碱性成纤维细胞生长因子对血管性痴呆大鼠海马胆碱能神经元的影响

目的皮下注射碱性成纤维细胞生长因子(bFGF)于血管性痴呆大鼠,研究用药前后大鼠海马胆碱能神经元的变化。方法制作血管性痴呆(VD)大鼠模型,随机取用VD大鼠模型12只,分治疗组6只,痴呆组6只。另外,取假手术组6只。皮下注射bFGF于治疗组中血管性痴呆大鼠。治疗5周后,以Morris水迷宫定位航行试验和空间探索试验来检测大鼠的学习记忆能力,乙酰胆碱转移酶(ChAT)免疫组织化学染色观察海马CA1区胆碱能神经元数目的变化。结果治疗组大鼠海马CA1区胆碱能神经元数目较痴呆组明显增多。结论皮下注射bFGF后能迁移至海马,诱导海马产生具有ChAT活性神经元,所产生的ChAT活性神经元可能就是胆碱能神经元。     

银丹心脑通软胶囊对血管性痴呆大鼠行为学及海马组织乙酰胆碱酯酶的影响

目的 观察银丹心脑通软胶囊对血管性痴呆大鼠学习记忆及海马组织乙酰胆碱酯酶活性的影响,探讨银丹心脑通软胶囊治疗血管性痴呆的作用机制. 方法 采用反复夹闭双侧颈总动脉再灌注、同时腹膜腔内注射硝普钠方法制作拟血管性痴呆大鼠模型.将造模成功的20只大鼠按随机数字表法分为血管性痴呆组及银丹心脑通软胶囊治疗组(各10只),另以条件匹配的10只大鼠为假手术组.于造模结束大鼠苏醒后30min以及处死前测定大鼠神经功能评分,采用跳台实验检测大鼠学习记忆能力,采用放射免疫法检测海马组织乙酰胆碱酯酶活性. 结果 大鼠处死前,与假手术组比较,血管性痴呆组和银丹心脑通软胶囊治疗组大鼠神经功能评分仍明显增高,差异有统计学意义(P＜0.05);银丹心脑通软胶囊治疗组大鼠神经功能评分明显低于血管性痴呆模型组,差异有统计学意义(P＜0.05).与血管性痴呆组比较,银丹心脑通软胶囊治疗组大鼠反应时间明显缩短,潜伏时间明显延长,错误次数明显减少,差异均有统计学意义(P＜0.05).银丹心脑通软胶囊治疗组海马组织AChE活性明显低于血管性痴呆组,差异有统计学意义(P＜0.05). 结论 银丹心脑通软胶囊能增强血管性痴呆模型动物的学习记忆功能,其作用机制可能与抑制乙酰胆碱酯酶活性有关.     

血管性痴呆患者血中胆碱酯酶活性及神经型尼古丁受体mRNA表达水平变化

目的 探讨血管性痴呆(vascular dementia,VD)患者血中胆碱酯酶活性及神经型尼古丁乙酰胆碱能受体(nicotinic acetyleholine receptor,nAchR)mRNA表达的变化.方法 分组比较测定59例血管性痴呆、33例正常老年人空腹血浆胆碱酯酶浓度及血液白细胞nAchRa4、β2亚单位mRNA表达水平.结果 血管性痴呆组与对照组乙酰胆碱酯酶活性分别为(1.70±1.17nmol/min/ml,2.18±0.87nmol/min/ml),VD组与对照组相比差异有显著性(P<0.05);VD组与对照组丁酰胆碱酯酶活性分别为(61.08±38.23nmol/min/ml,70.41 28.82nmol/min/ml),相比差异无显著性(P<0.05).乙酰胆碱酯酶及丁酰胆碱酯酶均无性别的差异;VD患者血液白细胞nAchR a4亚单位mRNA表达水平(0.926±0.411)较正常对照组(1.41±0.22)降低(P<0.05);VD患者nAchR β2亚单位mR-NA表达水平(2.14±0.18)与正常对照组(2.19±0.16)差异无显著性(P<0.05).结论 血管性痴呆患者血浆乙酰胆碱酯酶活性和血液白细胞中nAehR α4亚单位mRNA表达水平较正常对照老年人降低,但丁酰胆碱酯酶和nAeh β2 mRNA无改变,这些变化可能对血管性痴呆的诊断有一定临床意义.     

天智颗粒对血管性痴呆大鼠行为学及脑内AchE、ChAT活性的影响

目的观察天智颗粒对血管性痴呆大鼠行为学习记忆能力和乙酰胆碱代谢酶活性的影响,从而探讨天智颗粒改善记忆的机制.方法采用双侧颈总动脉结扎法建立血管性痴呆模型,治疗组用天智颗粒(5 g/kg)灌胃,模型组和假手术组用同体积的蒸馏水代替灌胃,1次/天,对照组未做任何处理.30 d后,采用三等分Y型电迷宫和比色法分别测定皮质、海马区乙酰胆碱脂酶(AchE)活性和基底前脑区胆碱乙酰转移酶(ChaT)活性.观察、比较各组大鼠行为学和乙酰胆碱代谢酶的变化规律和差异.结果治疗30d后,治疗组与模型组相比,学习记忆能力和基底前脑ChaT活性明显提高(P＜0.05),但皮质和海马区AchE活性变化无统计学意义.结论天智颗粒能明显促进血管性痴呆大鼠学习记忆能力和基底前脑区ChaT活性,其药理机制可能和改变乙酰胆碱代谢酶活性有关.     

脐血干细胞海马移植对VD大鼠脑内Ach及AchE活性的影响

目的观察海马移植脐血干细胞对血管性痴呆(VD)大鼠脑内Ach含量及AchE活性的影响。方法分离正常分娩胎儿的脐带血单个核细胞,以贴壁培养法得到间质干细胞(MSCs),采用RT-PCR方法检测Nestin及musashi-1。制作VD大鼠模型,8～10d后行脐血干细胞海马移植。移植4周后,检测大鼠的学习记忆能力,并检测其脑内乙酰胆碱(Ach)含量及乙酰胆碱脂酶(AchE)活性。结果脐血干细胞在分离后48h高表达Nestin及musashi-1;海马移植脐血干细胞4周后,治疗组与模型组比较,Ach含量及AchE活性明显升高(P<0.01),大鼠学习记忆能力增强。结论脐血中表达神经干细胞(NSCs)的标志物,海马移植脐血干细胞能明显改善VD大鼠胆碱能系统的功能,调节脑内Ach生理代谢,增强学习记忆能力,从而达到治疗VD的目的。     

银丹心脑通软胶囊对血管性痴呆模型大鼠行为学及脑组织自由基代谢的影响

目的 研究银丹心脑通(YDXNT)软胶囊对血管性痴呆(VD)模型大鼠学习记忆能力及超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量的影响. 方法 30只SD大鼠按照随机数字表法分成3组:假损伤组、VD模型组、YDXNT治疗组.后2组采用血管闭塞法复制VD模型,假损伤组除不结扎双侧颈总动脉外,其余处理与VD模型组、YDXNT治疗组相同.大鼠术后当天清醒后开始灌胃给药,YDXNT治疗组参照人的给药剂量,每只每次灌服lrnL液体(预先取YDXNT胶囊9粒去除胶囊溶解于360 mL生理盐水中配成液体备用),3次/d,连续注射4周.假损伤组与VD模型组均灌胃等体积生理盐水.Morris水迷宫检测大鼠学习记忆能力,放射免疫方法检测海马组织SOD活性与MDA含量. 结果 YDXNT治疗组大鼠找到平台的时间较VD模型组明显缩短,找到平台的游泳距离较VD模型组明显减少,差异均有统计学意义(P＜0.05).与VD模型组大鼠比较,YDXNT治疗组大鼠SOD活性明显升高,MDA含量明显降低,差异均有统计学意义(P＜0.05). 结论 YDXNT胶囊可增加脑组织氧自由基代谢能力,改善VD大鼠学习记忆能力.     

盐酸美金刚对血管性痴呆大鼠疗效观察

目的 观察盐酸美金刚对血管性痴呆大鼠的疗效及作用机制.方法 80只健康Wistar大鼠(月龄12～14个月),体质量300～400 g,随机分为对照组、模型组、美金刚对照组及美金刚治疗组,每组20只.采用持久双侧颈总动脉结扎术造成血管性痴呆大鼠模型,美金刚对照组及美金刚治疗组于术后8周开始以美金刚(5 mg·kg-1)每天灌胃,对照组和模型组以同等量的0.5 g/L羧甲基纤维素钠灌胃,连续4周.采用Morris水迷宫衡量大鼠学习记忆水平;测定大鼠脑皮层、海马组织乙酰胆碱酯酶(AChE)、胆碱乙酰转移酶(ChAT)、丙二醛(MDA)、谷胱甘肽(GSH)活性的变化.结果 术后12周,与对照组相比,模型组大鼠的学习记忆能力明显下降(P<0.05);脑皮层、海马组织内AChE活性明显升高(P<0.05),ChAT活性明显降低(P<0.05),MDA活性明显升高(P<0.05),GSH活性明显降低(P<0.05);与模型组相比,美金刚治疗组大鼠的学习记忆能力明显提高(P<0.05);脑皮层、海马组织内AChE活性及ChAT活性差异无统计学意义(P>0.05)、MDA活性明显降低(P<0.05)、GSH活性明显升高(P<0.05).结论 盐酸美金刚对血管性痴呆大鼠的学习记忆能力有明显提高,作用机制是通过调节脑组织内MDA及GSH的活性来实现的,该实验研究为临床上血管性痴呆的治疗提供实验基础及理论依据.     

盐酸多奈哌齐对血管性痴呆模型大鼠大脑NO及iNOS的影响

目的：观察盐酸多奈哌齐（DP）对血管性痴呆模型大鼠iNOS表达的影响,探讨其抗血管性痴呆的作用及机制。方法将SD大鼠随机分为假手术组、实验对照组、DP治疗组,每组大鼠20只,采用双侧颈总动脉结扎法制作血管性痴呆大鼠模型。DP干预治疗后,用“Y”型迷宫检测大鼠学习记忆能力,比色法检测大鼠大脑NO及NOS的表达。结果与实验对照组比较,DP治疗后VD大鼠学习记忆能力明显提高,海马iNOS表达明显减少（P＜0.01）。结论盐酸多奈哌齐可改善学习记忆能力,其机制可能是通过下调iNOS的过度表达,减少NO的生成,抑制神经细胞损伤。     

bFGF促进痴呆大鼠学习记忆能力恢复及可能机制

目的　探讨碱性成纤维细胞生长因子 (b FGF)对痴呆大鼠学习记忆能力的影响 ,探讨其可能机制。方法　痴呆组、痴呆治疗组及痴呆对照组大鼠前脑 Meynert基底核注射红藻氨酸制造痴呆模型 ,正常对照组同法注射生理盐水 ;痴呆治疗组造模后 30 min、1d、3d、5 d及 7d侧脑室注射 b FGF,痴呆对照组同时间注射生理盐水 ,造模30 d后正常对照组、痴呆组、痴呆对照组及痴呆治疗组大鼠通过 Y迷宫进行学习记忆能力测试、乙酰胆碱酯酶(Ach E)细胞化学染色及尼氏染色 ,并测量基底前脑皮层及海马各区 Ach E纤维密度。结果　痴呆组大鼠学习及记忆能力较正常对照组减低 (P<0 .0 1) ,基底前脑皮层及海马各区 Ach E纤维密度减低 (P<0 .0 1)。治疗组 Y迷宫学习及记忆能力较痴呆对照组有改善 (P<0 .0 1) ,基底前脑皮层及海马各区 Ach E纤维密度亦有所增加 (P<0 .0 1) ,但是没达到正常对照组的水平 (P<0 .0 1)。结论　 b FGF可改善痴呆模型大鼠 Y迷宫学习记忆能力 ;脑内 Ach E纤维密度提高及海马神经元密度增加是 b FGF作用产生的可能机制。     

康复训练对血管性痴呆大鼠认知功能障碍的影响及其分子机制

目的探讨康复训练对血管性痴呆大鼠空间学习记忆能力的改善作用,并从分子学角度探讨其机制。方法选择Wistar雄性大鼠25只,随机分为正常对照组(5只)、血管性痴呆组(10只)和康复训练组(10只)。采用双侧颈总动脉阻断法制作血管性痴呆大鼠模型,康复训练组大鼠于手术后2d开始进行康复训练;用Westernblotting方法测定大鼠海马组织中N-甲基-D-天冬氨酸受体1含量,用长时程增强及Morris水迷宫试验评价大鼠空间学习记忆能力。结果Morris水迷宫试验显示,血管性痴呆组大鼠水迷宫隐匿平台逃避潜伏期明显延长,经康复训练后学习记忆能力有所改善,但与正常对照组相比差异仍有统计学意义(P<0.05)。发生血管性痴呆后,大鼠海马组织中N-甲基-D-天冬氨酸受体1含量明显降低,康复训练使其含量较训练前明显升高,但仍未能达到正常对照组水平(P<0.05);血管性痴呆大鼠的长时程增强诱导功能亦明显受到抑制,康复训练后虽明显改善但未恢复至正常水平(P<0.05)。结论康复训练可提高大鼠空间学习记忆能力,促进长时程增强的形成,其分子机制可能与海马组织中N-甲基-D-天冬氨酸受体1表达水平的增高有关。     

海马锥体细胞的变化对血管性痴呆大鼠学习记忆能力的影响

目的研究海马锥体细胞的变化对血管性痴呆大鼠学习记忆能力的影响。方法将大鼠随机分成手术组及对照组,手术组制作血管性痴呆大鼠模型,30d后用Morris水迷宫检测两组大鼠学习记忆能力及HE染色检测海马锥体细胞数目的变化。结果手术组大鼠学习记忆能力明显下降,海马锥体细胞数目较对照组明显减少。结论血管性痴呆大鼠海马锥体细胞减少,学习记忆能力下降。     

Basic fibroblast growth factor improves learning and memory deficits in a mouse model of vascular dementia Associated with the role of free radicals clearance?

BACKGROUND:Basic fibroblast growth factor(bFGF)exhibits neuroprotective functions,but the possible mechanisms of bFGF on vascular dementia remain unclear.OBJECTIVE:To explore the neureprotective effects of bFGF on a mouse model of vascular dementia,with focus on oxidative damage.DESIGN,TIME AND SETTING:A randomized,controlled,animal experiment was performed at the Medical College of Beihua University from March to December 2008.MATERIALS:bFGF was purchased from Peprotech,USA.METHODS:A total of 80 healthy,Kunming mice were randomly assigned to control,sham-surgery,model,and bFGF groups.The model and bFGF groups were used to establish vascular dementia models by repetitive cerebral ischemia-reperfusion in a conscious state.In addition,bFGF group mice were intraperitoneally injected with bFGF(100 μg/kg)following model establishment,once a day for 7 consecutive days.MAIN OUTCOME MEASURES:The Morris water maze was used to determine the influence of bFGF on learning and memory abilities in vascular dementia mice.The pathomorphological changes in hippocampal CA1 neurons were observed by Nissl staining.Superoxide dismutase and malondialdehyde changes were analyzed using biochemical analysis methods.Annexin V-FITC/PI-double-labeled flow cytometry was used to detect neuronal apoptosis.RESULTS:Learning and memory functions in model mice significantly decreased,as characterized by prolonged latency and reduced time and number of platform crossings(P < 0.01,P < 0.05).Superoxide dismutase activity was significantly reduced,malondialdehyde content was significantly increased(P < 0.01),and hippocampal neuronal apoptosis was increased(P < 0.01)following vascular dementia,bFGF increased superoxide dismutase activity,decreased malondialdehyde content,and reduced hippocampal neuronal apoptosis(P < 0.01),which resulted in improved learning and memory in mice with vascular dementia.CONCLUSION:bFGF improved learning and memory deficits in mice with vascular dementia by reducing free radical injury and inhibiting hippocampal neuronal apoptosis.     

A ginkgo biloba extract promotes proliferation of endogenous neural stem cells in vascular dementia rats

The ginkgo biloba extract EGb761 improves memory loss and cognitive impairments in patients with senile dementia. It also promotes proliferation of neural stem cells in the subventricular zone in Parkinson’s disease model mice and in the hippocampal zone of young epileptic rats. However, it remains unclear whether EGb761 enhances proliferation of endogenous neural stem cells in the brain of rats with vascular dementia. In this study, a vascular dementia model was established by repeatedly clipping and reperfusing the bilateral common carotid arteries of rats in combination with an intraperitoneal injection of a sodium nitroprusside solution. Seven days after establishing the model, rats were intragastrically given EGb761 at 50 mg/kg per day. Learning and memory abilities were assessed using the Morris water maze and proliferation of endogenous neural stem cells in the subventricular zone and dentate gyrus were labeled by 5-bromo-2-deoxyuridine immunofluorescence in all rats at 15 days, and 1, 2, and 4 months after model establishment. The escape latencies in Morris water maze tests of rats with vascular dementia after EGb761 treatment were significantly shorter than the model group. Immunofluorescence staining showed that the number and proliferation of 5-bromo-2-deoxyuridine-positive cells in the subventricular zone and dentate gyrus of the EGb761-treated group were significantly higher than in the model group. These experimental findings suggest that EGb761 enhances proliferation of neural stem cells in the subventricular zone and dentate gyrus, and significantly improves learning and memory in rats with vascular dementia.     

Effect of electroacupuncture on the expression of mTOR and eIF4E in hippocampus of rats with vascular dementia

Clinically, electroacupuncture is proved to be an effective therapy for vascular dementia; however, their mechanisms remain uncertain. The aim of the current study was to investigate the mechanism of electroacupuncture therapy for vascular dementia. One month after a vascular dementia animal model was established by bilateral occlusion of common carotid arteries, electroacupuncture treatment was given at “Baihui” (DU20), “Dazhui” (DU14), and “Shenshu” (BL23). Morris water maze was used to assess the learning and memory ability of rats. Western blot assay was performed to detect the expression of mammalian target of rapamycin (mTOR) and eukaryotic translation initiation factor 4E (eIF4E) in hippocampus of rats. Morris water maze test showed that electroacupuncture improved the learning ability of vascular dementia rats. Western blot assay revealed that the expression level of mTOR and eIF4E in the electroacupuncture group and sham-operated group was higher than that in the vascular dementia group (P < 0.05). In conclusion, the decreasing expression of mTOR and eIF4E plays important roles in the pathogenesis of vascular dementia. Electroacupuncture improves learning and memory ability by up-regulating expression of mTOR and eIF4E in the hippocampus of vascular dementia rats.     

黄精口服液对血管性痴呆大鼠学习记忆的影响

目的：观察黄精口服液对血管性痴呆大鼠的干预效果。方法：采用永久性结扎双侧颈总动脉法建立血管性痴呆动物模型。应用Morris水迷宫检测大鼠学习记忆能力。结果：术后3．5个月(给药2．5个月)痴呆 黄精口服液组大鼠平均逃避潜伏期较血管性痴呆组和痴呆 生理盐水组大鼠明显缩短。结论：黄精口服液可改善血管性痴呆大鼠学习记忆能力。     

Effects of electroacupuncture on the expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in the hippocampus of rats with vascular dementia

This study investigated the mechanism underlying electroacupuncture therapy for vascular dementia through electroacupuncture at the acupoints of Baihui (DU20), Dazhui (DU14), and bilateral Shenshu (BL23) in a rat model of vascular dementia produced by bilateral middle cerebral artery occlusion. Morris water maze test showed that electroacupuncture improved the learning ability of vascular dementia rats. Western blot assay revealed that the expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in vascular dementia rats was significantly increased after electroacupuncture, compared with the model group that was not treated with acupuncture. The average escape latency was also shortened after electroacupuncture, and escape strategies in the spatial probe test improved from edge and random searches, to linear and trending swim pathways. The experimental findings indicate that electroacupuncture improves learning and memory ability by up-regulating expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in the hippocampus of vascular dementia rats.     

甲基维生素B12对拟血管性痴呆大鼠记忆功能及脑白质病理变化的影响

目的 观察甲基维生素B12对拟血管性痴呆大鼠记忆功能及脑白质病理变化的影响。方法 反复前脑缺血制备大鼠拟血管性痴呆模型，造模后甲基维生素B12治疗1周、2周、4周时Morris水迷宫检测大鼠学习记忆功能、LFB染色观察白质髓鞘变化、电镜下观察少突胶质细胞、髓鞘超微结构，并与假手术组及对照组对比。结果 治疗4周大鼠游迷宫时间明显短于对照组，造模2周、4周时LFB染色均可见白质髓鞘脱失，但治疗组明显较对照组轻。造模4周电镜下少突胶质细胞、髓鞘改变治疗组轻于对照组。结论 甲基维生素B12能改善拟血管性痴呆大鼠记忆功能，并能减轻脑白质病理变化。