FLK-1^+胎肝细胞移植治疗急性肝损伤  

Transplantation of fetal liver FLK-1^+ cells in treatment of acute liver injury

作　　者：李娜[1] 丁伟荣[2] 伍柏青[3] 刘婷婷[2] 程洪波[2] 金成豪[2] 

机构地区：[1]南昌大学医学院,江西省南昌市330006 [2]江西省人民医院血液科,江西省南昌市330006 [3]江西省人民医院检验科,江西省南昌市330006

出　　处：《中国组织工程研究》2013年第53期9139-9144,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(30860115);江西省自然科学基金项目(2008GZY0084)资助~~

摘　　要：背景:作者先期研究表明,在鼠胚第17-19天胎肝存在一类FLK-1+细胞,表达胚胎干细胞的特征性标志,并具有多向分化功能。目的:验证胎肝FLK-1+细胞治疗小鼠急性肝损伤的修复作用。方法:免疫磁珠提取及流式细胞仪检测胎肝FLK-1+细胞;RT-PCR检测FLK-1+细胞Oct-3/4、Rex-1基因;肝细胞生长因子和表皮生长因子诱导FLK-1+细胞向肝细胞分化。腹腔注射四氯化碳建立小鼠急性肝损伤模型并随机分为2组:对照组模型小鼠仅输入生理盐水,实验组模型小鼠尾静脉输注诱导分化FLK-1+细胞(1×106细胞),16 h后两组取血测定肝功能,观察64 h小鼠死亡率。结果与结论:胎鼠肝脏FLK-1+细胞高表达Oct-3/4、Rex-1,白蛋白表达的FLK-1+细胞阳性率为0.6%。经肝细胞生长因子诱导3 d后FLK-1不表达,Oct-3/4和Rex-1表达显著下降或消失。经肝细胞生长因子和表皮生长因子诱导后96.38%的FLK-1+细胞表达白蛋白。诱导3d的FLK-1+细胞移植后16 h小鼠血清谷草转氨酶和谷丙转氨酶显著低于对照组(P<0.05),血清白蛋白显著高于对照组(P<0.05);两组血清总胆红素和纤维蛋白原差异无显著性意义(P>0.05);移植组64 h死亡率为61.5%与对照组(80%)差异无显著性意义(P>0.05)。说明胎鼠肝脏肝细胞生长因子和表皮生长因子诱导3 d的FLK-1+细胞移植可较好地改善急性肝损伤的肝细胞功能。BACKGROUND： Previous studies of our research group have shown that FLK-1^＋ cells existed in the mouse fetal liver at embryonic 17-19 days, and they expressed embryonic stem cells and presented multi-differentiation potentials.OBJECTIVE： To evaluate the therapeutic effect of the transplantation of FLK-1^＋ cells derived from mouse fetal liver on acute liver injury in mice.METHODS： The FLK-1^＋ fraction were enriched from the fetal liver with immunomagnetic beads method and detected by flow cytometry. The Oct-3/4 and Rex-1 genes in FLK-1^＋ cells were detected by reverse transcription-polymerase chain reaction. The FLK-1^＋ cells were induced to differentiate into liver cells by hepatocyte growth factor and epidermal growth factor. The model of acute liver injury in mice was established by intraperitoneally injecting carbon tetrachloride 4, and was randomly divided into two groups. In the control group, mouse models with acute liver injury were infused normal saline via tail vein; in the experimental group, mouse models with acute liver injury were infused induced FLK-1^＋ cells （1×106） via tail vein. The blood was collected at 16 hours and liver functions were detected. The mortality of mice was observed at 64 hours. RESULTS AND CONCLUSION： The fetal liver FLK-1^＋ cells highly expressed Oct-3/4 and Rex-1 mRNA, and albumin expressing rate in FLK-1^＋ cells was 0.6%. After induced by hepatocyte growth factor for 3 days, FLK-1 did not express. After induced by hepatocyte growth factor and epidermal growth factor for 3 days, Oct-3/4 and Rex-1 mRNA expression was significantly reduced or disappeared in FLK-1^＋ cells, and 96.38% FLK-1^＋ cells expressed albumin. After FLK-1^＋ cells induced for 3 days were transplanted to mouse models with acute liver injury for 16 hours, the serum glutamic-oxalacetic transaminease and glutamic-pyruvic transaminase were significantly lower （P 〈 0.05）, but serum albumin was significantly higher than the control group （P 〈 0.05）. However,

关 键 词：器官移植 肝功能损伤 FLK-1^+细胞 细胞移植 肝细胞生长因子 表皮生长因子 谷草转氨酶 谷丙转氨酶 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]