C57-ras转基因小鼠模型的MNU验证实验  

Validation study of C57-ras transgenic mice using MNU

作　　者：吕建军[1,2] 刘甦苏[1,3] 左琴[1,3] 周舒雅[1,3] 杨艳伟[1,2] 张頔[1,2] 汪巨峰[1,2] 范昌发[1,3] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]国家药物安全评价监测中心,北京100176 [3]国家啮齿类实验动物种子中心,北京100079

出　　处：《药物分析杂志》2013年第11期1935-1941,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技重大专项(No.2008ZX09305)资助

摘　　要：目的:建立国内用于药物临床前安全评价致癌性实验替代方法的C57-ras转基因小鼠模型,并通过甲基亚硝基脲(Nmethyl-N-nitrosourea,MNU)进行初步验证。方法:C57-ras转基因雄性小鼠与BALB/c雌性小鼠杂交,子代通过PCR进行基因型检测,获得C57-ras转基因和野生型小鼠。实验设定2个剂量,75 mg·kg-1MNU和150 mg·kg-1MNU单次腹腔注射给药,共5个组,其中75 mg·kg-1MNU设3个组:(1)转基因小鼠MNU组;(2)野生型小鼠溶媒组;(3)野生型小鼠MNU组;150 mg·kg-1MNU设2个组:(1)转基因小鼠MNU组;(2)野生型小鼠MNU组。其中溶媒组给予同等体积柠檬酸缓冲液,给药后观察29周,观察动物一般症状、生存率、肿物出现时间及大体剖检观察并进行组织病理学检查。结果:75 mg·kg-1MNU给药组小鼠生存率高于150 mg·kg-1MNU。75 mg·kg-1MNU给药后诱发C57-ras转基因小鼠发生淋巴瘤;150 mg·kg-1MNU给药后诱发C57-ras转基因小鼠发生淋巴瘤、肺脏腺瘤和直肠纤维肉瘤。野生型动物溶媒组和野生型动物MNU组均无肿瘤发生。结论:初步验证了自主建立的C57-ras转基因小鼠模型,该模型是临床前药物安全性评价致癌性实验快速替代实验候选模型之一。Abstract Objective： To establish a C57 - ras transgenic mice model for rapid carcinogenicity testing of preclini- cal safety evaluation of drugs in China and carry out a validation study using genotoxic carcinogen N - methyl - N - nitrosourea （MNU）. Methods ： Male C57 - ras transgenic mice were crossed with female BALB/c mice. The off- springs were genotyped using the method of PCR. Both C57 - ras transgenic mice and wild type mice were used for the validation study. There were two dose levels scheduled,75 mg ·kg-1 and 150 mg · kg-1 MNU,which were ad- ministered by single intraperitoneal injection there are five different groups, in which, three groups received 75 mg · kg-1 MNU ：C57 -ras transgenic mice MNU group,wild type mice vehicle control group and wild type mice MNU group; two groups for 150 mg · kg-1 MNU：C57 -ras transgenic mice MNU group and wild type mice MNU group. Vehicle control group was given equal volume of citric acid buffer solution. Both transgenic and wild type mice were observed for the general clinical symptoms, survival rate and time of tumor formation and general observation at nec- ropsy. The collected tissues were undergone histopathological examination to diagnose the tumors. Results： The survival rates of 75 mg · kg-1 MNU groups were higher than those of 150 mg·kg-1 MNU groups. 75 mg · kg-1 MNU could induce the formation of lymphoma in C57 -ras transgenic mice. 150 mg· kg-1 MNU could induce the forma- tion of lymphoma,lung adenoma, and rectum fibrosarcoma in C57 - ras transgenic mice. No tumor was observed in wild type vehicle control group and the two wild type MNU groups. Conclusion： The C57 - ras transgenic micemodel established by ourselves is preliminarily validated by MNU and may be a promising candidate animal model for development of a rapid carcinogenicity testing system for preclinical safety evaluation of drugs.

关 键 词：C57-ras 转基因小鼠 甲基亚硝基脲 验证实验 致癌性实验 

分 类 号：R917[医药卫生—药物分析学]