无菌兔模型建立的规范化以及不同级别新西兰兔生物学特性的比较

发布时间：2018-05-26 09:19

  本文选题：无菌兔 + 剖宫产　； 参考：《中国协和医科大学》2008年硕士论文

【摘要】： 前言无菌兔模型没有寄生病原体,受到抗原刺激时,免疫反应强烈且单一,因此在生命科学研究中有着广阔的应用前景。但是,目前国外对无菌兔的研究多集中在寄生病原菌致病性机理方面,国内对无菌兔的研究还仅限于培养方法的优化,对无菌兔的研究还比较零散,没有对无菌兔的生物学特性进行系统研究。 目的随着生命科学的迅速发展,对实验动物质量的要求越来越高。本土化无菌动物的大规模推广和应用,其基础是本土化高等级实验动物模型建立方法的规范化以及其基础生物学数据测定。本课题正是针对这一趋势,对无菌兔模型进行了系统的研究,为无菌兔实验动物模型的推广和使用提供参考。 方法首先采用剖宫产的方法对新西兰兔进行剖腹净化,在无菌隔离器内剥离子宫取仔,用人工哺乳方法将仔兔培育成无菌兔。将部分离乳无菌兔接种4种正常寄生菌使之SPF化。4月龄时,分别对三组动物颈动脉采血,自动生化分析仪测定无菌兔、SPF兔和普通级兔血液学参数。颈动脉放血致死,从膀胱吸取尿液,测定尿常规,摘取动物各脏器,测量其长度和重量,计算脏器系数,各脏器进行病理切片,比较三组动物血常规、血生化、尿常规、脏器系数和组织学有无差异。 结果无菌兔模型建立过程中,共剖出仔兔31只,成活21只,成活率67.7%,获得了较高的成活率。无菌兔、SPF兔和普通兔生物学特性比较显示,三组动物血常规、血生化、尿常规、脏器系数和组织学多项参数差异显著: 1.血常规参数比较中,白细胞数、红细胞数、血红蛋白、红细胞容积、平均红细胞体积、平均血红蛋白含量、平均血红蛋白浓度、红细胞体积分部宽度、中性粒细胞百分比、单核细胞百分比、嗜酸性粒细胞百分比、嗜碱性粒细胞百分比、中性粒细胞数、淋巴细胞数、单核细胞数、嗜酸性粒细胞数、嗜碱性粒细胞数等17项指标上差异显著(P＜0.05),血小板数、平均血小板体积和淋巴细胞百分比差异不显著(P＞0.05); 2.血生化参数比较中,总蛋白、丙氨酸转氨酶、球蛋白、A/G、碱性磷酸酶、谷酰转肽酶、葡萄糖、肌酐、尿酸、Ca、高密度脂蛋白胆固醇等11项指标差异显著(P＜0.05),白蛋白、天冬氨酸转氨酶、尿素氮、磷、总胆固醇、甘油三酯、低密度脂蛋白胆固醇、磷酸肌酸激酶、乳酸脱氢酶9项指标差异不显著(P＞0.05); 3.尿常规参数比较中,尿比重、pH2项指标差异显著(P＜0.05),白细胞、亚硝酸盐、蛋白质、葡萄糖、酮体、尿胆元、胆红素、红细胞8项指标差异不显著(P＞0.05); 4.脏器系数比较中,左颌下腺、右颌下腺、心脏、肺、胆囊、脾脏、左肾上腺、右肾上腺、子宫+卵巢、左睾丸、胃长、蚓突长度、盲肠长度、盲肠总重、盲肠干重、胃总重、胃干重等17项指标差异显著(P＜0.05),脑、肝脏、左肾、右肾、胰脏、右睾丸、胃宽、小肠长度、直肠长度、大肠长度等10项指标差异不显著(P＞0.05); 5.组织学比较中,普通兔多例发生肝、肺、肾、脑、及肠道炎性病变;SPF兔、无菌兔与普通兔相比,未出现肝脏、肾、脑、肠道的炎性病变,间质性肺炎的比例下降。 结论通过规范的剖宫产手术和人工哺乳方法可以获得符合标准的实验无菌兔,具有可操作性。经过比较,无菌兔、SPF兔和普通兔生物学特性差异显著,不同微生物环境对新西兰兔的生物学特性有显著影响。
[Abstract]:There is no parasitic pathogen in the sterile rabbit model , and the immune response is strong and single , so it has a wide application prospect in the research of life sciences . However , the research on the sterile rabbit abroad is limited to the optimization of the culture method , and the research on the sterile rabbit is not scattered , and the biological characteristics of the sterile rabbit are not systematically studied .


Objective With the rapid development of life science , the requirement of experimental animal quality is getting higher and higher . The foundation of this method is the standardization of the method of establishing the animal model of high grade experiment and the measurement of its basic biological data . The research of this topic is the systematic research on the model of sterile rabbit , which provides reference for the promotion and use of the experimental animal model of sterile rabbit .


Methods First , caesarean section was used to purify New Zealand rabbits . In the aseptic isolator , the rabbits were peeled off and the rabbits were cultured into sterile rabbits . The blood parameters of the rabbits , SPF rabbits and common rabbits were determined by manual feeding method . The blood parameters of the rabbits , SPF rabbits and common rabbits were determined by using the artificial feeding method .


Results In the process of establishing a sterile rabbit model , 31 rabbits were dissected , 21 were survived , the survival rate was 67 . 7 % , and the survival rate was high . The biological characteristics of sterile rabbits , SPF rabbits and common rabbits showed that there were significant differences in blood routine , blood biochemistry , urine routine , organ coefficient and histological parameters in three groups :


1 . In the comparison of blood routine parameters , the number of white blood cells , the number of red blood cells , the contents of hemoglobin , the volume of red blood cells , the average red blood cell volume , the average hemoglobin content , the mean hemoglobin concentration , the percentage of neutrophil percentage , the number of neutrophils , the number of lymphocytes , the number of monocytes , the number of neutrophils , the number of neutrophils and the like were significantly different ( P & lt ; 0.05 ) , and the difference in platelet count , mean platelet volume and lymphocyte percentage was not significant ( P > 0.05 ) ;


2 . The indexes of total protein , alanine aminotransferase , globulin , A / G , alkaline phosphatase , glutamine transaminase , glucose , creatinine , uric acid , Ca , HDL - cholesterol were significantly different ( P < 0.05 ) , albumin , aspartate aminotransferase , urea nitrogen , phosphate , total cholesterol , triglyceride , low density lipoprotein cholesterol , creatine kinase and lactate dehydrogenase were not significant ( P > 0.05 ) .


3 . There was no significant difference between urine specific gravity and pH2 index ( P < 0.05 ) , white blood cell , nitrite , protein , glucose , ketone body , urinary bladder element , bilirubin and red blood cell index ( P > 0.05 ) .


4 . There were no significant differences in 17 indexes ( P > 0.05 ) , brain , liver , left kidney , right kidney , pancreas , right testis , stomach width , small intestine length , rectal length , large intestine length and so on .


5 . In the histological comparison , the common rabbits had liver , lung , kidney , brain , and intestinal inflammatory lesions ; SPF rabbits , sterile rabbits and common rabbits showed no liver , kidney , brain , intestinal inflammatory lesions , and the proportion of interstitial pneumonia decreased .


Conclusion The standard experimental sterile rabbits can be obtained by standard cesarean section and manual feeding method . The results showed that the difference of biological characteristics between the rabbits , SPF rabbits and common rabbits was significant , and the biological characteristics of New Zealand rabbits were significantly affected by different microbial environments .
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R-332

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