永生化人肝细胞治疗小鼠急性肝衰竭的实验研究  

Treatment of acute liver failure in mice by immortalized human hepatocytes

作　　者：房青[1,2] 高福云[2] 徐梅[2] 张文健[2] 叶丽亚[2] 许世清[2] 许亚平[2] 李成辉[3] 娄晋宁[2] 

机构地区：[1]北京协和医学院研究生院,100021 [2]北京中日友好医院临床医学研究所,100029 [3]北京中日友好医院手术麻醉科,100029

出　　处：《中国医药生物技术》2010年第4期252-256,共5页Chinese Medicinal Biotechnology

基　　金：国家高新技术发展研究计划(863计划)(2006AA02Z482)

摘　　要：目的评价微载体培养的永生化人肝细胞(immortalized human hepatocytes,IHH)对裸鼠急性肝衰竭(acute liver failure,ALF)的治疗作用。方法采用腹腔注射四氯化碳(CCl4)建立裸鼠ALF模型,将微载体培养的IHH注射于裸鼠腹腔内进行腹膜透析治疗,并设未治疗组和空微载体治疗组,比较各组动物的肝功能损伤指标,肝组织病理和动物生存率。结果腹腔注射CCl4后24h,所有动物的转氨酶升高达正常值10倍以上,未治疗组和空微载体治疗组动物48h内全部死亡,肝组织检查发现肝细胞广泛坏死;而采用IHH微载体治疗组,48h的动物生存率为83.3%,持续14d后转为长期存活。存活的动物转氨酶降至正常水平,肝组织检查显示肝细胞坏死显著减少,免疫组织化学染色显示腹腔微载体上肝细胞仍存活,并表达人肝细胞特异蛋白α1-抗胰蛋白酶(α1-antitrypsin,α1-AT)。结论采用微载体培养的IHH腹腔透析治疗小鼠ALF是有效的,表明IHH适合作为生物人工肝的细胞材料。Objective To evaluate the efficiency of microcarriers cultured with immortalized human hepatocytes （IHH） on treatment of acute liver failure （ALF） in animal model. Methods ALF model was established by intraperitoneal injection of CCl4 in nude mice. The microcarriers cultured with IHH were intraperitoneally injected to ALF animal for dialysis,the ALF animals treated with empty microcarriers were used as control. The efficiency of the dialysis was evaluated by plasma aminotransferase,histopathological examinations and survival rate of ALF mice. The immunohistochemical staining for α1-antitrypsin in microcarriers with IHH was performed to analyze their viability and function. Results The plasma aminotransferase was increased over 10 times in ALF mice or in ALF mice treated with empty microcarriers as compared with that in normal mice 24 hours after CCl4 administration,all mice with ALF without any treatment or treated with empty microcarriers died within 48 hours and extensive hepatocellular necrosis was demonstrated by histopathological examinations. Although the plasma aminotransferase was similar to those control groups 24 hours after CCl4 administration,the survival rate of ALF mice treated with IHH-microcarriers was 83.3% within 48 hours after CCl4 administration and maintained to long-term survival. The histopathological examinations showed that hepatocyte necrosis was significantly reduced in ALF mice treated with IHH-microcarriers as compared with controls and immunohistochemical staining indicated that IHH on microcarriers was viable and expressed α1-antitrypsin. Conclusion The intraperitoneal dialysis with microcarriers cultured with IHH is effective for ALF therapy in vivo,suggesting the IHH is suitable for cell material of bioartificial liver.

关 键 词：肝功能衰竭 急性 肝 人工 腹膜透析 

分 类 号：R575.3[医药卫生—消化系统]