人羊膜上皮细胞对急性肝衰竭动物模型的治疗效果

Effects of human amniotic epithelial cells therapy on acute liver failure in rats

目的探讨人羊膜上皮细胞（hAECs）对急性肝衰竭（ALF）大鼠模型的治疗效果。方法从剖腹产人胎盘中分离羊膜，经过0．1％Trypsin-0．02％EDTA处理分离hAECs。hAECs用PKH26荧光生物素标记后经腹腔、门静脉、尾静脉三种不同途径注射到大鼠体内，观察细胞在肝脏分布情况，筛选出合适移植途径。选择240—260g雄性大鼠，采用D一氨基半乳糖（D—gal）2g/kg体重腹腔注射法建立ALF大鼠模型。模型制备1d后随机分为对照组和细胞治疗组。治疗组经腹腔直接注射hAECs，对照组注射等量的生理盐水。观察21d内大鼠存活率、肝功能和组织病理变化。结果免疫组化检测显示hAECs表达白蛋白、糖原等肝细胞标记。经门静脉、尾静脉途径移植细胞后可见肝脏内移植细胞定居，经腹腔途径移植的细胞在肝脏内未见分布，但可将多量细胞植入体内且操作简便。由于尾静脉途径移植细胞数量有限，门静脉途径具有创伤且注射后产生微循环栓塞等情况，本课题组选择腹腔直接注射途径实施细胞治疗。细胞移植后大鼠存活率提高，肝功能及细胞因子指标有明显改善，肝脏组织病理逐渐恢复正常。结论通过腹腔途径移植的hAECs对ALF大鼠模型有明显治疗作用。

Objective To investigate the therapeutic effect of human amniotic epithelial cells （hAECs） on acute liver failure （ALF） in rats. Methods hAECs were collected from amnions which were first separated from placenta and then digested with 0.1% Trypsin-0.02% EDTA. PKH26 fluorochrome-labeled hAECs were injected into rats via enterocoelia, portal vein and caudal vein to determine the best injection route. ALF in rats were induced by D-galactosamine （D-gal） via intraperitoneal injection at the dose of 2.0 g/kg body weight. Rats with ALF were randomly divided into two groups ： one group was treated with hAECs through intraperitoneal injection, the other group was treated with normal saline. Then the survival rate of rats, liver function and tissue pathological changes were observed within 21 days. Results hAECs were expressed with albumin, glycogen being markers for hepatocytes, hAECs could be successfully transplanted into rat liver after portal vein and caudal vein injection. However, hAECs injected by the intraperitoneal route were not found in the liver. Since through caudal vein cell count transplanted and found in liver was limited, and portal vein transplantation caused trauma and microcirculation embolism, we chose to inject hAECs through intraperitoneal injection for the treatment of ALF. After hAECs transplantation therapy, the survival rate of rats was increased, the levels of liver function were significantly improved. Liver tissue pathological findings showed that liver injury gradually became within normal range. Conclusion Intraperitoneal injection of hAECs exhibited a significant therapeutic effect on ALF rats.