Aβ-,25-35-對海馬神經(jīng)元毒性作用的實驗研究.pdf

1、AB：淌對海馬神經(jīng)冠毒性作用的實驗研究詳細(xì)中文摘要，7背景：阿爾茨海默病(Alzheimer’sdisease，AD)是一種常見的老年認(rèn)知和記憶障礙疾病，臨床表現(xiàn)為進(jìn)行性智能減退和精神運動障礙。神經(jīng)元喪失、神經(jīng)纖維纏結(jié)(NFT)、老年斑(sP)和神經(jīng)元空泡樣變是AD的特征性病理變化，主要發(fā)生在前腦基底、海馬和大腦皮層，目前關(guān)于AD的發(fā)病機制尚不明了。研究資料提示，AD發(fā)病機制中的早期關(guān)鍵性環(huán)節(jié)是13淀粉樣前體蛋白(APP)的分解代謝、p

2、淀粉樣蛋白(Ap)的釋放與沉積。AB在神經(jīng)退變過程中的作用成為AD發(fā)病機制研究韻焦點。研究Ao對腦內(nèi)神經(jīng)元的毒性作用及其機制對闡明^AD的發(fā)病機制具有重要的意義。／目的：研究A13：。對大鼠學(xué)習(xí)和記憶活動的影響，探討At3：。：誘導(dǎo)海馬神經(jīng)細(xì)胞凋亡的作用機制，以及NGF和MK801的保護(hù)作用，進(jìn)一步闡明AB沉積。●。●。_。。1一與AD發(fā)病機制的相關(guān)性。／材料和方法：體內(nèi)實驗取20只SD大鼠隨機分成2組，A9處理組(實驗組)l和生理鹽水

3、組(對照組)，每組10只。采用立體定向法，給實驗組右側(cè)海馬CAl區(qū)微量注射Ap。，，每點3弘l(10pg／mD，對照組注入等體積生理鹽水。于注射前及注射后14天分別進(jìn)行避暗試驗作為行為檢鍘。于注射后15天每組隨機各取大鼠4只，經(jīng)粕多聚甲醛灌注后，取腦常規(guī)石蠟包埋。體外細(xì)胞培養(yǎng)取3天內(nèi)普通級新生SD大鼠30只，消毒取腦并取腦膜，無菌解剖液沖洗后分離海馬并剪碎，經(jīng)O25％胰雖白酶消化，將細(xì)胞接種于經(jīng)多聚賴氨酸(Lpolylysine)處理的

4、24孔細(xì)胞培養(yǎng)板中培養(yǎng)(培養(yǎng)液為神經(jīng)基礎(chǔ)培養(yǎng)液1327因子)，放置于37。C，5％二氧化碳培養(yǎng)箱中培養(yǎng)，將培養(yǎng)的細(xì)胞及切片組織進(jìn)行透射式電鏡檢測，觀察形態(tài)學(xué)改變。將培養(yǎng)的細(xì)胞及腦組織石蠟切片進(jìn)行P53、Bdxl、ACT、slooB免疫組化分析。將海馬細(xì)胞培養(yǎng)至第8天，加入不同濃度的ABlm和AB2Ⅻ(100pg／ml、lpg／ml、101x∥mli1001ag／m1)，孵育48小時，MTT檢測得到AB。。、A艮。，濃度依賴曲線。將AB。

5、。(10I‘∥d)和Ap：s。，(10弘∥fnl)分別孵育2h、12h、24h、36h、48h及對照，得到AB，。、郵：。時間依賴曲線。2ExperimentalStudyontheEffextofAB2摹3s蛐theToxicityofHippocampalNeuronAbstractinDetailBackground：Alzheimersdisease(AD)isthemostcommoncauseofdementiainolda

6、ge，itsmainmanifestationisprogressivecognitiveimpairmentLossofneuronsformationofneuroflbrillarytangle(NFT)andsenileplaque(sP)areitscharacteristicpathologicalchanges，whichmainlyoccllfinthenucleusbasalisofMeynert，hippocampu

7、sandcerebralcortexThepathogenesisofADstillremainsunclearStudydataindicatesthatthekeylinkinitsearlystageiscatabolismoff3Amyloidpreaarsorprotdn(APP)anddepositoff3Amyloidprotein(A8)TheroleofA13intheprocessofneurodegeneratio

8、nhasbecomethefocusofstudyonthepathogenesisofADStudy011theeffectofABonthetoxicityofneuronsofbrainanditsmechanismisofgreatimportancetothepathogenesisofADobiective：TostudytheeffectofAB25Ⅸonthelearningandmemoryactivitiesofra

9、tsandexplorethemechanismoftfippocampalneuronsdeathinducedbyA132535andtheprotectiveroleofnervegrowthfactorCNGF)andMKS01，inordertofurtherclarifytherelationshipbetweenA13depositandthepathogenesisofADMaterials＆methods：Invivo

10、experimem20SDratswerechooseanddividedintotwOgroupsrandomly：ABgroup(experimentalgroup)andnormalsalinegroup(controlgroulp)with10ratsineachgroupAB強3s39l(10咖1)wasstereotaxically兩ectedintotherighthippocampusCAlinexperimentalg

11、roupwithanegligibleforeachpointandtheequalanlotmtofnormalsalinewasinfusedintothes啪eareaincontrolgroupStepthroughtestwasconductedtochecklearningandmemoryactivhiesbefore船d14daysaftertheinjectionofAB2535ornormalsalinerespec

12、tively15daysaftertheinjection4ratswererandomlytakenfromeachgroupandthebrainsweredissectedandimbexldedwithparaffinforstudyInvitroexperiment30newbornSD瑚lswithin3daysatanordinarylevelweretakenandtheirbrainsweresterilizedina

13、septicanatomicalfluidThenhippocampuswasseparatedandCUtintopiecesWiththedigestionofO25％trypsasecellswereinoculatedontothecellcultureplaguewith24poresprocessedwithLpolyq3，sine(CUlturedfluidbeingNeuralBasialMediumB27factor)