利用bcl-2抗Fas凋亡特性联合Fas抗体促进人鼠嵌合肝中人肝细胞的增殖  

Repopulation of bcl-2 gene transfection on HepaRG cells in an animal model of severe combined immunodeficient mice with chimeric human liver using mouse-specific anti-fas antibody

作　　者：蒋黎[1] 兰林[1] 李俊刚[1] 王宇明[1] 刘国栋[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《重庆医学》2009年第19期2445-2447,2450,I0003,共5页Chongqing medicine

基　　金：国家自然科学基金资助项目(30470882);国家863计划项目(2006AA02A140)

摘　　要：目的移植bcl-2转基因HepaRG细胞到小鼠体内,并联合使用Jo2抗体诱导鼠肝细胞凋亡策略,促进人鼠嵌合肝中人肝细胞的增殖。方法SCID鼠经脾移植2×106bcl-2转基因HepaRG细胞,移植后1d始腹腔注射0.2mg/kgJo2抗体,每周1次,持续10周为实验组;移植未转染bcl-2基因的HepaRG细胞,同样给予Jo2抗体腹腔注射为对照1组,移植未转染bcl-2基因的HepaRG细胞,未腹腔注射Jo2抗体为对照2组,建立人鼠嵌合肝动物模型。从2周开始,后4、8、12、16、20、24周,分别采用免疫组化、免疫荧光和逆转录聚合酶链反应(RT-PCR)检测不同时相点实验组和对照组嵌合肝中肝组织人清蛋白以及人清蛋白mRNA,并用酶联免疫法(ELISA)定量检测鼠血清人清蛋白的含量。结果实验组和对照组小鼠均能存活至24周,鼠肝组织和血清中均能检测到清蛋白的表达。实验组肝组织和血清人清蛋白的表达均可持续到24周,人血清清蛋白高峰值为95.32ng/mL,出现在16周;而对照1组肝组织和血清人清蛋白的表达可持续到20周,人血清清蛋白高峰值为42.37ng/mL,出现在12周;对照2组肝组织和血清人清蛋白的表达可持续到12周,人血清清蛋白高峰值为22.91ng/mL,出现在8周。结论利用bcl-2抗Fas凋亡特性,转染bcl-2基因的HepaRG细胞移植并联合小剂量Jo2抗体腹腔注射SCID鼠,有利于人鼠嵌合肝中人肝细胞的增殖和存活。Objective Repopulation of severe combined immunodeficient （SCID）mouse liver with bcl-2 gene transfeetion on HepaRG cells was studied following induction of mouse hepatocyte apoptosis using an anti-mouse agonistic fas monoelonal antibody （Jo2 mAb）. Methods SCID mice were transplanted with 2 × 10^6 bcl-2 gene transfection on HepaRG cells intrasplenically and treated once a week for 10 weeks with 0.2mg/kg Jo2 mAb intraperitoneally consistently for experimental group. Mice were transplanted with non-gene transfection on HepaRG cells and treated with Jo2 mAb for control group one,and mice were transplanted with non-gene transfection on HepaRG cells and non-treated with Jo2 mAb for control group two. From 2 to 24 weeks after transplantation,human hepatocytes repopulation were characterized by S-P immunohistochemistry and immunofluorescence staining for human albumin,and reverse transcription-polymerase chain reaction （RT-PCR） to detect human albumin mRNA in mice liver. Meanwhile, human serum albumin was detected by enzyme linked immunosorbent assay（ELISA） in mice serum at the same time. Results Mice of experimental group and control group were survival to 24 weeks,liver and serum of which were detected human albumin. In experimental group, human albumin was detectable in mice liver and serum up to 24 weeks after transplantation, but 20 weeks for control group one and 12 weeks for control group two only. High-peak of human albumin appearanced at 16th week for experimental group, 12 weeks for control group one and 8th week for control group two. The quantity of human serum albumin at most of diffirent group were 95.32ng/ml, 42. 37ng/ml, and 22. 91ng/ml individually. Conclusion With bcl-2 gene of anti Fas peculiarity, transplantation of bcl 2 gene transfection on HepaRG cells into SCID mice using mouse-specific anti-fas antibody,can be promote survival and proliferation of human hepatocytes in chimeric mice.

关 键 词：BCL-2 HepaRG细胞 Fas抗体/Jo2 MAB 移植 人鼠嵌合肝/动物模型 

分 类 号：R392.4[医药卫生—免疫学] R365.575[医药卫生—基础医学]