三种不同剂量DEN诱导大鼠原发性肝癌模型的比较  

Comparison of three different doses of DEN induced primary liver cancer models in rats

作　　者：张日云 吴凤兰 毛德文[2,3] 王明刚[2,3] 裴浩 李飞燕 ZHANG Riyun;WU Fenglan;MAO Dewen;WANG Minggang;PEI Hao;LI Feiyan(Guangxi University of Chinese Medicine,Nanning 530200,China;the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China;Guangxi Key Laboratory of Molecular Biology of Prevention and Treatment of Traditional Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学,南宁530200 [2]广西中医药大学第一附属医院,南宁530023 [3]广西中医药防治医学分子生物重点实验室,南宁530023

出　　处：《中国实验动物学报》2025年第2期169-179,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(82274434);广西中青年教师科研基础能力提升项目(2023KY0313);广西中医药大学第一附属医院博士基金启动课题(2020BS003)。

摘　　要：目的 使用3种不同剂量的二乙基亚硝胺(diethylnitrosamine, DEN)建立大鼠原发性肝癌(primary liver cancer, PLC)模型,旨在构建一个高效、稳定、经济的PLC动物模型。方法 将雄性SD大鼠45只,随机分为4组,分别为正常组、低剂量组(50 mg/kg)、中剂量组(70 mg/kg)、高剂量组(200 mg/kg)。正常组6只,其余组各13只。正常组不做任何处理。低剂量组第1~4周,每周腹腔注射2次,第5~12周,每周注射1次;中剂量组每周腹腔注射1次,连续16周;高剂量组仅第1周给药1次。随后观察各组大鼠至16周。通过生存率、病理学检测、生化检测、肝脾指数计算、免疫组化、酶联免疫吸附测定法(ELISA)等检测方法验证模型的建立及优化评价。结果 正常组大鼠存活率为100%,低剂量组为46.15%,中剂量组为69.23%,高剂量组为84.61%。肉眼观察正常组大鼠肝组织未见异常;低剂量组大鼠肝颜色变深,表面较粗糙,可见少量癌结节,质地稍硬;中剂量组大鼠肝表面粗糙,可见多个小的癌结节及散在块状占位性结节,质地坚硬;高剂量组大鼠肝颜色变浅,表面稍粗糙,未见明显癌结节。苏木素-伊红(HE)染色显示低、中剂量组肝组织结构紊乱,细胞异型性大,肿瘤细胞形成,高剂量组肝小叶结构不清晰,肝细胞呈不同程度的水肿、变性、坏死,未见明显肿瘤细胞形成。与正常组对比,低、中、高剂量组血清谷丙转氨酶(alanine aminotransferase, ALT)、谷草转氨酶(aspartate aminotransferase, AST)、总胆红素(total bilirubin, TBIL)均有升高;低剂量组ALT、AST升高显著(P<0.05),差异具有显著性,中剂量组ALT、AST、TBIL均显著升高(P<0.05),差异具有显著性,高剂量组的ALT、AST、TBIL虽有升高,但升高不明显,差异无显著性(P>0.05);与正常组相比,低剂量组凝血功能国际标准化比值(international normalized, INR)明显增高(P<0.05),差异具有显著性,活化部分凝血活酶时间(activated partial thromboplastObjective Three different doses of diethylnitrosamine(DEN)were used to establish a rat primary liver cancer(PLC)model to establish an efficient,stable,and economical animal model of PLC.Methods Forty⁃five male SD rats were randomly divided into four groups:normal group,DEN 50 mg/kg dose group(low dose group),70 mg/kg dose group(medium dose group),and 200 mg/kg dose group(high dose group).There were 6 animals in the normal group and 13 animals in each of the other groups.The normal control group received no treatment.The model group and low dose groups were injected intraperitoneally twice a week during weeks 1~4 and once a week during weeks 5~12;the medium dose group was injected intraperitoneally once a week for 16 consecutive weeks;and the high dose group was administered only once in the first week.The rats in each group were then followed for 16 weeks.The establishment of the model and optimal evaluation were verified by survival rate,pathological tests,biochemical tests,liver and spleen index calculation,immunohistochemistry,enzyme⁃linked immunosorbent assay(ELISA),and other assays.Results The survival rate was 100%in the normal group,4615%in the low dose group,6923%in the medium dose group,and 8461%in the high dose group.The liver tissues of the rats in the normal group showed no abnormality to the naked eye;the liver of the rats in the low dose group became darker in color,rougher in surface,with a small number of cancerous nodules and slightly hard texture;the liver of the rats in the medium dose group was rough in surface,with several small cancerous nodules and scattered massive occupying nodules and hard texture;The liver of rats in the high dose group became lighter in color,slightly rougher in surface,with no obvious cancerous nodules;HE staining showed that the liver tissues of rats in the low and medium dose groups were structurally disorganized,with large cellular heterogeneity and tumor cells.HE staining showed that the liver tissues of rats in the low and medium dose groups were structurally

关 键 词：原发性肝癌 二乙基亚硝胺 造模方法 大鼠 模型优化 

分 类 号：Q95-33[生物学—动物学]