L02细胞在2-乙酰氨基芴处理大鼠肝中的增殖  

Repopulation of human L02 hepatocytes in tolerant rats treated with 2-acetaminofluorene

作　　者：林沪[1] 毛青[1] 王宇明[1] 蒋业贵[1] 邓国宏[1] 蒋黎[1] 李俊刚[1] 刘国栋[1] 

机构地区：[1]中国人民解放军第三军医大学西南医院感染病研究所,重庆市400038

出　　处：《世界华人消化杂志》2006年第23期2281-2287,共7页World Chinese Journal of Digestology

基　　金：国家自然科基金资助项目;No.30271173~~

摘　　要：目的:应用2-乙酰氨基芴(2-acetaminofluorene, 2-AAF),研究L02细胞移植到具有正常免疫性的大鼠肝内的存活与增殖情况．方法:SD大鼠出生前宫内ip正常人L02肝细胞,诱导胎鼠对L02肝细胞产生免疫耐受,出生2 wk时分别ip小(1．2 mg／kg)、中(13 mg／kg)、大剂量(30 mg／kg)2-乙酰氨基芴或生理盐水,隔日行2／3肝切除术,同时经脾移植DiI染色后的人L02肝细胞,建立2-AAF／PH肝细胞增殖模型．采用免疫荧光、RT-PCR、免疫组化、DiI荧光示踪等方法,在不同时相分别检测人白蛋白、人白蛋白mRNA、增殖细胞核抗原(PCNA)以及在荧光显微镜下观察人L02肝细胞在鼠肝内的分布,并采用计算机图像分析系统分析不同时相PCNA阳性细胞的数密度和面积密度．结果:2-乙酰氨基芴实验组(小、中、大剂量)的各项检测,均与对照组无明显区别;于移植后1,2,4,6,8,10 wk在荧光显微镜下观察到人L02肝细胞在鼠肝内的动态分布;移植后2, 4,6,8 wk大鼠均检测出人白蛋白及人白蛋白mRNA;移植后2,4,6 wk检测出特异性人增殖细胞核抗原PCNA,移植后1,2,4,6,8 wk检测出非特异性PCNA．结论:2-AAF／PH模型中,2-乙酰氨基芴对移植L02细胞无明显增殖作用;L02细胞在2-乙酰氨基芴处理人鼠嵌合肝动物模型中存活10 wk,产生白蛋白功能持续8 wk．AIM： To investigate whether human L02 hepatocytes can repopulate after implantation to the normal, immunocompetent rats treated with 2-acetaminofluorene. METHODS： Human L02 hepatocytes were injected into the peritoneal cavities of fetal Sprague Dawley rats to induce immune tolerance. The 2-week rats were injected with different doses of 2-acetaminofluorene or normal saline, and received partial hepatoectomy. After 1, 1＇-Diocta- decyl-3, 3, 3＇, 3＇-tetramethylindocarbocyanine perchlorate （DiI） staining, the hepatocytes were implanted to the 3-week rats via the spleen. Immuno-fluorescent staining, reverse transcription-polymerase chain reaction （RT-PCR）, S-P immunohistochemistry, and DiI staining were used to detect human albumin, human albumin mRNA and specific proliferating cell nuclear antigen （PCNA） in the rat liver. The distribution of human L02 hepatocytes was observed under fluorescent microscope. The number and area density of PCNA-positive cells were quantified with the help of computer-assisted imgine analyzer. RESULTS： All the examined indicators were not significantly different between the rats treated with 2-acetaminofluorene and normal saline. Dynamic distribution of human L02 hepatocytes in the rat livers was observed from wk 1 to 10 after implantation. Human albumin and human albumin mRNA were detected from wk 2 to 8 after implantation and the peak content appeared at the 4th wk. Specific human PCNA was detected in the rat livers from wk 2 to 6 after implantation. There were the most PCNA-positive cells at the 4th wk. Nonspecific PCNA was detected in the rat livers from wk I to 8 after implantation. ONCLUSION： Human L02 hepatocytes can not significantly repopulate after implantation to the normal, immunocompetent rats treated with 2-acetaminofluorene. Human L02 hepatocytes can survive for 10 wk after implantation, and exhibit unique hepatic biochemical functions for 8 wk.

关 键 词：嵌合肝 L02肝细胞 动物模型 移植 大鼠 

分 类 号：R657.3[医药卫生—外科学]