新西兰兔四氯化碳肝硬化模型的建立与黄曲霉毒素B1对该模型肝脏急性损伤作用的观察

发布时间：2018-01-14 04:01

本文关键词：新西兰兔四氯化碳肝硬化模型的建立与黄曲霉毒素B1对该模型肝脏急性损伤作用的观察　出处：《广西医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 背景我们发现黄曲霉毒素B1(Aflatoxin B1,AFB1)对肝脏的损伤作用有个体差异;肝脏的基础疾病可能影响AFB1的体内代谢过程及其毒性作用从而影响个体肿瘤发生的危险性。因此探讨病毒性肝炎患者中AFB1暴露所造成的肝损伤可能有助于阐明肝炎病毒与化学毒物协同致癌的作用机制。目的本课题拟通过四氯化碳化学损伤处理建立新西兰兔肝硬化模型,观察该肝硬化模型建立后的稳定性,并初步观察AFB1对该模型的急性损伤作用,为下阶段利用该动物模型进行原发性肝癌的发病机制研究打下基础。方法第一阶段,将雄性新西兰兔24只随机分为A、B、C、D组,分别给予安慰剂,低、中、高剂量的四氯化碳(Carbon Tetrachloride,CCl_4)观察其体重(body weight,BW)、总蛋白(total protein,TP)、白蛋白(albumin,Alb)、球蛋白(globulin,Glo)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、γ-谷氨酰转移酶(γ-glutamyltransferase,G-GT)、白细胞(white blood cell,WBC)、红细胞(red blood cell,RBC)、血红蛋白(hemoglobin,HGB)和血小板(platelet,PLT)的变化,并定期进行随机抽样肝活检,直到活检发现肝硬化时停止给药;第二阶段,继续观察8周,检测其BW、TP、Alb、Glo、ALT、AST、G-GT、WBC、RBC、HGB和PLT,第3、5、8周时进行抽样肝活检观察肝硬化是否逆转;第三阶段,在观察第8周末将存活的新西兰兔平均分为两组,一组给予0.5mg/kg剂量的AFB1处理,观察其对新西兰兔的肝功能和血常规指标的影响。结果第一阶段,在第2-3周,C组有3只新西兰兔死亡,D组有5只新西兰兔死亡,在第10周,病理活检显示,A组新西兰兔全为S0期,B组新西兰兔有3只为S4期,2只新西兰兔为S3期;第二阶段,在第3周,病理活检示,B组5只新西兰兔肝纤维化分期与第一阶段第10周比较有统计学差异(P＜0.05),肝纤维化程度均有不同程度的减轻;第三阶段,0.5mg/kgAFB1处理后两小时,新西兰兔的TP、Alb、Glo、ALT、AST、G-GT、WBC、RBC、HGB和PLT无明显改变(P＞0.05)。结论按0.3ml/kg剂量给予新西兰雄性白兔皮下注射橄榄油与CCl_4按1:1配成50%的CCl_4溶液,每3天一次,连续10周,可以建成肝硬化模型。用本方法建成的肝硬化模型不稳定,在撤药后至多3周均发生不同程度的逆转,其肝功能和血常规各项指标都恢复到给药前水平。因此要获得稳定的肝硬化模型必须持续给药。给予该模型动物腹腔内注射0.5mg/kg剂量的AFB1,两小时内检查肝功能和血常规无明显异常。
[Abstract]:Background We found that the effects of aflatoxin B1 (AFB1) on liver injury were different in individuals. The underlying diseases of the liver may affect the metabolic process and toxicity of AFB1 in vivo and thus affect the risk of individual tumorigenesis. Therefore, to explore the possibility of liver damage caused by AFB1 exposure in patients with viral hepatitis. It is helpful to clarify the mechanism of synergistic carcinogenesis between hepatitis virus and chemical poison. Objective to establish the liver cirrhosis model of New Zealand rabbits by chemical injury of carbon tetrachloride and to observe the stability of the model and the acute injury effect of AFB1 on the model. It will lay a foundation for the study of the pathogenesis of primary liver cancer by using the animal model in the next stage. Methods in the first stage, 24 male New Zealand rabbits were randomly divided into two groups. High dose carbon Tetrachloride CCL _ (4) was used to observe the weight of carbon tetrachloride (BW). Total protein (TP), Albumin (Alb), globulin (Glo.). Alanine aminotransferase (alt). Aspartate aminotransferase (AST). 纬 -glutamyltransferase G-GTN (white blood cell). The changes of red blood cell blood, hemoglobin (HGB) and platelet platelet (PLT) were observed. Random liver biopsy was carried out regularly until the liver cirrhosis was detected by biopsy. In the second stage, eight weeks of observation were carried out to detect its BWN TPN, Alb, Glot, ASTG-GTN, WBC, RBCU, HGB and PLT, No. 3, 5. Liver biopsy was performed at 8 weeks to observe whether liver cirrhosis was reversed or not. In the third stage, the surviving New Zealand rabbits were divided into two groups at the end of the 8th week. One group was treated with 0.5 mg / kg AFB1. To observe its effect on liver function and blood routine indexes of New Zealand rabbits. Results in the first stage, 3 New Zealand rabbits died in group C at 2-3 weeks and 5 New Zealand rabbits in group D died. At the 10th week, pathological biopsy showed that all New Zealand rabbits in group A were S0. In group B, there were 3 New Zealand rabbits with stage S4 and 2 New Zealand rabbits with stage S3. In the second stage, at the third week, pathological biopsy showed that there was a significant difference in hepatic fibrosis stage between group B and the first 10 weeks (P < 0.05). The degree of hepatic fibrosis was alleviated in different degree. Two hours after the treatment of 0.5 mg / kg AFB1 in the third stage, New Zealand rabbits were treated with TTP Alb, Glob, GGTG, WBC1 and RBC. There was no significant change in HGB and PLT (P > 0.05). Conclusion New Zealand male rabbits were subcutaneously injected with olive oil at a dose of 0.3 ml / kg to form 50% CCl_4 solution with CCl_4 at 1: 1, once every 3 days for 10 weeks. The liver cirrhosis model established by this method is unstable and reversed in varying degrees at most 3 weeks after withdrawal. The liver function and blood routine were all recovered to the level before administration, so the stable liver cirrhosis model must be continuously administered. The model animal was given intraperitoneal injection of 0.5 mg / kg AFB1. There was no obvious abnormality in liver function and blood routine examination within two hours.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R575.2;R-332

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