背景:有实验指出酸性肽可能是通过抑制一氧化氮等毒性化合物的生成而提高阿尔茨海默病模型大鼠的学习记忆能力.目的:建立阿尔茨海默病动物模型,观察给予不同剂量浓度的酸性肽治疗后大鼠脑一氧化氮、一氧化氮合酶与乙酰胆碱酯酶含量的变化.设计:随机对照单一实验.单位:郑州大学基础医学院生物化学与分子生物学教研室.材料:实验于2003-02/07在郑州大学基础医学院生物化学与分子生物学教研室的第二研究室和实验动物房完成.选取雄性SD大鼠100只,用跳台实验除去反应迟钝的动物,共84只大鼠纳入实验,随机分为7组:正常对照组,模型组,生理盐水组,吡拉西坦治疗组,酸性肽15,30,60mg/kg治疗组,12只/组.酸性肽为本课题组从牛脑中分离出的一个新的小分子肽,由三个谷氨酸连成的三肽.方法:除正常对照组外,其余各组大鼠常规饲养1周后,均采用大鼠脑组织立体定位微量注射技术,脑海马注射5 μg鹅膏蕈氨酸,以损毁大鼠双侧迈纳特基底核建立阿尔茨海默病模型.正常对照组和模型组不给药,生理盐水组用生理盐水灌胃,吡拉西坦治疗组用0.3 g/kg吡拉西坦灌胃,酸性肽15,30,60mg/kg治疗组分别用15,30,60mg/kg酸性肽灌胃,连续20 d,1次/d,2mL/次.灌胃期满将大鼠麻醉后断头处死,立即在冰盘中开颅取脑制备组织匀浆.4℃下1 000 r/min离心10 min,取上清液用一氧化氮、一氧化氮合酶和乙酰胆碱酯酶检测试剂盒测定一氧化氮、一氧化氮合酶和乙酰胆碱酯酶的含量.主要观测指标:各组大鼠脑内一氧化氮、一氧化氮合酶和乙酰胆碱酯酶的含量.结果:纳入实验的84只大鼠全部进入结果分析.各组大鼠脑内一氧化氮、一氧化氮合酶和乙酰胆碱酯酶含量的比较:与模型组比较,酸性肽15,30,60mg/kg治疗组一氧化氮含量均明显降低[(1.95±0.20),(1.39±0.10),(1.25±0.07),(1.00±0.04)mmoL/kg,P＜0.05];一氧化氮合酶含量均明显降低[(4.53±0.18),(3.39±0.09),(3.10±0.06),(2.97±0.06)μmol/kg,P＜0.05];乙酰胆碱酯酶含量无明显变化[(0.67±0.12),(0.71±0.11),(0.72±0.08),(0.72±0.07)mmol/L,P＞0.05].结论:酸性肽能够显著降低阿尔茨海默病模型大鼠脑内一氧化氮、一氧化氮合酶的生成,而对乙酰胆碱酯酶的活性并无明显影响.提示酸性肽可能是通过抑制一氧化氮等毒性化合物的生成或毒性作用来提高阿尔茨海默病模型大鼠的学习记忆能力.
揹景:有實驗指齣痠性肽可能是通過抑製一氧化氮等毒性化閤物的生成而提高阿爾茨海默病模型大鼠的學習記憶能力.目的:建立阿爾茨海默病動物模型,觀察給予不同劑量濃度的痠性肽治療後大鼠腦一氧化氮、一氧化氮閤酶與乙酰膽堿酯酶含量的變化.設計:隨機對照單一實驗.單位:鄭州大學基礎醫學院生物化學與分子生物學教研室.材料:實驗于2003-02/07在鄭州大學基礎醫學院生物化學與分子生物學教研室的第二研究室和實驗動物房完成.選取雄性SD大鼠100隻,用跳檯實驗除去反應遲鈍的動物,共84隻大鼠納入實驗,隨機分為7組:正常對照組,模型組,生理鹽水組,吡拉西坦治療組,痠性肽15,30,60mg/kg治療組,12隻/組.痠性肽為本課題組從牛腦中分離齣的一箇新的小分子肽,由三箇穀氨痠連成的三肽.方法:除正常對照組外,其餘各組大鼠常規飼養1週後,均採用大鼠腦組織立體定位微量註射技術,腦海馬註射5 μg鵝膏蕈氨痠,以損燬大鼠雙側邁納特基底覈建立阿爾茨海默病模型.正常對照組和模型組不給藥,生理鹽水組用生理鹽水灌胃,吡拉西坦治療組用0.3 g/kg吡拉西坦灌胃,痠性肽15,30,60mg/kg治療組分彆用15,30,60mg/kg痠性肽灌胃,連續20 d,1次/d,2mL/次.灌胃期滿將大鼠痳醉後斷頭處死,立即在冰盤中開顱取腦製備組織勻漿.4℃下1 000 r/min離心10 min,取上清液用一氧化氮、一氧化氮閤酶和乙酰膽堿酯酶檢測試劑盒測定一氧化氮、一氧化氮閤酶和乙酰膽堿酯酶的含量.主要觀測指標:各組大鼠腦內一氧化氮、一氧化氮閤酶和乙酰膽堿酯酶的含量.結果:納入實驗的84隻大鼠全部進入結果分析.各組大鼠腦內一氧化氮、一氧化氮閤酶和乙酰膽堿酯酶含量的比較:與模型組比較,痠性肽15,30,60mg/kg治療組一氧化氮含量均明顯降低[(1.95±0.20),(1.39±0.10),(1.25±0.07),(1.00±0.04)mmoL/kg,P＜0.05];一氧化氮閤酶含量均明顯降低[(4.53±0.18),(3.39±0.09),(3.10±0.06),(2.97±0.06)μmol/kg,P＜0.05];乙酰膽堿酯酶含量無明顯變化[(0.67±0.12),(0.71±0.11),(0.72±0.08),(0.72±0.07)mmol/L,P＞0.05].結論:痠性肽能夠顯著降低阿爾茨海默病模型大鼠腦內一氧化氮、一氧化氮閤酶的生成,而對乙酰膽堿酯酶的活性併無明顯影響.提示痠性肽可能是通過抑製一氧化氮等毒性化閤物的生成或毒性作用來提高阿爾茨海默病模型大鼠的學習記憶能力.
배경:유실험지출산성태가능시통과억제일양화담등독성화합물적생성이제고아이자해묵병모형대서적학습기억능력.목적:건립아이자해묵병동물모형,관찰급여불동제량농도적산성태치료후대서뇌일양화담、일양화담합매여을선담감지매함량적변화.설계:수궤대조단일실험.단위:정주대학기출의학원생물화학여분자생물학교연실.재료:실험우2003-02/07재정주대학기출의학원생물화학여분자생물학교연실적제이연구실화실험동물방완성.선취웅성SD대서100지,용도태실험제거반응지둔적동물,공84지대서납입실험,수궤분위7조:정상대조조,모형조,생리염수조,필랍서탄치료조,산성태15,30,60mg/kg치료조,12지/조.산성태위본과제조종우뇌중분리출적일개신적소분자태,유삼개곡안산련성적삼태.방법:제정상대조조외,기여각조대서상규사양1주후,균채용대서뇌조직입체정위미량주사기술,뇌해마주사5 μg아고심안산,이손훼대서쌍측매납특기저핵건립아이자해묵병모형.정상대조조화모형조불급약,생리염수조용생리염수관위,필랍서탄치료조용0.3 g/kg필랍서탄관위,산성태15,30,60mg/kg치료조분별용15,30,60mg/kg산성태관위,련속20 d,1차/d,2mL/차.관위기만장대서마취후단두처사,립즉재빙반중개로취뇌제비조직균장.4℃하1 000 r/min리심10 min,취상청액용일양화담、일양화담합매화을선담감지매검측시제합측정일양화담、일양화담합매화을선담감지매적함량.주요관측지표:각조대서뇌내일양화담、일양화담합매화을선담감지매적함량.결과:납입실험적84지대서전부진입결과분석.각조대서뇌내일양화담、일양화담합매화을선담감지매함량적비교:여모형조비교,산성태15,30,60mg/kg치료조일양화담함량균명현강저[(1.95±0.20),(1.39±0.10),(1.25±0.07),(1.00±0.04)mmoL/kg,P＜0.05];일양화담합매함량균명현강저[(4.53±0.18),(3.39±0.09),(3.10±0.06),(2.97±0.06)μmol/kg,P＜0.05];을선담감지매함량무명현변화[(0.67±0.12),(0.71±0.11),(0.72±0.08),(0.72±0.07)mmol/L,P＞0.05].결론:산성태능구현저강저아이자해묵병모형대서뇌내일양화담、일양화담합매적생성,이대을선담감지매적활성병무명현영향.제시산성태가능시통과억제일양화담등독성화합물적생성혹독성작용래제고아이자해묵병모형대서적학습기억능력.
BACKGROUND: It is pointed in some experiment that acidic peptide improves learning and memory of model rat with Alzheimer disease (AD) by inhibiting the synthesis of toxic compounds of nitric oxide (NO).OBJECTIVE: Animal model with Alzheimer disease was established to observe the changes in the levels of NO, nitric oxide synthase (NOS) and acetylcholinesterase (AChE) treated with acidic peptide of various dose concentration.DESIGN: Randomized control and single experiment.SETTING: Teaching-Research Room of Biochemistry and Molecular Biology of Basic Medical College of Zhengzhou University.MATERIALS: The experiment was performed in 2nd Research Room and Experimental Animal Room of Teaching-Research Room of Biochemistry and Molecular Biology of Basic Medical College of Zhengzhou University.Totally 100 SD male rats were selected and some of them were excluded due to retarded response in step down test. Totally 84 rats were included in the experiment and randomized into 7 groups, named normal control,model group, physiological saline group (PS group), Piracetam group, acidic peptide groups of 15, 30 and 60 mg/kg, 12 rats in each group. Acidic peptide was a new small molecular peptide separated from bovine brain and is tripeptide composed of three glutamic acids.METHODS: Except normal control, in the rest groups, after 1 week routine breeding, cerebral stereotactic microinjection was used to inject 5 μg ibotenic acid in hippocampus of rats to destroy bilateral Meynert's nucleus basalis to establish AD model. In normal control and model group, no medication was applied. In PS group, physiological saline was used for gastric perfusion. In piracetam group, piracetam of 0.3 g/kg was used for gastric perfusion and in acidic peptide groupsof 15, 30 and 60 mg/kg,acidic peptide of 15, 30 and 60 mg/kg was applied for gastric perfusion successively, continuously for 20 days, once per day, 2 mL/time. On the expiration of gastric perfusion, the rats were sacrificed after anesthetized and the brain was collected on ice plate to prepare tissue homogenate. After centrifugated at 1 000 r/minute, 4℃ for 10 minutes, the supernatant was collected to assay the levels of NO, NOS and AChE with NO, NOS and AChE kits successively.MAIN OUTCOME MEASURES: Levels of NO, NOS and AChE in brain of rat in each groupRESULTS: Totally 84 rats were employed in the experiment and all entered result analysis. Comparison of levels of NO, NOS and AChE in rat brain of each group: compared with model group, NO levels in acidic peptide groups of 15, 30 and 60 mg/kg were reduced remarkably[(1.95±0.20), (1.39±0.10), (1.25±0.07), (1.00±0.04) mmoL/kg, P ＜ 0.05],NOS levels were reduced remarkably [(4.53±0.18), (3.39±0.09), (3.10±0.06),(2.97±0.06) μmol/kg, P ＜ 0.05] and AChE did not change remarkably[(0.67±0.12), (0.71±0.11), (0.72±0.08), (0.72±0.07) mmol/L, P ＞ 0.05].CONCLUSION: Acidic peptide reduces significantly the synthesis of NO and NOS in brain of AD rat, but it dose not affect AChE activity remarkably. It is suggested that acidic peptide improves learning and memory of rat with Alzheimer disease probably by inhibiting the synthesis of toxic compound of NO or its toxicity.
