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18～20 g,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:腹腔注射滗神经毒素,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,3 d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5 d重复注射,共5次;3 d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20 mg/kg及30 mg/kg.方法:各组小鼠实验前及最后一次注射神经毒素后3 d,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3 d,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.
揹景:帕金森病的主要病理改變在于中腦黑質的多巴胺神經元不可逆轉的變性減少,而氧化應激反應在帕金森病的髮病過程中起著十分重要的作用.3-硝基丙痠為線粒體複閤體-Ⅰ抑製劑,它可抑製氧化燐痠化作用從而抑製能量代謝,但德國Riepe教授髮現小劑量的3-硝基丙痠可以提高神經元對缺血缺氧的耐受性,它是否可以增彊多巴胺神經元對神經毒素的耐受性尚不得而知.目的:認識3-硝基丙痠多次預處理對帕金森病的預防及可能的機製.設計:對照觀察動物實驗.單位:華中科技大學同濟醫學院附屬協和醫院神經科.材料:實驗于2004-03/07在華中科技大學同濟醫學院附屬協和醫院神經科實驗室進行.C57BL小鼠48隻,體質量18～20 g,鼠齡二三箇月,雌雄不限.小鼠隨機分為6組,每組8隻:①空白對照組:不用任何藥物.②單次3-硝基丙痠組:僅腹腔註射3-硝基丙痠1次.③多次3-硝基丙痠組:每隔5d腹腔註射3-硝基丙痠,共5次.④神經毒素組:腹腔註射潷神經毒素,1次/d,共5次.⑤3-硝基丙痠單次預處理組:腹腔註射3-硝基丙痠1次,3 d後腹腔註射神經毒素,1次/d,共5次.⑥3-硝基丙痠多次預處理組:腹腔註射3-硝基丙痠,每隔5 d重複註射,共5次;3 d後再腹腔註射神經毒素,1次/d,共5次.3-硝基丙痠和神經毒素腹腔註射劑量分彆為20 mg/kg及30 mg/kg.方法:各組小鼠實驗前及最後一次註射神經毒素後3 d,分彆應用爬桿實驗及懸掛實驗對小鼠進行運動協調評分.各組小鼠在全部藥物註射完畢後3 d,迅速處死,測定中腦黑質丙二醛及還原型穀胱甘肽含量.主要觀察指標:①各組小鼠動作行為評分比較.②各組小鼠中腦黑質組織丙二醛含量的比較.③各組小鼠中腦黑質還原型穀胱甘肽含量的比較.結果:48隻小鼠均進入結果分析.①經腹腔註射神經毒素後,小鼠爬桿實驗,懸掛實驗較對照組評分降低(P＜0.01);經過3-硝基丙痠單/多次預處理後,其評分明顯上升,差異均有顯著性意義(P＜0.05,P＜0.01);多次預處理組與單次預處理組間差異也有顯著性意義(P＜0.05).②腹腔註射神經毒素後丙二醛含量較對照組明顯升高,差異有極顯著性意義(P＜0.01);3-硝基丙痠單次預處理後,丙二醛含量明顯下降,與神經毒素組比較,差異有顯著性意義(P＜0.05);3-硝基丙痠多次預處理後,丙二醛含量下降更為明顯,與神經毒素組比較,差異有極顯著性意義(P＜0.01);多次預處理組與單次預處理組比較,差異也有顯著性意義(P＜0.05).③與空白對照組比較,小鼠在單次腹腔註射3-硝基丙痠後,還原型穀胱甘肽含量沒有明顯變化;多次腹腔註射3-硝基丙痠後,還原型穀胱甘肽水平明顯升高(P＜0.05).在神經毒素組,還原型穀胱甘肽含量較空白對照組明顯下降(P＜0.01);3-硝基丙痠單次預處理後,還原型穀胱甘肽含量與神經毒素組相比無明顯變化(P＞0.05);3-硝基丙痠多次預處理後,還原型穀胱甘肽含量明顯升高,差異有顯著性意義(P＜0.05),多次預處理組與單次預處理組比較,差異有顯著性意義(P＜0.05).結論:3-硝基丙痠多次預處理可通過減少丙二醛生成,激髮還原型穀胱甘肽閤成,保護多巴胺神經元,達到預防帕金森病的作用.
배경:파금삼병적주요병리개변재우중뇌흑질적다파알신경원불가역전적변성감소,이양화응격반응재파금삼병적발병과정중기착십분중요적작용.3-초기병산위선립체복합체-Ⅰ억제제,타가억제양화린산화작용종이억제능량대사,단덕국Riepe교수발현소제량적3-초기병산가이제고신경원대결혈결양적내수성,타시부가이증강다파알신경원대신경독소적내수성상불득이지.목적:인식3-초기병산다차예처리대파금삼병적예방급가능적궤제.설계:대조관찰동물실험.단위:화중과기대학동제의학원부속협화의원신경과.재료:실험우2004-03/07재화중과기대학동제의학원부속협화의원신경과실험실진행.C57BL소서48지,체질량18～20 g,서령이삼개월,자웅불한.소서수궤분위6조,매조8지:①공백대조조:불용임하약물.②단차3-초기병산조:부복강주사3-초기병산1차.③다차3-초기병산조:매격5d복강주사3-초기병산,공5차.④신경독소조:복강주사필신경독소,1차/d,공5차.⑤3-초기병산단차예처리조:복강주사3-초기병산1차,3 d후복강주사신경독소,1차/d,공5차.⑥3-초기병산다차예처리조:복강주사3-초기병산,매격5 d중복주사,공5차;3 d후재복강주사신경독소,1차/d,공5차.3-초기병산화신경독소복강주사제량분별위20 mg/kg급30 mg/kg.방법:각조소서실험전급최후일차주사신경독소후3 d,분별응용파간실험급현괘실험대소서진행운동협조평분.각조소서재전부약물주사완필후3 d,신속처사,측정중뇌흑질병이철급환원형곡광감태함량.주요관찰지표:①각조소서동작행위평분비교.②각조소서중뇌흑질조직병이철함량적비교.③각조소서중뇌흑질환원형곡광감태함량적비교.결과:48지소서균진입결과분석.①경복강주사신경독소후,소서파간실험,현괘실험교대조조평분강저(P＜0.01);경과3-초기병산단/다차예처리후,기평분명현상승,차이균유현저성의의(P＜0.05,P＜0.01);다차예처리조여단차예처리조간차이야유현저성의의(P＜0.05).②복강주사신경독소후병이철함량교대조조명현승고,차이유겁현저성의의(P＜0.01);3-초기병산단차예처리후,병이철함량명현하강,여신경독소조비교,차이유현저성의의(P＜0.05);3-초기병산다차예처리후,병이철함량하강경위명현,여신경독소조비교,차이유겁현저성의의(P＜0.01);다차예처리조여단차예처리조비교,차이야유현저성의의(P＜0.05).③여공백대조조비교,소서재단차복강주사3-초기병산후,환원형곡광감태함량몰유명현변화;다차복강주사3-초기병산후,환원형곡광감태수평명현승고(P＜0.05).재신경독소조,환원형곡광감태함량교공백대조조명현하강(P＜0.01);3-초기병산단차예처리후,환원형곡광감태함량여신경독소조상비무명현변화(P＞0.05);3-초기병산다차예처리후,환원형곡광감태함량명현승고,차이유현저성의의(P＜0.05),다차예처리조여단차예처리조비교,차이유현저성의의(P＜0.05).결론:3-초기병산다차예처리가통과감소병이철생성,격발환원형곡광감태합성,보호다파알신경원,체도예방파금삼병적작용.
BACKGROUND: Mainly pathological 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-nitropropionie 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 Lab oratory, Union Hospital affiliated to Tongji Medical College, Huazhong U niversity 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 con trol group: Mice were not medicated. ② 3-NP single administration group: Mice were intraperitoneally injected with 3-NP once. ③ 3-NP repetitively administrations group: Mice were intraperitoneally injected with 3-NP every 5 days for 5 times in total. ④ Neurotoxin group: Mice were intraperitoneally injected with neurotoxin once every day for 5 times in total. ⑤ 3-NP single preconditioning group: Mice were intraperitoneal ly injected with 3-NP once, and 3 days later, they were intraperitoneally injected with neurotoxin once every day for 5 times in total. ⑥ 3-NP repetitively preconditionings 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. METHODS: 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 rapid ly to measure the contents of malondialdehyde (MDA) and reduced glu tathione (GSH) in the substantia nigra of midbrain. MAIN OUTCOME MEASURES: ① Motor and behavior scores; ② con tent of MDA; ③ content of GSH.~ULTS: All 48 mice were involved in the final analysis. ① Scores 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 differencebetween 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.
