[生物医药论文精品a]文冠果壳苷对侧脑室注射aβ1-42_致痴呆模型小鼠学习记忆障碍的改善作用

1文冠果壳苷对侧脑室注射A142致痴呆模型小鼠学习记忆障碍的改善作用邹莉波A1，王立华A2，迟天燕A1，纪雪飞A1，李伟A1，李晓春A1，付琪A1，杨柏珍A2（A0A3A4A5A6A7A8A9A10A11A7A9A12A10A13A14A6A9A15A16A4A5A0A0A17A17A0A18A19A20A3A21A7A15A4A5A10A22A23A24A25A16A4A5A0A0A17A17A0A18A26摘要目的本论文以侧脑室注射A142致痴呆小鼠为模型，考察了文冠果壳苷对痴呆模型小鼠学习记忆障碍的改善作用，并从抗氧化损伤角度探讨了文冠果壳苷对抗A142致痴呆作用的机制。方法采用小鼠新物体辨别、水迷宫、避暗等行为学测试方法研究文冠果壳苷改善学习记忆障碍的作用，用生化学方法检测脑组织总抗氧化能力（TAOC）、GSH/GSSG比值、MDA含量及CAT活性。结果文冠果壳苷008032MG/KG显著延长新物体辨别实验中对新物体探索时间并提高优先指数；显著缩G11713水迷宫试验中小鼠G2052G17810G4445G1852G2500的时间；显著G1955G4581避暗实验中的G19181G16835G8437数，延长G9520G1251G7411，缩G11713总G11017G1999时间。生化检测G13479果G15932G7138，文冠果壳苷显著提高模型小鼠G3835脑组织中CAT活性及TAOC，G1955G4581MDA含量、G3698G2164GSH含量并提高GSH/GSSG比值。结论G13520G990G6164G17860，文冠果壳苷能显著改善侧脑室注射A142致痴呆模型小鼠的学习记忆障碍，G1866作用机制G2499能G994对抗G14270G11013G3534损伤、G6245制G14038G17148G17819氧化G2465G5224等G7389G1863。关键词文冠果壳苷；A142；G19475G4584G14588G9035G21676G11161；抗氧化G19475G4584G14588G9035G21676G11161（ALZHEIMERSDISEASE,AD）的G2469G11161机制G3809G7446，G9053及G3822G12193G12082G13475G13007G13491G2163能障碍1，G13792G10628G7389的G8847G11115G14659物G13489G3835G3822数作用机制比G17751G2345G980，G3926ACHEG6245制G2070等，G1032G5214G11115G6940G7389G19492，G3252G8504，G4559G6226G1867G7389G13520G2524G8847G11115作用的抗痴呆新G14659G1185G7171G5415G1182G2319G14659G11040G7389G5465G6927G11784的G980G3835G19602G20076。文冠果XANTHOCERASSORBIFOLIABGE为G7092G5751G4388G12197文冠果G4658G7905物。G980G4658G980G12193，为G6117G3281G10317G7389G12193，G3324G6117G3281G1008G2283、华G2283、G16211G2283G18129G7389G7697G3533，G17176G9316G1028G4512。文冠果果G1173G994果壳G1147量比为108G794109，果壳G1147量G5468G3835，G3252G4590G7422G15999研究G2045用，G3534本G990G4658G5235G5335物。文冠果壳苷（XANTHOCERASIDE）G2375G7171G980G12193从文冠果果壳中提G2474的G989G14832G11350苷G12879化G2524物。G2081G7411研究G13479果G15932G71382，文冠果壳苷能G6338抗G1008莨菪碱、亚硝酸钠、40乙醇G6164致的记忆获得、巩固、再G10628和空间辨别障碍；显著改善侧脑室注射A2535致痴呆模型小鼠的学习记忆障碍，并显著提高模型小鼠脑内的SOD及GSHPX活性。本论文G3324G2081G7411工作的G3534础G990，进G980步考察文冠果壳苷对A142致痴呆模型小鼠学习记忆障碍的改善作用及机制，为文冠果壳苷开G2469成G8847G11115AD的新型G14659物提供依据。1实验材料11药品与试剂文冠果壳苷XANTHOCERASIDEG11013中G12197院生态G6164提供；银杏叶片深圳G9035王G14659业G7389G19492公司，批号20070704；AMYLOID142FRAGMENTG2283京博奥森生物技术G7389G19492公司；微量丙二醛MDA测试盒、氧化型谷胱甘肽GSSG及还原型谷胱甘肽GSH测试盒、G17819氧化氢酶CAT测试盒、总抗氧化能力T2AOC测试盒，均购G14270南京建成生物工程研究G6164。12实验动物昆G7138G12193小白鼠，雄性，体重2428G，G11013沈阳G14659G12197G3835学动物中心提供，G2524格证号SCXK（辽）2004008。13实验仪器722S分光光度计，G990G9035精密G12197学仪器G7389G19492公司；UV2000型荧光分光光度计，尤尼柯G990G9035仪器G7389G19492公司；JY92超声波细胞粉碎机及DY89G11017动玻璃匀浆机，宁波新芝生物G12197技G7389G19492公司；J2HS型低温超速离心机，美G3281BECKMAN；行为学实验装置小鼠避暗、水迷路、新物体辨别装置均为定制。2实验方法21动物模型的建立3及实验方案G2474雄性昆G7138G12193小白鼠，体重25G79430G。将小鼠随机分为6组，每组10只，包括假手术组，模型组，银杏叶片312MG/KG组，文冠果壳苷008、016、032MG/KG组。将小鼠用35的水G2524氯醛10ML/KG麻醉后，参考文献3A274A28A29A30除对照组外，每组小鼠均侧脑室G980G8437性注入5LG1969G13870态A142（含A142410PMOL，以G7092G14752生G10714G11428水G9354G16311，37G263120HG1363G1866成G1969G13870态），G11053G190361MIN，对照组小鼠G2528法侧脑室G13485G994等体G12227G9354G2070（1G715135的DMSO生G10714G11428水）。G17908模G2375G7097G17227开G3999G13485G14659，G14279行为学测试G13479G7475。G13485G14659后G1254489天进行新物体辨别实验，G125441014天进行水迷宫实验，G125441516天进行避暗实验，G7423G8437G13485G14659后，G7041G3848G2474脑匀浆，测定G2520G12193生化指G7643。22新物体辨别实验参照文献4进行。用测试G7411对新物体的优先指数G7481G2465G5224小鼠的记忆能力。新物体优先指数G7171探索新物体的时间G2356测试G7411探索总时间的G11346分比。23小鼠水迷宫法参照文献5进行。每只小鼠每天G16769G1346310G8437，间G1956030S。以30S内G2052G17810G4445G1852G2318为G8503G11842G2465G5224。记G5417小鼠G2052G17810G4445G1852G2318的时间，计G12651每天的G5191均时间和G8503G11842G2465G5224G8437数。24避暗法参照文献6进行。记G541724H后测验G7411G12544G980G8437进入暗室的G9520G1251G7411、5MIN内的G19181G16835G8437数和总G11017G1999时间，以G8504作为记忆成G13501。3实验结果31文冠果壳苷对侧脑室注射A142致痴呆模型小鼠学习记忆障碍的改善作用311水迷宫实验实验G13479果G3926G32821G6164G12046，G994G9354G2070对照组G11468比，模型组G3324水迷宫实验的G12544二天开G3999G2052G17810G4445G1852G2500的G9228G8903时间显著延长。G994模型组G11468比，文冠果壳苷G2520组G2070量依G17194性G3332缩G11713G2052G17810G4445G1852G2500的G9228G8903时间。3FIG1EFFECTOFXANTHOCERASIDEONSWIMMINGTIMEINWATERMAZETESTINMICEWITHICVA142N810,XSD312新物体辨别实验实验G13479果G15932G7138，G2520组小鼠G3324G16769G13463G7411对G1016G1022G11468G2528物体的探究时间G7092显著G5058G5334；测试G7411模型组小鼠对新物体和G7099物体的探究时间G1185G3534本G11468G2528，G13792G9354G2070对照组、银杏叶片组和文冠果壳苷032MG/KG组小鼠对新物体的探究时间显著延长；对新物体的优先指数显著G3698G2164，G994模型组G11468比G7389显著G5058G5334G16277G159321。TABLE1EFFECTOFXANTHOCERASIDEONPREFERENTIALINDEXFOROBJECT1ORNEWOBJECTINTHENOVELOBJECTRECOGNITIONINMICEWITHICVA142N1015,XSDP0001VSVEHICLEGROUPP0001VSMODELGROUPA31A32A33A31A33A32A34A31A34A32A35A31A35A32THESECONDDAYTHETHIRDDAYTHEFOURTHDAYTHEFIFTHDAYLATENCYSVEHICLEMODELYINXING008MG/KG016MG/KG032MG/KGTRAININGPHASETESTINGPHASEDOSEGROUPMG/KGPREFERENTIALINDEXFOROBJECT1PREFERENTIALINDEXFORTHENOVELOBJECTVEHICLE056006065006MODEL052016044006YINXING312049012064013XANTHOCERASIDE0080430050570070160460140610100320530120650134313避暗实验实验结果表明与溶剂对照组相比，模型组测试阶段潜伏期显著缩短，错误次数显著增加，总电击时间显著延长。与模型组相比，文冠果壳苷032、016MG/KG及银杏叶片组均能够显著延长潜伏期，并减少错误次数和总电击时间见表2。TABLE2EFFECTOFXANTHOCERASIDEONLATENCYINSTEPTHROUGHTESTINMICEWITHICVA142N1216,XSDP0001VSVEHICLEGROUPP005,P001VSMODELGROUP32文冠果壳苷抗氧化作用研究321文冠果壳苷对侧脑室注射A142致痴呆模型小鼠TAOC的影响实验结果如图2所示与溶剂对照组相比，模型组小鼠TAOC显著降低。与模型组比较，文冠果壳苷016MG/KG,032MG/KG组及银杏叶片组TAOC显著升高。A36A37A38A39A40A41A42A41A43A41A44A41A45A46A47A48A49A50A51A52A53A54A55A56A57A58A59A60A61A61A61A61A61A62A63A64A65A66A67A63A68A69A70A63A67A71A72A73A74A75A73A75A76A75A73A72A77A75A73A71A74A78FIG2EFFECTOFXANTHOCERASIDEONTHETAOCINTHEBRAINOFMICEWITHICVA142N810,XSDP005VSVEHICLEGROUPP005,P001VSMODELGROUPXANTHOCERASIDEYINXINGMG/KGGROUPDOSEMG/KGLATENCYSVEHICLE2240711725MODEL49539540YINXING3121356412095XANTHOCERASIDE0081254312406016184801244603218747115525322文冠果壳苷对侧脑室注射A142致痴呆模型小鼠GSH、GSSG含量的影响实验结果如图3、图4所示，与溶剂对照组相比，模型组小鼠脑组织GSSG含量未见显著改变，但GSH含量及GSH/GSSG显著降低。与模型组比较，文冠果壳苷016MG/KG，032MG/KG组及银杏叶组小鼠脑组织GSSG含量未见显著改变，但GSH含量及GSH/GSSG显著升高。A79A80A79A81A79A79A81A80A79A82A79A79A82A80A79A83A84A85A86A87A88A84A89A90A91A84A88A92A81A93A82A79A93A79A94A79A93A81A95A79A93A92A82A96A97A98A99A100A101A102A103A101A102A104A105A106A100A107A108A109A110A111A112A112A112FIG3EFFECTOFXANTHOCERASIDEONTHECONTENTOFGSHINTHEBRAINOFMICEWITHICVA142N810,XSDP005VSVEHICLEGROUPP005VSMODELGROUP005115225335VEHICLEMODEL312008016032GSH/GSSGFIG4EFFECTOFXANTHOCERASIDEONTHERATIOOFGSH/GSSGINTHEBRAINOFMICEWITHICVA142N810,XSDP001VSVEHICLEGROUPP005,P001VSMODELGROUP323文冠果壳苷对侧脑室注射A142致痴呆模型小鼠过氧化氢酶CAT活力的影响XANTHOCERASIDEYINXINGMG/KGXANTHOCERASIDEYINXINGMG/KG6实验结果如图5所示与溶剂对照组相比，模型组小鼠CAT活力显著降低。与模型组比较，文冠果壳苷032MG/KG组，银杏叶片组CAT活力显著升高。A113A114A113A113A115A113A113A116A113A113A117A118A118A119A118A118A118A119A120A118A118A119A121A118A118A119A122A118A118A119A117A118A118A120A118A118A118A123A124A125A126A127A128A124A129A130A131A124A128A132A119A133A120A118A133A118A117A118A133A119A122A118A133A132A120A134A135A136A135A137A138A139A140A139A138A141A142A143A144A145A146A147A148A138A149A150A150A151A151FIG5EFFECTOFXANTHOCERASIDEONTHECATACTIVITYINTHEBRAINOFMICEWITHICVA142N810,XSDP001VSVEHICLEGROUPP005VSMODELGROUP324文冠果壳苷对侧脑室注射A142致痴呆模型小鼠MDA含量的影响实验结果如图6所示与溶剂对照组相比，模型组小鼠MDA含量显著升高。与模型组比较，文冠果壳苷008MG/KG,016MG/KG,032MG/KG组及银杏叶片组MDA含量显著降低。A152A153A154A155A156A157A152A157A153A157A154A157A155A157A156A158A159A160A161A162A163A164A165A163A164A166A163A167A162A168A169A167A170A171A172A162A164A173A174A175A175A175A175A176A177A178A179A180A181A177A182A183A184A177A181A185A186A187A188A189A187A189A190A189A187A186A191A189A187A185A188FIG6EFFECTOFXANTHOCERASIDEONTHECONTENTOFMDAINTHEBRAINOFMICEWITHICVA142N810,XSDP005VSVEHICLEGROUPP005,P001VSMODELGROUPXANTHOCERASIDEYINXINGMG/KGXANTHOCERASIDEYINXINGMG/KG74讨论有研究表明，氧自由基与A神经毒性有关，应用多光子成像技术，在AD转基因鼠及AD患者的体内外实验都提示A沉积与氧自由基的产生有直接关系，A沉积所形成的淀粉样斑块G13870G19610G10301内有G3835量的氧自由基产生。A的神经毒性G1328用所G14280的G13466G14002氧自由基G6451G1272G1039G16213有G1209G991G17896G5464G311AG7424G17535G2499G1328G1038氧自由基G1391体，产生活性氧G727G312AG2499G17902G17819G16837G4560G1363氧自由基产生G727G313AG17902G17819与G10317G5334的G2475体结G2524G9620活小G14026G17148G13466G14002，产生G3835量自由基，加G2107氧G2282应G9620。氧自由基对A的G13870G19610G1075有G18337G16213G1328用，AG13969段的G7588G1135G8700基G18252G7143G2475自由基G6927击，并G1363G15519G11345G17148G2010子G1144G13864G727A的G1254435G1313G8700基G18252G15519G8700G18252MET35在G11107G8712G2318的G1025G18108，氧G2282的MET35含有G7368G7509性G2282的基G3254，G1363AG13870G19610加G17907。AG1146G17902G17819G5445G2721脑G12373G16314G2530G14192G990G14158苷G18252G10627G2282G18250的活性,G16855G14422GG15519G11345G1610G13864的G1461G2507G1268G4560G5445G2721氧自由基G1207G16886。G13520G990所G17860，AD脑组织G1025氧自由基G1207G16886G2469生G13022G1093，AG2499G1209G11487G1328G7171氧G2282应G9620与AD脑神经G1815G8527G1141G1055间的G1610G13864G2010子7。G6117G1216G2081期研究结果表明，文冠果壳苷能够G6338G6251G1008G14716G14782G11909、G1134G11825G18252G19060、40G1069G18267所G14280的G16772G5530G14731G5483、G5053G3278、G1889G10628和G12366间G17788G2047G19568G11873G727显著改G2904G1403脑G4472G8892G4568A2535G14280G11208呆模型小鼠的学习G16772G5530G19568G11873，并显著提高模型小鼠脑内的SOD及GSHPX活性。G7424实验进一步证明文冠果壳苷在微克级的剂量G991能够显著改G2904A142G14280G11208呆模型小鼠G12366间学习G16772G5530G19568G11873及G10301体G17788G2047G16772G5530G19568G11873G727显著降低脑组织G1025MDA含量，提高GSH/GSSG比值，增加总G6251氧G2282能力，提高CAT活力。G990G17860结果提示，文冠果壳苷改G2904A142G14280G11208呆模型小鼠学习G16772G5530G19568G11873的G1328用G2499能与其G6251自由基G6451G1272有关。参考文献1PARIHARMS,HEMNANITALZHEIMERSDISEASEPATHOGENESISANDTHERAPEUTICINTERVENTIONSJJOURNALOFCLINICALNEUROSCIENCE,2004,1154564672LIUX,YANGXA,QUC,ETALEFFECTSOFEXTRACTOFTHEHUSKOFXANTHOCERASSORBIFOLIABUNGEONLEARNINGANDMEMORYDEFICITSJTRADITIONALCHINESEDRUGRESEARCHANDCLINICALPHARMACOLOGYA192A193A194A193A195A196A197A198A199A200A201A202A202A203A204A205A206A207A205A208A209A201A210A211A201A2123JINHJ,KIMBHC,TNABESHIMA,ETALAMYLOID142INDUCEDLEARNINGANDMEMORYDEFICITSINMICEINVOLVEMENTOFOXIDATIVEBURDENSINTHEHIPPOCAMPUSANDCEREBRALCORTEXJBEHAVIOURALBRAINRESEARCH20041551851964FABIANFERNANDEZ,CRAIGCGARNEROBJECTRECOGNITIONMEMORYISCONSERVEDINTS1CJE,AMOUSEMODELOFDOWNSYNDROMEJNEUROSCIENCELETTERS,2007,4211371415XIANYFA204QUC,WUZ,ETALCOMPARISIONOFTHEEFFECTSOFCHPQSANDCAPQSONLEARNINGANDMEMORYJJOURNALOFSHENYANGPHARMACEUTICALUNIVERSITYA213A214A215A216A217A218A219A219A220A2212006A222A2231A22443476XIANY,JIXA225LIUX,ETALEXPERIMENTALRESEARCHONIMPROVINGLEARNINGANDMEMORYABILITIESOFTOTALSTERONEEXTRACTSOFRHAPONTICUMUNIFLORUMA226A227A228TRADITIONALCHINESEDRUGRESEARCHANDCLINICALPHARMACOLOGYA229A216A230A216A231A232A233A216A234A2212005,1664054087CROUCHPJ,HARDINGSMEWHITEAR,ETALMECHANISMOFAMEDIATEDNEURODEGENERATIONINALZHEIMERSDISEASEJTHEINTERNATIONALJOURNALOFBIOCHEMISTRY2INSTITUTEOFAPPLIEDECOLOGY,CHINESEACADEMYOFSCIE