肝干细胞移植治疗大鼠暴发性肝功能衰竭的实验研究  

Study on hepatic stem cell transplantation in fulminant hepatic failure in rat

作　　者：杜智[1] 李涛[1] 张金卷[1] 朱争艳[1] 高英堂[1] 王毅军[1] 聂福华[1] 

机构地区：[1]天津市第三中心医院肝胆外科,卫生部人工细胞工程技术研究中心,天津300170

出　　处：《生物医学工程与临床》2006年第6期346-349,共4页Biomedical Engineering and Clinical Medicine

基　　金：国家863课题(2001AA216051)

摘　　要：目的探索肝干细胞移植对暴发性肝功能衰竭大鼠的治疗效果。方法以二乙酰氨基芴(15mg/kgBW)联合2/3肝切除建立雄性大鼠肝干细胞增殖模型,以改进Seglen胶原酶原位灌注及Percoll密度梯度离心方法分离、纯化肝干细胞,经肝脏注射移植(4×106细胞)治疗由D-氨基半乳糖诱导的暴发性肝功能衰竭雌性大鼠,移植前及移植后24、48、72h及1周取血测定肝功能指标如血氨、ALT、TBiL,移植后1、2、4周取受体肝脏组织应用PCR方法进行性别决定因子检测。结果肝干细胞移植可延长暴发性肝功能衰竭大鼠的中位生存时间(P<0.05),移植后肝干细胞移植组大鼠血清ALT、TBiL及血氨水平明显下降,其测定值各时间点组间比较,治疗组明显低于对照组(P<0.01);肝脏病理损伤减轻,肝干细胞移植2周后雌性受体肝脏组织中性别决定因子呈阳性表达,且随时间延长而表达增强。结论肝干细胞移植可延长暴发性肝功能衰竭大鼠中位生存时间,改善肝功能及病理指标。Objective To study the therapeutic effect of hepatic stem cell （HSC） transplantation in fulminant hepatic failure （FHF） of rat. Methods The HSC proliferation model were induced in male rats with administration of 2-AAF 15mg/kg BW. combined with 2/3 PH on day 5 ,and then administered 2-AAF daily up to 11 days after operation. Then the HSC were isolated and purified by collagenase（type Ⅰ ） peffusion method and Percoll gradient centrifugation（2 000 r/min,20 min）. While 24 hours after incubation ,0.4 ml purified cell suspension （107/ml） was injected directly into the liver in female rat with FHF induced by D- galactosamine （D-GlaN, 1 200 mg/kg BW）. The serum ALT,TBiL and blood ammonia levels were measured pre- and posttransplantation at different time points. To identify the male HSCs in female rats, sex determining region of the Y（Sry） chromosome was examined by PCR technique at 1,2,4 weeks after transplantation. Results The survial time of HSC in transplantation group was longer than that of control group （122.10 hours vs 68.50 hours,P 〈 0.05）. Biochemical parameters were decline after HSC transplantation as the time goes by, and lower than that of the control group at each time points（P 〈 0.01）. The hepatocyte necrosis in rat liver sections was improved after HSC transplantation, small oval shape nuclear cells were proliferated increasingly in portal area or central vein region and Sry was detected in female recipient rat liver 2 weeks after transplantation. Conclusion Stem cell transplantation could prolong the suvival time of recipients ,correct their biochemical disorders and liver pathological injury of rats with FHF.

关 键 词：肝干细胞 细胞移植 暴发性肝功能衰竭 

分 类 号：R-332[医药卫生]