二乙基亚硝胺诱发肝癌前病变大鼠模型的建立  

Def inition of diethylnitrosamine-induced rat hepatic precancerous lesions with atypia index

作　　者：宋艺君[1] 刘晓婷[1] 冯丽莉[1] 王湘[1] 白荟芳[1] 田民强[1] 张微[1] 祖承哲[2] 赵欣[2] 蔡大勇[2] 王玥琦[1] 

机构地区：[1]北京中医药大学基础医学院,北京市100029 [2]中国医学科学院药用植物研究所,北京市100193

出　　处：《世界华人消化杂志》2012年第27期2562-2575,共14页World Chinese Journal of Digestology

基　　金：重大新药创制科技重大专项基金资助项目;No.2009ZX09502-017;教育部重点研究基金资助项目;No.108019;北京中医药大学创新团队基金资助项目;No.2011-CXD-04~~

摘　　要：目的:研究二乙基亚硝胺诱发大鼠肝癌前病变的时效关系与量效关系.方法:Wistar♂大鼠100只,体质量均衡随机分5组;每84h1次分别腹腔注射二乙基亚硝胺0、25、50、75、100mg/kg造模.每组体质量排序分10小组,年龄配对;造模时程最短0d,间隔14d延长,最长126d,灌注固定肝脏.常规石蜡包埋,6μm切片,HE染色,观察病变性质后显微照相;使用Image-Pro Plus软件,分别形态计量10、20、40倍物镜下汇管区(R1)、肝索(R2)、肝细胞核(R3)的体积构成比,观察每10个高倍镜视野下的核分裂像;计算各动物异型性指数[Ei=(1-R1)-1·(R3)·(R2)-1].Prism 4软件回归"二乙基亚硝胺导致异型性指数"的时效曲线及曲线下面积,进而导致曲线下面积的量效关系.结果:以异型性指数(肝细胞核/浆体积比校正值),回归0、25、50、75、100mg/kg二乙基亚硝胺造模的半效时程(95%CI)分别为70347(0-)、1734(937-3211)、1536(948-2490)、1530(890-2632)、1183(955-1466)h,曲线下面积分别为0.0064、0.0084、0.0123、0.0165、0.0167[异型性指数·log(h)].以曲线下面积,回归二乙基亚硝胺诱发肝细胞癌前病变的半效剂量为48.255mg/kg.50.000mg/kg造模各时程的异型性指数与核分裂像总数之间呈直线正相关(y=0.0023x-0.0056,r=0.9217,n=10,P<0.01).结论:以异型性指数为指标,每84h腹腔注射1次二乙基亚硝胺,诱发♂大鼠肝细胞癌前病变的最佳造模剂量为48.255mg/kg(约50.000mg/kg),最佳造模累积连续时程为64d.核分裂像计数验证了异型性指数的临床预警价值.AIM：To def ine diethylnitrosamine-induced hepatic precancerous lesions with atypia index in rats.METHODS：After normalization of the body weight,100 male Wistar rats were equally and randomly divided into f ive groups and were intraperitoneally injected with different doses of diethylnitrosamine in normal saline（0,25,50,75,100 mg/kg） twice weekly.The rats in each group were further divided into 10 subgroups with endpoints ranging from d00 to d126 with an interval of 14 days.Liver samples were fixed,sectioned,and stained with hematoxylin and eosin.After being evaluated pathologically,the volume ratios of portal triad（R1）,hepatic cord（R2） and hepatic nucleus（R3） were obtained under an Olympus microscope with 10-,20-,and 40-amplification,respectively,by use of Image-Pro Plus software.Hepatic mitotic f igures were counted under high magnification.The atypia index of hepatocytes was calculated as the volume ratio of nucleus to cytoplasm in precancerous lesions using the formula [（1-R1）-1·（R2）-1·（R3）].By using Graph Pad Prism version 4 for Windows,the half effective durations and the area under curve of the atypia indexes were regressed in Sigmoidal time-response（variable slope） from the logarithm of endpoint hours [11.4 ＋ Time（d） ×24 h/d] for each of 5 doses,so did the half effective doses of the area under curves from the logarithm of dosages [10 ＋ Dose（mg/kg） ×1 000（μg/kg）].The atypia indexes were regressed linearly with the mitotic counts to conf irm their specif icity in predicting hepatic carcinogenesis.RESULTS：From the atypia index of hepatocytes（corrected volume ratio of nucleus to cytoplasm）,the half effective durations of diethylnitrosamine at 0,25,50,75,and 100 mg/kg to induce hepatic precancerous lesions were 70 347,1 734,1 536,1 530 and 1183 h,respectively.The areas under curves were 0.0064,0.0084,0.0123,0.0165 and 0.0167 [（atypia index） × log（h）],respectively.From the area under curve,the half effective dose of diethylnitrosamine to in

关 键 词：二乙基亚硝胺 肝癌前病变 异型性指数 核分裂像 形态计量 

分 类 号：R735.7[医药卫生—肿瘤]