长期维持帕金森病食蟹猴模型的探索

利用MPTP制备的灵长类帕金森病（PD）模型,在症状及病理、生化改变方面均酷似人类PD,而且稳定可靠,对抗PD药物的反应（包括副作用）也同人类相似,这是目前所建立的最能反映人类PD特征的动物模型,因而被广泛用于PD发病机制、诊断及治疗方面的研究.但是大多数实验中,MPTP的使用方案都是急性处理,不能够模拟自然状态下PD渐进发展的过程,慢性的使用方案或许能克服这个局限,但是模型动物会逐渐出现进食困难、消化不良、便秘、腹泻等症状,进而出现营养不良、免疫功能低下、机会感染等并发症,动物本身的长期生存受到威胁,模型难以长期维持.因此,建立一套行之有效的模型维持方法就显得非常重要. 作者研究了自制营养乳对帕金森病食蟹猴模型的营养作用,并对帕金森病食蟹猴模型的长期维持方法进行了探索,报告如下。     

食蟹猴的实验室检查

非人灵长类 (ＮｏｎｈｕｍａｎＰｒｉｍａｔｅ)在解剖、生理和生化代谢等特点与人相似 ,在神经生理学、病理学、心理学及计划生育学等生物学和医学领域的研究中 ,常把猴作为实验动物来应用。以前使用最多的是弥猴属中的恒河猴 ,但近年来食蟹猴作为实验动物应用的数量日益增多 ,这是因为食蟹猴体形小 ,性情温顺 ,便于实验操作 ,更重要的是已积累了丰富的基础数据 (各项生理、病理指标及各种实验模型等 ) ,具备了作为实验动物的必要条件[1] 。食蟹猴与其它非人灵长类一样会感染各种病 ,有些疾病是人猴共患的 ,为保证实验用猴的质量和接触猴的…     

食蟹猴的养殖——越南猴场考察报告

食蟹猴的养殖——越南猴场考察报告徐汉涛（南京农业大学动物科技学院２１００９５１９９６年８月,笔者应邀对越南猴场进行为期４０天的实地考察和学术交流。越南猴场位于胡志明市以东约３０公里的龙城县境内,饲养食蟹猴５０００余只和少量恒河猴、平顶猴、红面猴等,至...     

2型糖尿病食蟹猴白细胞NF-κB和IκBα表达相关性分析

比较了食蟹猴2型糖尿病(T2DM)不同发病阶段与正常对照组的空腹血糖值(FPG)、甘油三酯(TG)、胆固醇(CHOL),低密度脂蛋白胆固醇(LDL-C),高密度脂蛋白胆固醇(HDL-C)在血液中含量,及荧光定量PCR方法测定的NF-κB和IκBα在外周血白细胞中表达量.结果显示,FPG、CHOL和LDL-C随疾病进行呈...     

食蟹猴肺炎球菌病的诊断与防治

广西某野生动物保护站下属养猴场发生一起食蟹猴其症状呈急性经过、死亡率高的疾病,经临床观察、病理剖检及实验室检验,确诊为食蟹猴肺炎球菌病。１发病情况该养猴场于１９９８年４月２３日凌晨,从中越边境长途运输引进一批野外捕获的食蟹猴共１２４只,在到场６个小时...     

浅谈食蟹猴阴茎红肿的治疗

1 发病情况 在本实验动物养殖基地的食蟹猴阴茎红肿的病例时有发现,全年均可发病.笔者在近四年的兽医临床中共收治此类病例近60例,通过治疗都能治愈,无一死亡,现将治疗方法介绍如下.     

实验室饲养食蟹猴的常见症状及护理

食蟹猴作为实验动物的使用越来越多,特别是在医学实验室中.在饲养和使用食蟹猴的过程中,有时会发现个体异常的食蟹猴,出现一些临床症状,这时就需要工作人员特别是兽医对症状做出及时准确的判断、分析原因,加强护理,积极保障实验室中动物的健康和正常使用,为动物实验提供可靠的动物,提升实验质量.     

氯胺酮麻醉对食蟹猴血压的影响研究

目的:观察使用氯胺酮麻醉对食蟹猴血压的影响状况。方法:10只食蟹猴先在清醒状态下测量血压,再使用盐酸氯胺酮注射液10 mg/kg肌肉注射全身麻醉后,测量其麻醉后血压,比较两种状态下血压之间的差异。结果:食蟹猴在使用氯胺酮麻醉后收缩压、舒张压和平均动脉压升高。结论:食蟹猴使用氯胺酮进行全身麻醉后,对血压产生明显影响。     

Effects of Momordica grosvenori Saponins on Type 2 Diabetes Rats Induced by High-sucrose/High-fat Diet and Streptozotocin

The purpose of the experiment was to study the effect of Momordica grosvenori saponins on improving the effect of high-sucrose/high-fat diet combined with streptozotocin induced type 2 diabetes in rats.The type 2 diabetes rat model was established with a high-sucrose and high-fat diet combined with streptozotocin.Successfully modeled rats were given the total saponins of Momordica grosvenori (100,200,400 mg/kg) or metformin (268 mg/kg) by gavage,once a day for 4 weeks.Recording the "three more and one less"index (24 h internal urine output,water intake and feed intake,body weight);Detecting of biochemical indicators,analysis of fasting blood glucose (FBG),serum total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),insulin (INS),catalase (CAT),superoxide dismutase (SOD),malondialdehyde (MDA),alanine aminotransferase (AST),aspartate aminotransferase (ALT),blood urea nitrogen (BUN) and creatinine (Cr) levels.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the pancreas.Compared with the model group,the feed intake,water intake,urine output,blood indicators FBG,INS,MDA,TC,TG,BUN and Cr levels,AST and ALT activities were significantly or extremely significantly decreased in the total saponins of Momordica grosvenori each dose group and metformin group (P<0.05;P<0.01),while CAT,SOD activities and HDL-C level were significantly or extremely significantly increased (P<0.05;P<0.01).In pancreatic tissue,the number of pancreatic cells was increased significantly,and the pathological damage of the pancreas was significantly reduced. Momordica grosvenori saponins and metformin had obvious effects on improving glucose metabolism disorders,and at the same time could significantly reduce insulin resistance,enhance the body’s ability to resist oxidative stress and improve liver and kidney functions.     

罗汉果总皂苷对高糖高脂饲料联合链脲佐菌素致2型糖尿病大鼠的作用

试验旨在研究罗汉果总皂苷改善高糖高脂饲料联合链脲佐菌素致2型糖尿病大鼠的作用,以高糖高脂饮食联合链脲佐菌素建立2型糖尿病大鼠模型,造模成功大鼠灌胃给予罗汉果总皂苷（100、200、400 mg/kg）或二甲双胍（268 mg/kg）,每天1次,连续给药4周后,记录"三多一少"指标（24 h内尿量、饮水量和采食量,体重）;检测生化指标,分析空腹血糖值（FBG）、血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、胰岛素（INS）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、丙二醛（MDA）、丙氨酸氨基转移酶（AST）、天冬氨酸氨基转移酶（ALT）、尿素氮（BUN）和肌酐（Cr）水平,采用苏木素-伊红染色法（HE）观察胰腺的病理学改变。与模型组比较,罗汉果总皂苷各剂量组及二甲双胍组均可显著或极显著降低采食量、饮水量、尿量,血液指标FBG、INS、MDA、TC、TG、BUN和Cr水平、以及AST和ALT活性（P<0.05;P<0.01）,而CAT、SOD活性和HDL-C水平显著或极显著升高（P<0.05;P<0.01）;胰腺组织中,胰腺细胞数目明显增多,胰腺病理性损伤明显减轻。罗汉果总皂苷及二甲双胍均具有明显改善糖代谢紊乱的作用,同时可明显减轻胰岛素抵抗,增强机体抗氧化应激的能力和改善肝肾功能。     

核转录因子NF-E2相关因子基因缺失对高脂饲粮诱导非酒精性脂肪肝模型小鼠肝脏的影响

本试验旨在研究敲除核转录因子NF-E2相关因子(Nrf2)对高脂饲粮诱导小鼠肝脏氧化应激的影响。选择雄性野生型(WT)和Nrf2基因敲除(KO)ICR小鼠各20只,分别随机分成2组,每组各10只。1组WT和1组KO小鼠饲喂普通饲粮,为对照组,另2组饲喂高脂饲粮,为高脂组。试验期8周。结果显示,1)与对照组相比,WT和KO高脂组小鼠血清总胆固醇(TC)、低密度脂蛋白(LDL)含量和谷丙转氨酶(ALT)、谷草转氨酶(AST)活性极显著升高(P0.01),碱性磷酸酶(ALP)活性显著或极显著升高(P0.05或P0.01),总蛋白(TP)含量极显著降低(P0.01),但甘油三酯(TG)和高密度脂蛋白(HDL)含量变化不显著(P0.05)。2)WT高脂组小鼠肝脏表现出小泡性脂肪变性,KO高脂组小鼠肝脏表现出严重的大泡性脂肪变性和温和的小泡性脂肪变性。3)与对照组相比,WT和KO高脂组小鼠肝脏中谷胱甘肽(GSH)含量和超氧化物歧化酶(SOD)活性显著或极显著降低(P0.05或P0.01),而过氧化物酶(POD)活性和丙二醛(MDA)含量显著或极显著升高(P0.05或P0.01)。与WT小鼠相比,KO高脂组小鼠肝脏中MDA含量增加49%,但GSH含量及SOD、POD活性变化不显著(P0.05)。与WT对照组小鼠相比,KO高脂组小鼠肝脏中SOD、POD活性和MDA含量显著或极显著升高(P0.05或P0.01),而GSH含量极显著降低(P0.01)。由此可见,高脂诱导Nrf2基因缺失小鼠发生较严重的脂肪变性,同时高脂饲粮诱导的非酒精性脂肪肝模型小鼠肝脏发生氧化应激。     

链脲佐菌素诱导小鼠Ⅱ型糖尿病模型的研究

通过注射链脲佐菌素(STZ)诱导建立与人类Ⅱ型糖尿病(NIDDM)相似的昆明鼠和C57鼠模型,比较两种鼠建模剂量和成模率的差异。将昆明鼠和C57小鼠随机分为正常对照组(A)及低、中、高剂量(每天50、70、90 mg/kg)用药组(B)(n=10)。正常对照组(A)饲喂普通饲料,用药组(B)饲喂高脂高糖饲料;第4周开始,A组腹腔注射柠檬酸缓冲液,B组腹腔注射STZ,连续4 d。测定给药前和给药后3、7、15 d空腹血糖和体重,观察小鼠饮食、饮水和排尿情况。给药前空腹血糖A、B组间无显著性差异(P0.05);昆明鼠用药3 d后中、高剂量组均升高并达到建模标准,C57鼠用药7 d后中、高剂量组达到建模标准。两种小鼠用药3 d后各组体重均减低,并与对照组间有极显著差异(P0.01)。C57鼠,STZ注射剂量越高,体重减轻幅度越大;并且自造模成功后,各用药组体重均出现逐日降低的趋势。诱导建立NIDDM模型的适宜STZ剂量昆明鼠为每天50 mg/kg和70 mg/kg,并以后者为佳;C57鼠为每天70 mg/kg。     

Development of a Delayed-Type Hypersensitivity (DTH) Model in the Cynomolgus Monkey

Although a T-dependent antibody response (TDAR) assay is generally recommended as the first-line immune function assay in nonclinical immunotoxicity evaluation, second-line assays such as delayed-type hypersensitivity (DTH) to measure cell-mediated responses can provide helpful additional information. In this study, male Cynomolgus monkeys were injected intramuscularly either once or twice with 1 mg Keyhole Limpet Hemocyanin (KLH) or twice with a commercially available tetanus vaccine (40 IU tetanus toxoid + 0.06 mg aluminum hydroxide). All animals were subsequently challenged by intradermal injections of the same antigen or aluminum hydroxide after 4, 6 and 8 weeks. Clinical reactions at the injection sites were scored 24, 48 and 72 h post challenge. Skin biopsies were taken on completion of the observation period after each challenge for standard histological examination and immunolabeling using CD3 (T lymphocytes), CD19 (B lymphocytes) and CD68 (macrophages) antibodies. Tetanus toxoid induced stronger clinical reactions than KLH, whereas aluminum hydroxide induced no clinical reaction. Perivascular mononuclear cell infiltrates, a histopathological finding consistent with a DTH reaction, were seen after all challenges with tetanus toxoid or KLH, but not with aluminum hydroxide. Immunohistochemistry evidenced the presence of T lymphocytes and macrophages within these infiltrates. These results suggest that tetanus toxoid adjuvanted with aluminum hydroxide can induce a consistent DTH response for use as a model of cell-mediated response in Cynomolgus monkeys.     

Histopathological Changes in the Pancreas from a Spontaneous Hyperglycemic Cynomolgus Monkey

Morphological and immunohistochemical examinations were carried out on the pancreas of a hyperglycemic 5-year-old male cynomolgus monkey. Body weight gradually decreased from 6 months before termination, accompanying a slight reduction in food consumption and anorexia for the last 2 days. The blood glucose level was markedly elevated when examined at termination. Histopathologically, in the exocrine pancreas, diffuse hyperplasia of centroacinar and intercalated duct cells and diffuse atrophy of acinar cells with sporadic apoptosis were observed, although most centroacinar and intercalated duct cells were proliferating cell nuclear antigen (PCNA)-positive in both the present case and age-matched control animals. In the endocrine pancreas, the islets tended to be hypertrophic, with an increase in insulin-positive cells in comparison with the age-matched control animals. PCNA-positive cells also tended to increase in the islets, although positive cells for phospho-histone H3, a marker for mitotic cells, were not detected in the endocrine and exocrine pancreas. Moreover, neither inflammation nor amyloidosis was noted in the islets. In conclusion, the present case probably suffered from early-stage type 2 diabetes mellitus, and it provides fundamental information concerning pancreatic histopathology under insulin-related derangement in monkeys.     

猪圆环病毒Ⅱ型体外诱导3D4/2细胞炎症模型的建立

【目的】通过对猪圆环病毒Ⅱ型（PCV2）体外感染3D4/2细胞浓度、时间与细胞炎症水平进行探讨,建立PCV2体外感染3D4/2细胞炎症反应模型,以期为后期药物调控PCV2诱发3D4/2细胞炎症反应的研究奠定基础。【方法】将3D4/2细胞分为对照组及10⁰、10^-1、10^-2和10^-3 PCV2感染组,每组3个重复。对照组用DMEM培养,各PCV2感染组用不同稀释倍数PCV2液培养,2 h后均更换为含5%胎牛血清（FBS）的DMEM维持液进行培养,培养4、8、12和24 h后分别收集细胞及细胞上清液。采用Griess法检测一氧化氮（NO）水平,DCFH-DA荧光探针法检测活性氧（ROS）水平,酶标法检测还原型谷胱甘肽（GSH）水平,分光光度法检测黄嘌呤氧化酶（XOD）和髓过氧化物酶（MPO）活性,ELISA法测定白细胞介素-1β（IL-1β）、IL-6、肿瘤坏死因子-α（TNF-α）、IL-10、γ干扰素（IFN-γ）、IL-8、单核细胞趋化蛋白1（MCP-1）以及环氧合酶1（COX-1）和COX-2的分泌水平。【结果】10⁰至10^-3 PCV2作用4、8、12和24 h均能够成功感染3D4/2细胞。与对照组相比,10⁰ PCV2在感染3D4/2细胞4、8、12、24 h后ROS水平均极显著升高（P<0.01）,10^-1至10^-3 PCV2感染3D4/2细胞8、12、24 h后ROS水平显著或极显著升高（P<0.05;P<0.01）;10⁰至10^-3 PCV2感染3D4/2细胞8、12、24 h后,细胞内NO浓度及MPO活性显著提高（P<0.05）,细胞上清液中的IL-1β、IL-6、TNF-α、IL-10、IFN-γ、IL-8和MCP-1水平及COX-1活性均显著或极显著升高（P<0.05;P<0.01）,其中10⁰ PCV2感染3D4/2细胞后,各炎症因子水平上升最显著,且随着时间的延长,NO浓度逐渐升高,XOD活性逐渐降低。【结论】PCV2可诱导3D4/2细胞炎症反应,且10⁰ PCV2体外感染3D4/2细胞4~12 h是建立炎症模型的最佳条件。     

高精料饲粮诱发瘤胃胀气机理的研究进展

生产中为了追求高产,反刍动物不可避免地需要饲喂高精料饲粮来满足其生产需要,而反刍动物饲喂高精料饲粮易诱发瘤胃胀气和瘤胃酸中毒,显著降低动物的生产性能,甚至还会导致动物死亡,给养殖业造成重大经济损失。早期研究认为瘤胃产气过多过快是高精料饲粮诱发瘤胃胀气的主要原因,但随着科技的进步,越来越多的研究表明,瘤胃气体被泡沫包裹无法正常排放才是高精料饲粮诱发瘤胃胀气的根本原因。瘤胃泡沫形成的确切机制还有待研究,来源于饲粮的、瘤胃微生物合成的以及瘤胃微生物酵解饲粮产生的蛋白质、多糖和羧酸盐可能充当了瘤胃泡沫的起泡剂或泡沫稳定剂。     

抗性淀粉对饮食诱导肥胖大鼠排便状况及肠道菌群的影响

本试验旨在探讨抗性淀粉(RS)对饮食诱导肥胖(DIO)大鼠排便状况及肠道菌群的影响,为合理利用RS提供依据。选用100只健康雄性SD大鼠,随机分为2组,对照组10只饲喂基础饲粮,高脂组(HF组)90只饲喂高脂饲粮,7周后根据体重从HF组筛选出DIO大鼠27只,随机分为3组,每组9只,分别为HF组(饲喂高脂饲粮)、高脂饲粮抗性淀粉组(HFRS组,饲喂含10%RS的高脂饲粮)、高脂饲粮抗性淀粉+硫酸葡聚糖(DS)组(HFRS+DS组,饲喂含10%RS的高脂饲粮),试验期间HFRS+DS组每天用5%DS 1 m L灌胃,其他组分别用蒸馏水1 m L灌胃,试验期5周。于第7、12周收集新鲜成形大便1 g,稀释后用选择性培养基进行肠球菌、肠杆菌、双歧杆菌、乳酸杆菌、类杆菌检测并计数菌落总数。结果表明:1)试验第7周时,与对照组相比,HF组大鼠粪便湿重、粪便含水率以及粪样中肠球菌、双歧杆菌、乳酸菌、类杆菌数量显著降低(P0.05),粪便颗粒数、粪样中肠杆菌数量显著升高(P0.05)。2)试验第12周时,与对照组相比,HF组DIO大鼠粪便湿重、粪便含水率以及粪样中肠球菌、双歧杆菌、乳酸菌、类杆菌数量显著降低(P0.05),粪便颗粒数、粪样中肠杆菌数量显著升高(P0.05);与HF组相比,HFRS组DIO大鼠粪便湿重、粪便干重、粪便含水率以及粪样中肠球菌、双歧杆菌、乳酸菌、类杆菌数量均显著升高(P0.05),粪便颗粒数、粪样中肠杆菌数量显著降低(P0.05);与HFRS组相比,HFRS+DS组DIO大鼠粪便湿重、粪便含水率以及粪样中双歧杆菌、乳酸菌、类杆菌数量显著降低(P0.05),粪样中肠杆菌、肠球菌数量显著增加(P0.05)。由此可见,本试验条件下,RS可改善DIO大鼠排便状况和高脂饲粮造成的肠道菌群紊乱。     

Effect of a Large Dose of Di (2-ethylhexyl) phthalate (DEHP) on Hepatic Peroxisome in Cynomolgus Monkeys (Macaca Fascicularis)

To elucidate the effect of a large dose of di (2-ethylhexyl) phthalate (DEHP), a plasticizer and peroxisome proliferator-activated receptor-α (PPARα) agonist, on hepatic peroxisomes, we orally administered 1,000 mg/kg/day, once daily, to 3 male and 4 female cynomolgus monkeys for 28 days consecutively. Light-microscopic and electron microscopic examinations of the liver were carried out in conjunction with measurement of the hepatic fatty acid β-oxidation system (FAOS), carnitine acetyltransferase (CAT) and carnitine palmitoyltransferase (CPT) activities, which are peroxisomal and/or mitochondrial enzyme activities. Electron microscopically, enlargement of the mitochondria was observed with lamellar orientation of the cristae along the major axis. Although the number of peroxisomes showed a tendency to increase when compared with those in a biopsied specimen before treatment, no abnormality in morphology was observed. A slight increase in CPT activity was noted at termination. No changes were noted in hepatic FAOS or CAT activity. In conclusion, although repeated oral treatment of cynomolgus monkeys with a large dose of DEHP induced a subtle increase in the numbers of peroxisomes with slight enlargements of the mitochondria, this low-sensitivity response to peroxisome proliferators in cynomolgus monkeys was considered to be closer to the response in humans than that in rodents.