒素腹腔注射剂量分别为20mg/kg及30mg/kg。方法:各组小鼠实验前及最后一次注射神经毒素后3d,分别应用爬杆实验及悬挂实验对小鼠进行运动协调评分。各组小鼠在全部药物注射完毕后3d,迅速处死,测定中脑黑质丙二醛及还原型谷胱甘肽含量。主要观察指标:①各组小鼠动作行为评分比较。②各组小鼠中脑黑质组织丙二醛含量的比较。③各组小鼠中脑黑质还原型谷胱甘肽含量的比较。结果:48只小鼠均进入结果分析。①经腹腔注射神经毒素后,小鼠爬杆实验,悬挂实验较对照组评分降低(P<0.01);经过3-硝基丙酸单/多次预处理后,其评分明显上升,差异均有显著性意义(P<0.05,P<0.01);多次预处理组与单次预处理组间差异也有显著性意义(P<0.05)。②腹腔注射神经毒素后丙二醛含量较对照组明显升高,差异有极显著性意义(P<0.01);3-硝基丙酸单次预处理后,丙二醛含量明显下降,与神经毒素组比较,差异有显著性意义(P<0.05);3-硝基丙酸多次预处理后,丙二醛含量下降更为明显,与神经毒素组比较,差异有极显著性意义(P<0.01);多次预处理组与单次预处理组比较,差异也有显著性意义(P<0.05)。③与空白对照组比较,小鼠在单次腹腔注射3-硝基丙酸后,还原型谷胱甘肽含量没有明显变化;多次腹腔注射3-硝基丙酸后,还原型谷胱甘肽水平明显升高(P<0.05)。在神经毒素组,还原型谷胱甘肽含量较空白对照组明显下降(P<0.01);3-硝基丙酸单次预处理后,还原型谷胱甘肽含量与神经毒素组相比无明显变化(P>0.05);3-硝基丙酸多次预处理后,还原型谷胱甘肽含量明显升高,差异有显著性意义(P<0.05),多次预处理组与单次预处理组比较,差异有显著性意义(P<0.05)。结论:3-硝基丙酸多次预处理可通过减少丙二醛生成,激发还原型谷胱甘肽合成,保护多巴胺神经元,达到预防帕金森病的作用。
BACKGROUND： Mainly patholog/cal changes of Parkinson disease （PD） are related to irreversible degeneration and reduction of dopamine neurons of substantia nigra in midbrain; however, oxidative stress reaction plays an important role in onset of PD. 3-nitropropionic acid （3-NP） is an inhibitor of mitochondria compound Ⅰ, and it can inhibit oxidative phosphorylation so as to restrain energy metabolism. However, professor Riepe from Germany found that small dose of 3-NP can increase the tolerance of neurons to ischemic hypoxia. It is unclear whether it can also strengthen the tolerance of dopamine neurons to neurotoxin. OBJECTIVE： To investigate the possible mechanism and prevention of repetitively preconditioning 3-NP for treating PD. DESIGN： Controlled observational animal study.SETTING： Department of Neurology, Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： The experiment was carried out at the Neurological Laboratory, Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology from March to July 2004. A total of 48 C57BL mice, weighing 18-20 g, aged 2-3 months, of both genders, were randomly divided into 6 groups with 8 in each group.① Blank control group： Mice were not medicated. ② 3-NP single administration group： Mice were intraperitoneally injected with 3-NP once. ③3-NP repetitively administrations group： Mice were intraperitoneally injected with 3-NP every 5 days for 5 times in total. ④Neurotoxin group： Mice were intraperitoneally injected with neurotoxin once every day for 5. times in total. ⑤3-NP single preconditioning group： Mice were intraperitoneally injected with 3-NP once, and 3 days later, they were intraperitoneally injected with neurotoxin once every day for 5 times in total. ⑥3-NP repetitively precoriditionings group： Mice were intraperitoneally injected with 3-NP and repetitively every 5 days for 5 times in total; 3 days later, mice were intraperitoneally injected with neurotoxin once every day for 5 times in total. Dosages of 3-NP and neurotoxin were 20 mg/kg and 30 mg/kg, respectively. METHODSt Motor coordination of mice was scored with pole test and traction test before experiment and at 3 days after the last injection of neurotoxin. Three days after complete injection, mice were sacrificed rapidly to measure the contents of malondialdehyde （MDA） and reduced glutathione （GSH） in the substantia nigra of midbrain. MAIN OUTCOME MEASURES： ①Motor and behavior scores; ②content of MDA; ③content of GSH.RESULTS： All 48 mice were involved in the final analysis. ①Scores of pole test and traction test were decreased in neurotoxin group as compared with those in control group （P 〈 0.01）; but the scores were increased after 3-NP single/repetitively preconditionings, and there were significant difference （P 〈 0.05, P 〈 0.01）. Meanwhile, there was also significant difference between 3-NP repetitively preconditionings group and 3-NP single preconditioning group （P 〈 0.05）.② Content of MDA was increased in neurotoxin group as compared with that in control group, and there was significant difference （P 〈 0.01）; content of MDA was decreased after 3-NP single preconditioning as compared with that in neurotoxin group, and there was significant difference （P 〈 0.05）; content of MDA was remarkably decreased after 3-NP repetitively preconditionings as compared with that in neurotoxin group, and there was greatly significant difference （P 〈 0.01）; meanwhile, there was also significant difference between 3-NP repetitively preconditionings group and 3-NP single preconditioning group （P 〈 0.05）. ③As compared with that in blank control group, content GSH in 3-NP single administration group was not changed; content of GSH in 3-NP repetitively administrations group was increased （P 〈 0.05）; content of GSH in neurotoxin group was decreased as compared with that in blank control group （P 〈 0.01）; content of GSH in 3-NP single preconditioning group was not changed as compared with that in neurotoxin group （P 〉 0.05）; content of GSH was increased after 3-NP repetitively preconditionings, and there was significant difference （P 〈 0.05）; meanwhile, there was significant difference between 3-NP repetitively preconditionings group and 3-NP single preconditioning group （P 〈 0.05）. CONCLUSION： 3-NP repetitively preconditionings can activate synthesis of GSH, protect dopamine neurons through decreasing production of MDA.
出处
《中国临床康复》
CSCD
北大核心
2006年第34期180-183,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30170334)~~
