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1、Jul.200521(4)365～371神经解剖学杂志(ChineseJournalofNeuroanatomy)BDNF和神经干细胞联用对老年痴呆鼠基底前脑①胆碱能神经元和学习记忆能力的影响3宣爱国龙大宏杨丹迪谷海刚冷水龙(广州医学院解剖学教研室,广州510182)摘要探讨脑源性神经营养因子(BDNF)和神经干细胞(NSCs)联合应用对老年痴呆大鼠基底前脑胆碱能神经元及大鼠学习记忆能力的影响。利用无血清培养技术获得新生SD鼠的海马NSCs,行BrdU标记。切断SD大鼠左侧穹隆海马伞,基底前脑注射NSCs,同时侧脑室注射BDNF,2周后行nestin和BrdU/NF、BrdU/GFAP免
2、疫荧光双标染色,4周后行Y迷宫测试,免疫组化结合图像分析技术观察各组大鼠基底前脑神经营养因子受体(NGFR)阳性神经元数目变化。结果发现,移植后NSCs能够在宿主体内存活且分化成神经元和胶质细胞;免疫组化结果显示损伤组大鼠胆碱能神经元数在内侧隔阂(MS)和斜角带(VDB)分别减少64.3%和49.3%,较正常组明显下降(P<0.01);移植组神经元数下降34.1%和27.6%较损伤组有改善(P<0.05),但与正常组比较有显著性差异(P<0.05);联合组神经元数减少15.1%和18.6%,与正常组无显著性差异(P>0.05)。Y迷宫测试结果显示,大鼠的学习记忆能力与基底前脑NGFR阳性
3、细胞数呈正相关。提示,BDNF和NSCs移植的联用较单独使用NSCs或BD2NF更好地改善老年痴呆大鼠的学习记忆能力。关键词老年性痴呆脑源性神经营养因子神经干细胞胆碱能神经元学习记忆大鼠THEEFFECTOFBDNFANDNEURALSTEMCELLSUNITEAPPLICATIONONCHOLINERGICNEURONSOFTHEBASALFOREBRAININMODELRATSWITHSENILEDEMENTIAANDABILITIESOFLEARNINGANDMEMORYXuanAiguo,LongDahong,YangDandi,GuHaigang,LengShuilong(De
4、partmentofAnatomy,GuangzhouMedicalCollege,Guangzhou510182)AbstractTostudytheeffectofbrainderivedneurotrophicfactor(BDNF)andneuralstemcells(NSCs)uniteapplicationonthecholiner2gicneuronsinbasalforebrainandleaningandmemoryinratswithAlzheimer`sdisease.NSCswereobtainedfromhippocampusbyusingtheserum2fr
5、eeculturing.NSCslabeledbyBrdUweretransplantedintothetransectedbasalforebrain,thenBDNFwasinjectedintolateralven2tricle.After2weeks,Nestinstaining,BrdU/NFandBrdU/GFAPdoublelabeledimmunofluorescencewereusedtodetectthesurvivalandtheneuronaldifferentiationoftheBrdUlabeledcellsinthebasalforebrain.After
6、4weeks,learningandmemoryabilitiesweremeasuredbyY2mazetest.Thenumberofbasalforebraincholinergicneuronswasanalyzedbyusingimmunohistochemicalmethodcombinedwithtech2niqueofmicromeasureandimageanalysis.ThedoublelabeledNSCswereobservedinthebasalforebrainanddifferentiatedintoneuronsandastrocytes.Thenumb
7、erofcholinergicneuronsforinjuredgroupswasdecreasedby64.3%and49.3%respectivelyinmedialseptum(MS)andverticaldiagonalbranch(VDB),significantdifferencewasfoundbetweentheinjuredgroupsandnormalcontrolgroups(P<0.01).Thenumber
