本文選題：無菌兔 + 剖宮產(chǎn)　； 參考：《中國協(xié)和醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 前言無菌兔模型沒有寄生病原體,受到抗原刺激時(shí),免疫反應(yīng)強(qiáng)烈且單一,因此在生命科學(xué)研究中有著廣闊的應(yīng)用前景。但是,目前國外對(duì)無菌兔的研究多集中在寄生病原菌致病性機(jī)理方面,國內(nèi)對(duì)無菌兔的研究還僅限于培養(yǎng)方法的優(yōu)化,對(duì)無菌兔的研究還比較零散,沒有對(duì)無菌兔的生物學(xué)特性進(jìn)行系統(tǒng)研究。 目的隨著生命科學(xué)的迅速發(fā)展,對(duì)實(shí)驗(yàn)動(dòng)物質(zhì)量的要求越來越高。本土化無菌動(dòng)物的大規(guī)模推廣和應(yīng)用,其基礎(chǔ)是本土化高等級(jí)實(shí)驗(yàn)動(dòng)物模型建立方法的規(guī)范化以及其基礎(chǔ)生物學(xué)數(shù)據(jù)測(cè)定。本課題正是針對(duì)這一趨勢(shì),對(duì)無菌兔模型進(jìn)行了系統(tǒng)的研究,為無菌兔實(shí)驗(yàn)動(dòng)物模型的推廣和使用提供參考。 方法首先采用剖宮產(chǎn)的方法對(duì)新西蘭兔進(jìn)行剖腹凈化,在無菌隔離器內(nèi)剝離子宮取仔,用人工哺乳方法將仔兔培育成無菌兔。將部分離乳無菌兔接種4種正常寄生菌使之SPF化。4月齡時(shí),分別對(duì)三組動(dòng)物頸動(dòng)脈采血,自動(dòng)生化分析儀測(cè)定無菌兔、SPF兔和普通級(jí)兔血液學(xué)參數(shù)。頸動(dòng)脈放血致死,從膀胱吸取尿液,測(cè)定尿常規(guī),摘取動(dòng)物各臟器,測(cè)量其長度和重量,計(jì)算臟器系數(shù),各臟器進(jìn)行病理切片,比較三組動(dòng)物血常規(guī)、血生化、尿常規(guī)、臟器系數(shù)和組織學(xué)有無差異。 結(jié)果無菌兔模型建立過程中,共剖出仔兔31只,成活21只,成活率67.7%,獲得了較高的成活率。無菌兔、SPF兔和普通兔生物學(xué)特性比較顯示,三組動(dòng)物血常規(guī)、血生化、尿常規(guī)、臟器系數(shù)和組織學(xué)多項(xiàng)參數(shù)差異顯著: 1.血常規(guī)參數(shù)比較中,白細(xì)胞數(shù)、紅細(xì)胞數(shù)、血紅蛋白、紅細(xì)胞容積、平均紅細(xì)胞體積、平均血紅蛋白含量、平均血紅蛋白濃度、紅細(xì)胞體積分部寬度、中性粒細(xì)胞百分比、單核細(xì)胞百分比、嗜酸性粒細(xì)胞百分比、嗜堿性粒細(xì)胞百分比、中性粒細(xì)胞數(shù)、淋巴細(xì)胞數(shù)、單核細(xì)胞數(shù)、嗜酸性粒細(xì)胞數(shù)、嗜堿性粒細(xì)胞數(shù)等17項(xiàng)指標(biāo)上差異顯著(P＜0.05),血小板數(shù)、平均血小板體積和淋巴細(xì)胞百分比差異不顯著(P＞0.05); 2.血生化參數(shù)比較中,總蛋白、丙氨酸轉(zhuǎn)氨酶、球蛋白、A/G、堿性磷酸酶、谷酰轉(zhuǎn)肽酶、葡萄糖、肌酐、尿酸、Ca、高密度脂蛋白膽固醇等11項(xiàng)指標(biāo)差異顯著(P＜0.05),白蛋白、天冬氨酸轉(zhuǎn)氨酶、尿素氮、磷、總膽固醇、甘油三酯、低密度脂蛋白膽固醇、磷酸肌酸激酶、乳酸脫氫酶9項(xiàng)指標(biāo)差異不顯著(P＞0.05); 3.尿常規(guī)參數(shù)比較中,尿比重、pH2項(xiàng)指標(biāo)差異顯著(P＜0.05),白細(xì)胞、亞硝酸鹽、蛋白質(zhì)、葡萄糖、酮體、尿膽元、膽紅素、紅細(xì)胞8項(xiàng)指標(biāo)差異不顯著(P＞0.05); 4.臟器系數(shù)比較中,左頜下腺、右頜下腺、心臟、肺、膽囊、脾臟、左腎上腺、右腎上腺、子宮+卵巢、左睪丸、胃長、蚓突長度、盲腸長度、盲腸總重、盲腸干重、胃總重、胃干重等17項(xiàng)指標(biāo)差異顯著(P＜0.05),腦、肝臟、左腎、右腎、胰臟、右睪丸、胃寬、小腸長度、直腸長度、大腸長度等10項(xiàng)指標(biāo)差異不顯著(P＞0.05); 5.組織學(xué)比較中,普通兔多例發(fā)生肝、肺、腎、腦、及腸道炎性病變;SPF兔、無菌兔與普通兔相比,未出現(xiàn)肝臟、腎、腦、腸道的炎性病變,間質(zhì)性肺炎的比例下降。 結(jié)論通過規(guī)范的剖宮產(chǎn)手術(shù)和人工哺乳方法可以獲得符合標(biāo)準(zhǔn)的實(shí)驗(yàn)無菌兔,具有可操作性。經(jīng)過比較,無菌兔、SPF兔和普通兔生物學(xué)特性差異顯著,不同微生物環(huán)境對(duì)新西蘭兔的生物學(xué)特性有顯著影響。
[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 .
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R-332

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本文編號(hào)：1936693