L02细胞在倒千里光碱处理大鼠肝中的增殖  

Repopulation of human L02 hepatocytes in tolerant rats treated with retrorsine

作　　者：林沪[1] 毛青[1] 王宇明[1] 蒋业贵[1] 蒋黎[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》2007年第4期346-352,共7页World Chinese Journal of Digestology

基　　金：国家自然科基金.No.30271173

摘　　要：目的:应用倒千里光碱(retrorsine,Rts),研究正常人L02肝细胞移植到具有正常免疫活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人L02肝细胞,诱导胎鼠对人L02肝细胞产生免疫耐受,出生3 wk后分别ip Rts或生理盐水,再经脾移植DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindo- carbocyanine perchlorate)染色后的人L02肝细胞,建立人鼠嵌合肝动物模型．采用免疫荧光、逆转录聚合酶链反应(reverse transcription-pOIymerase chain reaction,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、特异性人增殖细胞核抗原(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:实验组于移植后1 wk-6 mo,在荧光显微镜下观察到人L02肝细胞在鼠肝内的动态分布,而对照组为移植后1-10 wk;实验组移植后2 wk-6 mo,大鼠检测出人白蛋白、人白蛋白mRNA及特异性人增殖细胞核抗原PCNA．而对照组分别于移植后2-8 wk检测出白蛋白及人白蛋白mRNA,移植后2 wk-6 mo检测出特异性人PCNA．以上各组阳性细胞的出现均以移植4 wk最多．PCNA阳性细胞的数密度和面积密度均显示,特异性人PCNA组与其对照组比较有统计学差异(P＜0．05)．结论:Rts对移植L02细胞有明显的增殖作用, L02细胞在人鼠嵌合肝动物模型中存活并增殖6mo以上．d AIM： To investigate whether human L02 hepatocytes can repopulate after transplantation into the normal and immunocompetent rats treated with retrorsine.METHODS： Human L02 hepatocytes were injected into the peritoneal cavities of fetal Sprague Dawley rats to induce immune tolerance. The 3-week-old rats were injected with retrorsine or normal saline. After DiI （1, 1＇-dioctadecyl-3, 3, 3＇, 3＇-tetramethylindocarbocyanine perchlorate） staining, L02 hepatocytes were transplanted into the 7-week-old rats via the spleen. Immunofluorescent staining, reverse transcriptionpolymerase chain reaction （RT-PCR）, S-P immu- nohistochemistry, and DiI staining were used to detect human albumin, human albumin mRNA and specific proliferating cell nuclear antigen （PCNA） in rat livers. The distribution of human L02 hepatocytes was observed under fluorescent microscope. The number density and area density of PCNA were obtained with the help of computer-assisted image analyzer.RESULTS： Dynamic distribution of human L02 hepatocytes in rat livers was observed from the 1^st week to 6^th month after transplantation, but it was from the 1^st to 10^th week in the controls. Human albumin and albumin mRNA （241 bp） were detected from the 2^nd week to the 6^th month after transplantation, but they could only be detected from the 1^st to 8^th week in the controls. The highest expression of human albumin and albumin mRNA appeared at the 4^th week. Specific human PCNA was detected in rat livers from the 2^nd week to the 6^th month after transplantation, but it could only be examined from the 2^nd to 6^th week in the controls. The number of PCNA-positive cells reached the peak at the 4^th week. The number density and area density of PCNA were significantly different between specific PCNA group and its control group （P 〈 0.05）.CONCLUSION： Retrorsine is able to promote the repopulation of transplanted human L02 hepatocytes, and L02 hepatocytes can survive more than 6 months, and repopulate at the same time.

关 键 词：肝细胞 嵌合肝 移植 大鼠 增殖 倒千里 光碱 

分 类 号：R575[医药卫生—消化系统]