远志总皂苷对癫痫模型大鼠学习记忆及海马LTP的影响

目的观察远志总皂苷(TEN)对戊四氮(PTZ)点燃癫痫模型大鼠空间学习记忆能力及海马长时程增强(LTP)的影响。方法雄性Wister大鼠随机分为对照组、癫痫模型组、TEN组,利用水迷宫法观察各组大鼠的空间学习记忆能力,利用电生理学方法通过对高频刺激前后海马CA1区场兴奋性突触后电位(fEPSP)幅度比较检测大鼠海马LTP变化。结果与对照组相比较,癫痫模型组空间学习记忆能力显著下降,海马CA1区LTP受到显著抑制(P0.05);TEN可改善上述指标的异常变化(P0.05)。结论TEN可改善癫痫模型大鼠空间学习记忆能力及海马LTP的抑制。     

小牛血清去蛋白注射液对致痫实验大鼠的影响

目的探讨小牛血清去蛋白注射液（DCBSI）对戊四唑（PTZ）诱导的大鼠癫痫的作用及其机制。方法健康SD大鼠38只,随机分为5组,对照组、癫痫组和DCBSI 3个剂量组（20、40、80 mg/kg）。观察大鼠腹腔注射PTZ前后癫痫发作的情况,按Racine分级标准记录,同时记录皮层脑电图（ECOG）,观察癫痫样放电的潜伏期及1 h内癫痫样放电活动持续时间。应用Western blot电泳分析海马内CA1区P38促分裂原活化蛋白激酶（MAPK）及其上游MKK3、下游c-MYC相关蛋白的表达。结果对照组给予PTZ后均出现癫痫发作,程度均为5级;DCBSI组发作程度明显减轻。ECOG潜伏期延长,痫样放电1 h持续时间缩短。与对照组相比,癫痫组和DCBSI 3个剂量组大鼠CA1区蛋白表达强度升高（P〈0.05）;与癫痫组相比,DCBSI 3个剂量组大鼠CA1区蛋白表达强度降低（P〈0.05）。结论 DCBSI对PTZ诱导的癫痫大鼠的发作有明显的对抗作用,对癫痫大鼠海马具有保护作用,其作用与其抑制P38MAPK通路相关蛋白的表达相关。     

戊四氮致痫大鼠脑组织NT-4的变化及百里香与薄荷精油的干预作用

目的观察戊四氮致痫大鼠脑组织神经营养因子-4（Neurotrophin-4,NT-4）的变化,利用百里香与薄荷精油制备雾化吸入剂进行干预治疗,研究癫痫的发病机理及精油的作用机制。方法健康雄性Wistar实验大鼠40只,随机分为正常对照组、癫痫模型组、精油治疗I组、精油治疗Ⅱ组、每组10只。模型组和治疗组均用致痫亚惊厥剂量的戊四氮（PTZ）腹腔注射制作Wistar大鼠慢性点燃模型。在低温条件下迅速取脑,通过免疫组化检测皮质区NT-4的变化。结果实验性癫痫大鼠模型制作成功,精油组和癫痫模型组实验动物均达到点燃标准,与癫痫模型组动物相比,精油组大鼠潜伏期劈显延长,但持续时间之间的差别没有显著性。免疫组化检测实验结果表明：癫痫模型组大脑皮质NT--4含量比正常对照组增加,差异显著具有统计学意义性（P〈0．05）;精油组NT-4含量比癫痫模型组增加,差异显著有统计学意义（P〈0．01）。结论精油能够增强NT-4的表达,减轻癫痫发作时神经系统的损伤,可能有抗癫痫作用。     

颞叶癫痫大鼠学习记忆障碍与海马区PSD-95表达的关系

目的研究颞叶癫痫大鼠在Morris水迷宫中学习、记忆能力与大鼠海马区PSD-95表达变化的关系。方法随机将40只W istar大鼠分为海人酸（KA）组（28只）和对照组（12只）。KA组采用KA腹腔注射制作颞叶癫痫大鼠模型,根据是否出现自发性再发作（SRS）分SRS组（A组）、无SRS组（B组）;对照组（C组）注射生理盐水。通过Morris水迷宫测验观察各组大鼠注射KA或生理盐水2、6周时的空间学习、记忆能力,采用HE染色观察大鼠海马的组织病理学变化,免疫组化法检测大鼠海马CA1、CA3区PSD-95的表达。结果A组大鼠注射KA或生理盐水6周时海马区未见广泛神经元丢失及胶质增生,偶见局部神经元丢失及胶质增生;B、C组大鼠未见神经元丢失及胶质增生。与A组2周时及B、C组在2、6周时相比,A组6周时的学习、记忆能力明显下降（P〈0.01）,相应海马CA1、CA3区PSD-95表达均明显降低（P〈0.05）。结论颞叶癫痫长期反复发作时海马区PSD-95表达的减少,可能是导致颞叶癫痫大鼠学习、记忆障碍的机制之一。     

姜黄素对Aβ诱导的老年痴呆大鼠海马胶质纤维酸蛋白表达的影响

目的通过观察姜黄素对Aβ1～40诱导的阿尔茨海默病(AD)大鼠空间学习记忆能力及海马区星形胶质细胞胶质纤维酸蛋白(GFAP)表达的影响,探讨姜黄素的作用机制。方法选择健康SD大鼠48只,分为空白对照组、AD对照组、姜黄素给药组,每组16只。Aβ1～40诱导大鼠认知障碍模型,采用Morris水迷宫测定大鼠的学习记忆能力。采用免疫组织化学方法检测GFAP蛋白表达,RT-PCR检测GFAP mRNA表达。结果姜黄素干预后AD大鼠空间学习记忆能力明显改善(P<0.05);与空白对照组相比,AD对照组大鼠脑组织海马区可见星形胶质细胞GFAP表达明显增多(P<0.05),姜黄素给药组大鼠脑组织海马区GFAP mRNA及免疫阳性细胞表达较AD对照组下降(P<0.05)。结论姜黄素能够改善Aβ1～40诱导的AD模型大鼠空间学习记忆障碍,其机制可能与降低GFAP的表达和抑制星形胶质细胞活性有关。     

脑室内移植嗅成鞘细胞改善大鼠阿尔茨海默病的症状

目的探讨嗅成鞘细胞（olfactory ensheathing cells,OECs）脑室内注射对大鼠阿尔茨海默病的影响。方法sD大鼠双侧海马注射ABl．40,建立AD大鼠模型。实验动物分为四组：正常对照组、AD模型组、OECs移植组、MCSF注射组,每组10只。体外原代培养嗅成鞘细胞并将其移植至AD大鼠侧脑室。运用行为学测试、组织化学、原位杂交结合图像分析以及电镜等技术,观察、比较各组大鼠学习记忆能力、海马CA1区线粒体细胞色素氧化酶（cytochrome oxidase,COX）活性、细胞色素氧化酶Ⅱ型亚基（COⅡ）mRNA表达、一氧化氮合酶（nitric oxide synthase,NOS）阳性神经元数,以及CA1区神经元线粒体超微结构等指标的变化。结果与正常对照组比较,AD模型组大鼠空间学习记忆能力、海马CA1区线粒体COX活性及CO II mRNA表达、NOS阳性神经元数明显降低或减少俨〈0．05）,线粒体数肇减少,结构不清、肿胀、空泡样变、嵴断裂;嗅成鞘细胞移植明显上调上述指标俨〈0．05）,并对神经元线粒体超微结构有明显的保护作用。结论嗅成鞘细胞移植通过改善AD大鼠学习记忆能力、提高海马COX活性和CO1ImRNA、NOS阳性神经元表达以及保护神经元线粒体等作用,对大鼠阿尔茨海默病具有明显的治疗作用。     

低频电刺激小脑顶核对杏仁核点燃癫痫大鼠NMDAR及细胞内Ca^2＋的影响

目的研究低频电刺激（LFS）小脑顶核对杏仁核点燃大鼠癫痫模型海马N-甲基-D-天门冬氨酸受体（NMDAR）及细胞内Ca^2＋的影响。方法在成功点燃的大鼠小脑顶核埋置刺激电极,并对小脑顶核实施LFS,观察大鼠癫痫发作程度和持续时间,用免疫组化法测定海马NMDAR免疫阳性细胞计数及灰度值,流式细胞仪测定细胞内Ca^2＋浓度,并与正常对照组比较。结果与对照组比较,小脑顶核组大鼠发作级别降低、持续时间减少（P〈0．05）;刺激小脑顶核后海马NMDAR阳性细胞数明显减少、NMDAR阳性细胞灰度值升高（P〈0．05）;细胞内Ca^2＋浓度降低（P〈0．05）。结论LFS小脑顶核能有效抑制大鼠杏仁核点燃发作,其可能通过改变海马NMDAR水平及细胞内Ca^2＋浓度而发挥作用。     

杞蓟制剂对非酒精性脂肪性肝炎大鼠肝组织核转录因子-kB／IkB的影响

目的 观察非酒精性脂肪性肝炎（NASH）大鼠肝组织P65（NF-kB亚型）、核转录因子-kB抑制因子（IkB）-α（IkB亚型）蛋白和mRNA表达情况及杞蓟制剂对其影响。方法 采用高脂饮食复制大鼠NASH模型。实验分正常组对照、模型组、杞蓟制剂组、复方蛋氨酸胆碱片组。测定肝组织P65、IkB—α蛋白和mRNA的表达。结果与正常组相比，模型组大鼠肝组织P65、IkB—蛋白和mRNA表达明显增强（P〈0．01），且P65蛋白主要在胞核分布；与模型组相比，杞蓟制剂组大鼠肝组织P65、IkB—α蛋白和mRNA的表达明显降低（P〈0．05）。复方蛋氨酸胆碱片组大鼠肝组织P65、IkB-α蛋白和mRNA的表达虽有所降低，但与模型组相比差异无统计学意义（P〉0．05）。结论 NF—kB蛋白和mRNA表达增强及NF-kB蛋白活化增强参与了NASH的发病。杞蓟制剂能够抑制NF-kB（P65）蛋白和mRNA的表达，从而能防治NASH。     

洛哌丁胺对腹泻模型大鼠结肠水通道蛋白4表达的影响

目的检测腹泻模型大鼠在给予洛哌丁胺处理后结肠黏膜水通道蛋白4（AQP4）mRNA表达的变化,探讨洛哌丁胺在结肠水代谢巾的分子作用机制。方法采用逆转录聚合酶链反应（RT—PCR）分别对洛哌丁胺治疗组、模型对照组及正常对照组大鼠升、降结肠黏膜细胞AQP4mRNA表达进行半定量分析。结果①洛哌丁胺治疗组升、降结肠AQP4mRNA表达量均高于模型对照组及正常对照组（P〈0．01）。②模型对照组升结肠AQP4mRNA表达量较正常对照组大鼠减少（P〈0．05）,而降结肠表达差异无统计学意义（P〉0．05）。结论洛哌丁胺可以在转录水平上调腹泻大鼠结肠黏膜AQP4表达,使结肠对肠腔内水分的吸收增加。     

MMP-9及TIMP-1在支气管哮喘发病中作用及雷公藤多甙干预研究

目的观察支气管哮喘（简称哮喘）大鼠肺组织中细胞外基质（ECM）代谢相关因子基质金属蛋白酶9（MMP-9）及其抑制剂组织金属蛋白酶抑制剂1（TIMP-1）的表达,研究雷公藤多甙在哮喘肺组织ECM重塑中可能的作用及其机制。方法建立大鼠哮喘模型,采用逆转录-聚合酶链反应技术（RT-PCR）测定肺组织中MMP-9、TIMP-1mRNA的表达。结果哮喘组MMP-9、TIMP-1在肺组织中的蛋白表达明显高于对照组（P〈0．01）,应用药物雷公藤多甙干预后,MMP-9、TIMP-1蛋白表达明显低于哮喘组（P〈0．05）。哮喘组MMP-9、TIMP-1在肺组织的mRNA表达也高于对照组（P〈0．01）,应用雷公藤多甙药物干预后,MMP-9、TIMP．1在肺组织的mRNA表达明显低于哮喘组（P〈0．01）。哮喘组肺组织中MMP-9／TIMP-1〉1,明显高于对照组（P〈0．05）,应用药物雷公藤多甙干预后,MMP-9／TIMP．1〈1,明显低于对照组（P〈0．05）和哮喘组（P〈0．01）。结论雷公藤多甙可能下调MMP-9的表达,调节MMP-9／TIMP-1的平衡,干预细胞外基质重塑。     

用 ~(32)P~(45)Ca 双示踪方法研究大骨节病区粮喂养大白鼠钙磷代谢的变化

本文用~(32)P、~(45)Ca 双示踪技术研究大骨节病区粮喂养大白鼠的钙、磷代谢。结果表明:病区粮喂养动物的骨、软骨~(32)P、~(45)Ca 沉积减少,骨钙、磷含量显著增大,软骨磷脂、硫酸软骨素含量降低、胶元量增高。说明病区组动物发生了钙、磷代谢紊乱,有过早老化现象,并提出其改变的初步分析和可能有关的因素。病区粮加硒后,骨、软骨~(32)P 沉积率明显增高,软骨磷脂含量增加,骨磷量有所减少,而钙代谢无明显变化。表明硒与磷代谢有关。     

脑出血大鼠脑内移植神经干细胞后再生相关因子的表达

目的观察大鼠脑出血模型脑内移植神经干细胞(NSCs)后脑源性神经营养因子(BDNF)和血管内皮生长因子(VEGF)的表达及对血管生成的影响。方法 SD大鼠140只,随机分为假手术组(A组)、脑出血模型组(B组)、NSCs培养液移植组(C组),NSCs移植组(D组),每组35只。免疫组织化学方法观察各组术后6 h、1、4、7、14、21和28天BDNF、VEGF及CD34标记的微血管密度的变化,并进行相关分析。结果与A组比较,B组、C组BDNF的表达1天达高峰(P<0.05),D组表达高峰延迟至第4天,至28天仍高于A组(P<0.05);D组在所有时间点与B组比较,差异有统计学意义(P<0.05);D组在4天后的5个时间点与C组比较,差异有统计学意义(P<0.05)。B组、C组VEGF和微血管密度表达模式相同,均在1天达高峰;D组呈双高峰表达,在第4天和14天,至28天仍高于B组、C组的表达水平(P<0.05);相关分析显示,VEGF与微血管密度呈正相关(r=0.911,P<0.05)。结论脑出血大鼠脑内移植NSCs后,脑内BDNF及VEGF维持长时间的高表达,微血管密度增加。     

益气活血方对球囊损伤大鼠颈总动脉后PCNA表达的影响

目的 观察损伤血管细胞核增殖抗原(PCNA)的表达情况,探讨益气活血方对球囊损伤大鼠颈总动脉内膜平滑肌细胞(VSMCs)增殖的干预机制.方法 随机将108只雄性SD大鼠分为假手术组、模型组、阿司匹林组及益气活血方大、中、小剂量组.球囊损伤大鼠颈总动脉前给予高脂饲料喂养30 d,造成高脂血症;然后经右股动脉穿刺球囊扩张损伤大鼠左颈总动脉,造成颈总动脉管腔的狭窄.手术前1 d开始给药,每组动物于术后(5 d、14 d、28 d)处死,取损伤颈总动脉,常规固定、包埋、切片,分别做HE染色和免疫组织化学SABC法检测PCNA的表达.结果 各组PCNA均表达阳性,模型组明显增多,用益气活血方干预后14 d和28d PCNA表达量与模型组比较有统计学意义(P<0.05).结论 益气活血方对球囊损伤后动脉血管内膜VSMCs增生有明显的抑制作用,其机制可能与下调细胞核增殖抗原的表达,抑制血管内膜VSMCs的细胞分裂周期进程有关.     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对肺间质病变大鼠TGF-β1表达的影响

目的 观察重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR-Fc,益赛普)对博来霉索(BLM)-A5所致肺间质病变大鼠转化生长因子(TGF)-β1表达的影响.方法 SD雄性大鼠45只,随机分为对照组、模型组和rhTNFR-Fc治疗组(治疗组),每组15只.各组动物于第7、14、28天随机处死5只,取肺组织进行苏木素-伊红(HE)和Masson染色,用免疫组织化学法检测肺组织TGF-β1的表达水平.结果 对照组未见明显胶原沉积、炎症及纤维化改变,治疗组各时期肺泡炎和肺纤维化程度均较模型组轻,差异有统计学意义(P<0.01).模型组各时期TGF-β1的表达较对照组增加,差异有统计学意义(P<0.01);治疗组第7、14天TGF-β1的表达较对照组增加,差异有统计学意义(P<0.01),第28天稍高于对照组,但差异无统计学意义(P>0.05).治疗组第7天比模型组减低,差异有统计学意义(P<0.05),第14、28天比模型组减低差异有统计学意义(P<0.01).结论 rhTNFR-Fc能减轻BLM-A5诱导的大鼠肺泡炎和肺纤维化,可能与抑制肺组织TGF-β1表达有关.     

Memory T cells have gene expression patterns intermediate between naive and effector

The biological basis underlying differentiation of naive (NAI) T cells into effector (EFFE) and memory (MEM) cells is incompletely understood. Furthermore, whether NAI T cells serially differentiate into EFFE and then MEM cells (linear differentiation) or whether they concurrently differentiate into either EFFE or MEM cells (parallel differentiation) remains unresolved. We isolated NAI, EFFE, and MEM CD8(+) T cell subsets from human peripheral blood and analyzed their gene expression by using microarrays. We identified 156 genes that strongly differentiate NAI, EFFE, and MEM CD8(+) T cells; these genes provide previously unrecognized markers to help identify each cell type. Using several statistical approaches to analyze and group the data (standard heat-map and hierarchical clustering, a unique circular representation, multivariate analyses based on principal components, and a clustering method based on phylogenetic parsimony analysis), we assessed the lineage relationships between these subsets and showed that MEM cells have gene expression patterns intermediate between NAI and EFFE T cells. Our analysis suggests a common differentiation pathway to an intermediate state followed by a split into EFFE or MEM cells, hence supporting the parallel differentiation model. As such, conditions under which NAI T cells are activated may determine the magnitude of both EFFE and MEM cells, which arise subsequently. A better understanding of these conditions may be very useful in the design of future vaccine strategies to maximize MEM cell generation.     

The functional role of the carbohydrate in human melanoma associated antigens. I. Antigen-antibody complex formation

Messenger ribonucleic acids (mRNA) for tumor associated antigens (TAA) were isolated from human melanotic melanoma (HMMC-ShA, HMMC-WJP) and amelanotic melanoma (HMMC-Sr, HMMC-KM) cells. The melanoma cells were cultured in standard Eagle's MEM and in MEM supplemented with tunicamycin (Tu). The mRNAs were translated in vitro using wheat germ system. Using a standard immunodiffusion system, TAA from melanoma cells harvested from standard MEM differed from the TAA obtained from cells harvested from MEM supplemented with Tu, whereas, TAA pretreated with endo-beta-N-acetylglucosaminidase resembled TAA of cells harvested from MEM supplemented with Tu. The translation products synthesized by mRNA of melanotic melanoma cells resembled TAA extracted directly with 3 M KCl from the cells, but differed from TAA isolated directly from the cells or synthesized by mRNA from amelanotic melanoma cells. Therefore, the melanotic melanoma differed from the amelanotic melanoma cells in their genetic expression. The TAA-immunological reactivities are modulated by a post-translation step regulating genetic expression.     

Effects of some synthetic kynurenines on brain amino acids and nitric oxide after pentylenetetrazole administration to rats

We have previously proven that some synthetic kynurenines behave as antagonists of the N-methyl-d-aspartate receptor inhibiting neuronal subtype of nitric oxide synthase activity. We now investigate the anticonvulsant activity of four of these kynurenines in pentylenetetrazole (PTZ)-treated rats. The rats were treated with each kynurenine (10-160 mg/kg, s.c.) 30 min before PTZ administration (100 mg/kg, s.c.). Then, latency, duration and intensity of the first seizure and the percent animal survival were noted. PTZ-induced death was counteracted by high doses of kynurenines. Latency of the first seizure was significantly increased and its intensity reduced at the same doses, whereas the duration of the first seizure significantly decreased with doses of 20 mg/kg in most of the kynurenines tested. Three hours after PTZ administration, the surviving animals were sacrificed and the levels of brain amino acids and nitrite were measured. PTZ administration increased glutamate, glutamine, serine and taurine levels in different brain areas. High doses of kynurenines generally counteracted the effects of PTZ on excitatory amino acids, but they also reduced inhibitory aminoacids. However, the most consistent effect of kynurenines was the dose-dependent reduction of brain nitrite levels induced by PTZ. These results reveal a new family of anticonvulsant drugs that affect mainly to nitric oxide production in the brain.     

Effect of Pentylenetetrazole on Ethanol Intake, Ethanol Kinetics, and Social Behavior in Male Wistar Rats

Stress and anxiety are often implicated in excessive alcohol use. The nature of this interaction, however, is not understood. The aim of this study was to examine the effect of the anxiogenic agent, pentylenetetrazole (PTZ), on the acquisition and maintenance of ethanol drinking behavior in male Wistar rats. In rats maintained on a limited access procedure, with a choice between a 12% w/v ethanol (ETOH) solution and water available for 30 min each day, acute PTZ administration (1.5 to 15.0 mg/kg) did not modify ETOH intake. Chronic PTZ administration elicited a significant suppression in ETOH intake; however, this effect developed gradually over time. During the acquisition phase, chronic PTZ treatment also suppressed ETOH consumption. Chronic, but not acute, treatment with PTZ seemed to enhance water consumption. To assess whether the effect of PTZ on ETOH intake was due to either alterations in ETOH kinetics or behavior, blood ETOH levels and social interaction behaviour were examined. PTZ (15.0 mg/kg) produced a significant suppression in social interaction behavior, although tolerance developed to this effect on chronic PTZ administration. Both acute and chronic PTZ treatment (15 mg/kg) resulted in lower blood ETOH levels achieved after administration of 1.0 g/kg po of ETOH. Because the anxiogenic effect of PTZ was not maintained on repeated administration, yet the suppression of ETOH intake was only observed after chronic treatment, this suggests a dissociation between the processes regulating these behaviors.