高频电刺激大鼠丘脑底核抑制背侧中缝核5-羟色胺表达

本研究探讨高频电刺激丘脑底核对大鼠背侧中缝核5-羟色胺(5-HT)表达的影响。实验动物分两组,刺激组给予高频电流(130Hz,100μA,60μs)刺激大鼠右侧丘脑底核,对照组大鼠右侧丘脑底核植入电极,但无电流输出。刺激结束后,用免疫组织化学方法染色背侧中缝核5-HT能神经元,检测背侧中缝核5-HT能神经元的数量和平均灰度值。结果显示电刺激组背侧中缝核5-HT阳性神经元数目与对照组比明显减少(t(13)=3.786,P=0.002),并且神经递质5-HT表达量减少,平均灰度值显著增高(t(13)=7.917,P<0.001)。本实验结果表明高频电刺激丘脑底核对背侧中缝核5-HT能神经元有抑制作用,在应用高频电刺激丘脑底核治疗Parkison病运动障碍时出现的情绪障碍可能与其有关。     

5-羟色胺转运体在酗酒人脑干头侧中缝核群的表达

目的:观察酗酒人脑标本脑干头侧中缝核群5-羟色胺转运体(SERT)的表达变化。方法:应用免疫放射自显影的方法,显示SERT免疫反应强度在酗酒人脑标本脑干头侧中缝核群的变化,并与健康人脑标本比较。结果:健康组SERT免疫反应强标记信号在脑桥头侧和中脑主要集中分布在与中缝核群相同的区域。在酗酒人脑标本,正常分布于脑干头侧中缝核群的SERT免疫反应标记信号减弱;中缝正中核、中缝背核尾侧部、中缝背核束间部、中缝背核腹侧部、中缝背核背侧部的SERT免疫反应含量强度与健康组相应区域比较显著降低。结论:酗酒者头侧中缝核群SERT蛋白表达降低。     

延脑中缝核注射5-羟色胺对清醒大鼠胃运动的影响

中枢5-羟色胺(5-hydroxytryptamine,5-HT)作为一种神经递质或调节物集中在脑干中缝核群,外周5-HT主要位于胃肠道。以往研究表明:脑内和外周5-HT对饮食行为和消化道功能有着重要的调节作用。本研究采用应力传感器缝在胃窦浆膜面上记录胃运动的技术,观察向大鼠延脑中缝核注射5-HT和腹腔注射对氯苯丙氨酸(Para-chloropheny-     

大鼠脑干5-羟色胺神经元的分布——免疫细胞化学研究和定量分析

应用免疫细胞化学卵白素-生物素-过氧化物酶复合体(Avidin-Biotin-peroxidase Complex, ABC)法研究了大鼠脑干含5-羟色胺(5-Hydroxytryptamine,5-HT)的神经元的分布。证实5-HT神经元广泛分布于中缝核群和网状结构。在延髓表面弓状细胞群、极后区、蓝斑复合体和脚间核中也含5-HT神经元。定量分析表明:①5-HT神经元在脑干各部的比例约为:中脑61％,延髓31％,脑桥8％;②中缝核群内5-HT神经元约占脑干5-HT细胞总数的60％,中缝以外约占40％;③5-HT细胞在各核的分布比例为:中缝苍白核6.15％,中缝隐核5.77％,中缝大核5.15％,中缝桥核2.09％,中央上核3.44％,中缝背核35.82％,线形头侧核1.76％,延髓网状结构13.99％,脑桥网状结构和第Ⅳ脑室室底灰质5.40％,中脑网状结构20.42％。本文对中缝各核的细胞密度和5-HT细胞密度进行了抽样分析,结果表明:含5-HT细胞较高的核团有:中缝苍白核56.36％,中缝隐核48.40％,中缝背核44.10％;其次为中央上核33.38％,中缝大核25.77％;线形头侧核和中缝桥含5-HT细胞较低,分别为18.19％和17.37％。     

大鼠运动核内5-羟色胺1A、2A、5A受体的定位分布

为了阐明５ 羟色胺在中枢神经系统内与运动神经元结合的精确部位,本研究用免疫细胞化学技术分别观察了大鼠躯体运动核和内脏运动核内５ 羟色胺１Ａ、２Ａ、５Ａ 受体的定位分布。在躯体运动核内观察到：（１）５ 羟色胺１Ａ 受体样阳性神经元和纤维主要分布于动眼神经核、滑车神经核、三叉神经运动核、面神经核、舌下神经核和脊髓前角;（２）５ 羟色胺２Ａ 受体样阳性神经元主要见于动眼神经核、三叉神经运动核、面神经核、舌下神经核和脊髓前角,但阳性纤维和终末却密集地分布于三叉神经运动核、面神经核、舌下神经核和脊髓前角等处,除此之外动眼神经核、滑车神经核、展神经核和疑核内也能见到中等密度的阳性纤维和终末,纤维和终末的分布范围和染色浓度、密度都较神经元为明显;（３）少量淡染的５ 羟色胺５Ａ 受体样阳性神经元和稀疏的阳性纤维及终末主要见于三叉神经运动核、面神经核、舌下神经核和脊髓前角。在内脏运动核内观察的结果是：（１）动眼神经副交感核（Ｅ Ｗ 核）、上涎核、迷走神经背核、骶髓副交感核和胸髓侧角内仅有少量５ 羟色胺１Ａ 受体样阳性神经元、纤维和终末分布;（２）５ 羟色胺２Ａ 受体样阳性神经元和较密集的阳性纤维和终末见于Ｅ Ｗ 核、迷走神经背核、骶?     

大鼠臂旁核内5-羟色胺阳性纤维和终末的来源

本研究采用荧光金(FG)逆行追踪与5-羟色胺(5-HT)免疫荧光组化染色相结合的双重技术观察了臂旁核(PBN)内5-HT阳性神经纤维和终末的来源。将FG注入PBN后,FG逆标神经元主要分布在三叉神经核簇、脑干网状结构外侧小细胞部、中缝核簇和中脑导水管周围灰质(PAG);免疫荧光组化染色的结果显示5-HT阳性神经元主要位于中缝核簇和PAG;在中缝核簇和PAG内可见FG逆标并呈5-HT阳性的双重标记神经元。上述结果表明,中缝核簇和PAG内的5-HT能神经元向PBN发出投射,它们在躯体和内脏感觉信息的传递和调控方面发挥重要作用。     

5-羟色胺对断奶仔鼠应激腹泻的影响

目的探讨断奶仔鼠应激性腹泻发生与5-羟色胺(5-HT)的相关性及断奶仔鼠应激性腹泻的发生机制。方法将72只雄性21 d断奶ICR小鼠随机分为6组,分别为正常对照组、氢溴酸西酞普兰(CH)对照组、对氯苯丙氨酸(PCPA)对照组、应激腹泻组、CH+应激腹泻组及PCPA+应激腹泻组。腹腔注射给药组(CH 10 mg/kg或PCPA 300mg/kg)处理4h后进行应激腹泻处理,即给予番泻叶(0.4 kg/L)按15ml/kg体重灌胃+后肢束缚应激处理(持续4h),其对照组分别给予相同剂量的生理盐水。处理第5天,检测血糖值、小鼠血浆皮质酮(Cort)、5-HT含量和分布的变化。结果与对照组相比,应激腹泻组和CH对照组小鼠血浆和肠道5-HT含量、血糖值和血浆Cort含量升高,腹泻评分增加,但是增重显著减小;CH+应激腹泻组较应激腹泻组相比5-HT含量、血糖值和Cort含量和腹泻评分显著增加,增重显著减小。PCPA对照组5-HT含量显著降低,与正常对照组相比除血浆Cort含量增加以外无显著变化;PCPA+应激腹泻组与应激腹泻小鼠相比肠道5-HT含量显著降低,增重升高,血糖值降低,但未达到正常水平。结论断奶仔鼠应激性腹泻伴随5-HT增加,CH引起的肠道5-HT含量升高直接导致并加重断奶仔鼠应激性腹泻,PCPA诱导的5-HT含量降低可减弱小鼠腹泻程度。     

衰老大鼠脑干中缝核群5-羟色胺样神经元的变化——免疫金银染色法和图像定量分析研究

本文结合免疫金银染色(IGSS)法和图像定量分析,观察了幼年(2个月)、中年(10个月)和老年(24个月)Wistar大鼠脑于中缝核群5-羟色胺样(5-HT-LI)神经元的衰老变化。结果表明:(1)在衰老进程中,5-HT-LI神经元仅于老年组的中缝桥核和嘴侧线形核有显著减少(p<0.01);(2)从幼年到中年,5-HT-LI神经元截面积几乎增大一倍,但到老年无显著性变化;(3)5-HT-LI神经元的灰度值在整个衰老进程中呈降低趋势,提示神经元的5-羟色胺的含量和活性随增龄而升高;(4)5-HT-LI神经元形态的衰老变化表现为胞体萎缩,边缘皱折而呈不规则状,突起缩短锐化。     

多重脑震荡大鼠脑干中缝核团内5-羟色胺免疫阳性表达变化的研究

为了探讨多重脑震荡(multiple cerebral concussion,MCC)后大鼠中缝核团内5-羟色胺(5-HT)能神经元的变化规律,本实验采用自制单摆式机械打击装置复制MCC大鼠模型,研究伤后大鼠脑干中缝核团内5-HT及5-HT合成过程中的限速酶-色胺酸羟化酶(TPH)的表达。将56只大鼠随机分为7组:对照组、伤后1、2、4、8、16和24d组(n=8)。用免疫组织化学染色技术及图像分析法定量分析伤后大鼠脑干中缝核团内5-HT和TPH的表达变化。结果显示:(1)TPH免疫反应阳性产物在中缝背核、正中中缝核的表达在伤后2d时达到高峰,与正常对照组相比有显著性差异(P<0.05);中缝大核和中缝苍白核分别以伤后1d组和4d组阳性反应最强;(2)5-HT免疫反应阳性产物在中缝背核、正中中缝核的表达也在伤后2d时达到高峰,16、24d组基本恢复至正常水平;而中缝大核和中缝苍白核内5-HT的免疫反应性在各损伤组与正常对照组之间均无显著性差异(P>0.05)。以上结果表明,多重脑震荡后中缝核团内TPH和5-HT的表达增高,这为研究5-HT对MCC后认知障碍的影响提供了形态学依据。     

大鼠脑干至脊髓5-羟色胺投射纤维的起源——免疫细胞化学和酶组织化学双重标记法

本文应用 HRP 逆行追踪与免疫细胞化学相结合的双重标记技术,研究了大鼠脑干至脊髓5-羟色胺(5-HT)投射纤维的起源,并对中缝核群和网状结构内5-HT 阳性细胞、HRP 标记细胞和5-HT-HRP 双重标记细胞进行了数量分析。结果表明,1.在延髓、脑桥尾侧部,5-HT-HRP双重标记细胞约占5-HT 阳性细胞的55%;占 HRP 标记细胞的45%。中脑仅有极少数5-HT-HRP双重标记细胞。2.脑干至脊髓5-HT 下行投射,主要起源于中缝苍白核、中缝隐核、中缝大核和延髓浅表弓状纤维区,所含双重标记细胞约占双重标记细胞总数的68%。3.浅表弓状纤维区、舌下神经根间核、中缝苍白核和中缝隐核主要为5-HT 下行投射;中缝大核及周围网状结构主要为非5-HT 下行投射。     

反相高效液相色谱法测定血清中的5-羟色胺

建立了一种简便、快速测定血清中5-羟色胺的反相高效液相分析方法。采用的色谱柱为Bondapak C18不锈钢柱(5μm,250mm×4.6mmi.d.),流动相为:甲醇—水—乙酸(70:30:0.1 V/V),流速:1ml/min,荧光监测器,激发波长278nm,发射波长333 nm。结果表明:该方法线性关系良好(r=0.99992),5-羟色胺测定的线性范围为0.10~2.00μg/mL,最低检测限为0.02μg/mL(S/N=3)。5-羟色胺在血清样品中的加标回收率为92.37%~100.12%。该测定方法结果准确,可用于临床研究。     

Polymorphism C in the serotonin transporter gene (SLC6A4) in questionable dementia and Alzheimer's disease

The promoter region of the serotonin transporter gene (SLC6A4) shows a 22-bp tandem repeat polymorphism, indicated as polymorphism C, that has been associated to depression, obsessive-compulsive disorder, memory impairment, and anxiety. Less clear are data regarding its association with Alzheimer's disease (AD). No data were reported regarding its association with questionable dementia (QD). In this study we investigate for polymorphism C in the SLC6A4 gene 302 elderly subjects with a clinical diagnosis of AD (n = 105), QD (n = 88) and no cognitive impairment (n = 114) attending a geriatric ward. A community-dwelling sample of 390 healthy subjects was also included in the analysis. A significant higher prevalence of the C16/C16 genotype in AD than in QD was observed (37.14% vs. 3%; p = 0.041, OR 2.001, 95%CI 1.018–4.024), while no differences in the C16/C14 and C14/C14 genotypes as well as in the estimated allele frequencies were found. No further differences among the three groups of subjects were found, also when they were compared with the community-dwelling sample. These findings suggest that SLC6A4 gene variation may have only a minor role, if any, in AD or QD.     

Urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA) in the rat after immobilization stress

An increase of 100% in urinary 5-hydroxyindoleacetic acid, as a measure of release and catabolism of serotonin, is associated with the stress of 18 hr of restraint. Seventy percent of the rats developed gastric ulcers. Increased urinary 5-hydroxyindoleacetic acid levels were not observed in rats deprived of food for 42 hr.     

基于补体C5a在炎症性肠病中的作用研究

目的探索2,4,6-三硝基苯磺酸(TNBS)诱导炎症性肠病(IBD)发病机制中补体C5a的作用。方法选择SPF级雄性Sprague-Dawley(SD)大鼠60只,鼠龄6~8周,体质量180~220 g。随机分为3组,即空白对照组、TNBS IBD组、TNBS+抗C5a单克隆抗体干预组(简称治疗组)。IBD大鼠建模方法采用TNBS联合乙醇诱导灌肠法。治疗组在建模后立即每只腹腔注射抗C5a单克隆抗体1.5 mg干预。利用酶联免疫吸附分析(ELISA)法检测不同时间段大鼠血清中炎性介质C5a和白细胞介素(IL)-6、肿瘤坏死因子α(TNF-α)的表达水平;另给予C5a拮抗剂后,再次检测大鼠血清中IL-6、TNF-α的表达水平。结果C5a、IL-6、TNF-α伴随着炎症的进展而增高,伴随炎症的转归出现下降趋势,即血清中C5a、IL-6、TNF-α的变化趋势与炎症反应程度呈正向相关性;给予C5a拮抗剂后,TNBS IBD组与治疗组相比,IL-6、TNF-α的表达水平有明显下调趋势,差异有统计学意义(P<0.05)。结论初步判断抗C5a单克隆抗体可能通过抑制IL-6、TNF-α的表达,有效地降低了IBD炎症反应,提示C5a在IBD发病机制中发挥着重要作用。     

Effect of the C-terminal fragment of substance (SP5–11) on neuronal activity of the dorsal nucleus raphe

Laboratory of General Pathology of the Nervous System and Laboratory of Pathophysiology of Pain, Research Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 12, pp. 578–580, December, 1991.     

内皮舒张因子对大鼠颈动脉血栓形成的影响

在ＦｅＣｌ３损伤血管内皮导致的大鼠颈动脉血栓形成模型上发现血栓段血管组织环磷鸟苷（ｃＧＭＰ）含量减少，其内皮依赖性的乙酰胆碱（Ａｃｈ）扩血管作用减弱，而对非内皮依赖性的硝普钠扩血管反应无明显改变，病理形态观察可见动脉血栓形成和血管内皮严重损伤。用一氧化氮合成酶抑制剂ＬＮＮＡ处理后的动物明显地促进血栓形成，该作用可被一氧化氮前体Ｌ－精氨酸（Ｌ－Ａｒｇ）所拮抗，用Ｌ－Ａｒｇ处理后的动物显著减轻血栓形成     

Interstitial deletion of chromosome 5 in a neonate due to maternal insertion,ins(8;5)(p23;q33q35)

We describe an infant girl with an interstitial deletion of chromosome bands 5q33 to 5q35 inherited from a maternal interchromosomal insertion ins(8;5)(p23;q33q35) which was demonstrated by fluorescent in situ hybridization with whole chromosome paints. Physical anomalies included hypertonicity, microcephaly, short neck, apparently low-set ears, micrognathia, camptodactyly, mild rocker bottom feet, and hammer toe. Cardiac anomalies included a large ventricular septal defect, patent ductus arteriosus, pulmonary hypertension and hypoplastic right ventricle. She died at age 3 months. Am. J. Med. Genet. 86:289–293, 1999. © 1999 Wiley-Liss, Inc.     

Effect of the serotonin agonist buspirone on behaviour and hypothalamic-pituitary-adrenal axis in confident and fearful mink

Behavioural and hypothalamic-pituitary-adrenal (HPA) axis responses were investigated in farm mink (Mustela vison) selected for either confident or fearful behaviour for nine generations.Two groups of 2-year-old confident (n=12) and fearful (n=12) female mink were given the serotonin (5-HT) 1A receptor agonist buspirone (1.25 mg/kg/day), whereas two other groups of 2-year-old confident (n=12) and fearful (n=12) female mink were given saline, continuously for 5 weeks via osmotic minipumps. Behavioural reactions towards a novel object and towards humans were tested after 19-25 days, and HPA axis reactivity [adrenocorticotropic hormone (ACTH), cortisol] was measured after 28-31 days of treatment. Confident mink were more exploratory than fearful mink towards humans and a novel object. Confident mink spent more time in contact with the object than did fearful mink during saline-but not during buspirone-treatment. buspirone increased approach-withdrawal conflict behaviour towards a object in fearful mink only. The chronic dose of buspirone did not reduce fear towards humans and did not affect latencies to reaction, number of contacts, number and duration of manipulations, and stereotypic behaviour in a Novel Object test. Different HPA axis responses have emerged between confident and fearful mink, together with a different degree of fear-related behaviour. Fearful mink have a higher cortisol combined with a lower ACTH secretion than confident mink in response to capture and blood sampling. The central serotonergic system may be involved, and even though the precise underlying mechanisms are presently unknown, treatment with a 5-HT(1A) receptor agonist reduces the difference between confident and fearful mink in HPA axis reactivity.     

Effect of 5-azacytidine (5-aza) on UCP2 expression in human liver and colon cancer cells

The function of the uncoupling protein 2 (UCP2) is different for each cancer cell. However, the mechanism of expression is still unclear. DNA methylation affects protein expression and is one factor that transforms normal cells into cancer cells. In this study, the hepatocellular carcinoma Hep3B and HepG2 cells and colorectal cancer HT-29 cells were treated with 5-azacytidine (5-aza), a DNA demethylation agent, to observe the modification of UCP2 expression and the methylation degree in the UCP2 promoter region. Promoter basal activity and degree of UCP2 expression were measured in Hep3B, HepG2, and HT-29 cells. In addition, methylation-specific PCR (MSP) was performed to investigate the degree of methylation in the UCP2 promoter region. The methylation region in the UCP2 promoter was confirmed based on bisulfite sequencing. In Hep3B cells in which UCP2 mRNA was not transcribed, the promoter basal activity was significantly higher than in HT-29 or HepG2 cells in which UCP2 mRNA was transcribed. Treatment with 5-aza increased UCP2 expression in Hep3B and HT-29 cells; however, the expression in HepG2 cells was unchanged. The UCP2 promoter in Hep3B cells has numerous methylated regions compared with HT-29 and HepG2 cells. The results of the present study revealed that inhibition of UCP2 expression in Hep3B cells was due to methylation of the promoter region. Investigating the mechanism that induces UCP2 expression in cancer cells is important to understand the function of UCP2, which could aid in cancer treatment.     

TS、DPD表达水平对5-FU疗效的预测价值

目的:探讨胸苷酸合成酶(TS)及二氢嘧啶脱氢酶(DPD)转录水平及其对5-氟脲嘧啶疗效的预测价值.方法:利用逆转录聚合酶链反应(RT-PCR)方法从胃肠癌细胞株中扩增胸苷酸合成酶及二氢嘧啶脱氢酶的DNA片段,并与内参照基因GAPDH进行比较.结果:对5-FU敏感的细胞株MKN45 TS和DPD的表达水平较低,而其他对5-FU不敏感的细胞株则无此现象.结论:TS及DPD mRNA水平对应用5-氟脲嘧啶治疗胃肠癌的疗效具有一定预测价值.