骨髓间充质干细胞移植对大鼠暴发性肝功能衰竭治疗作用的初步观察

The preliminarily investigation of therapeutic effects of mesenchymal stem cells on rat fuiminant hepaticfailure

目的探讨骨髓间充质干细胞（MSC）移植对大鼠暴发性肝功能衰竭（FHF）的治疗作用。方法全骨髓细胞贴壁培养筛选法分离纯化大鼠MSC；四氯化碳灌胃制作大鼠FHF模型，分为实验组和模型对照组，各20只，8只灌服等剂量0．9％氯化钠溶液作为空白对照组。将4’，6-二脒基-2-苯基吲哚（DAPI）标记的大鼠MSc1．0×10^6从尾静脉移植入实验组和空白对照组，等剂量0．9％氯化钠溶液尾静脉注入模型对照组；分别于术后7、14d处死各组部分大鼠，评价实验组与模型对照组大鼠一般状况、生存率、肝功能、TNF-a水平、肝脏病理、MsC归巢至肝脏的情况。正态分布资料采用两个独立样本t检验，非正态分布资料采用非参数检验。结果术后3d开始，实验组存活大鼠一般情况比模型对照组好转更加明显；术后7d，实验组与模型对照组各有15只和8只大鼠存活，差异有统计学意义（X^2-4．122，P〈0．05）；术后7d以及14d，实验组与模型对照组相比，肝功能指标、TNF-a水平均差异均有统计学意义（均P（0．05），病理学检查肝组织炎性反应改善情况实验组均更为明显；术后实验组MSC归巢细胞逐渐增多，空白对照组只观察到很少量归巢细胞。结论MSC经尾静脉同种异体移植，可以归巢到肝功能衰竭大鼠肝脏，改善大鼠免疫和肝脏组织炎性反应坏死状态，促进FHF大鼠肝功能的恢复，对FHF具有明显的治疗作用。

Objective To investigate the therapeutic effects of mesenchymal stem cells （MSC） on rat fulminant hepatic failure （FHF）. Methods The rat MSC were separated and purified by adherent culture of whole bone marrow cells. The rat FHF models were established by CC14 intragastric administration. The rats were divided into experimental group （n= 20） and model control group （n=20）. And the same dose of saline was administered to rats as normal controls （n= 8）. Dosage of 1.0 × 10^6 4＇, 6-diamidino-2-phenylindote （DAPI） labeled MSC were transplanted into rats inexperimental group and normal control group through caudal veins, and the same dose of saline was given intravenously in model control group. Part of rats in each group were sacrificed after 7 days and 14 days of injection to evaluate the general condition, survival rate, liver function, tumor necrosis factor （TNF）-a level, liver pathology and MSC homing to the liver between experimental group andmodel control group. Normal distribution data were compared by independent-sample t test and non- normal distribution data were analyzed by non-parameter test. Results After 3 days of injection, the general condition of experimental group were better than the model control group. After 7 days of injection, therewere 15 and 8 survival rats, the survival rates were statistically different between experimental group and model control group （X^2 =4. 122, P〈0.05）. After 7 days and 14 days ofinjection, the liver function and TNF-a levels were statistically different between experimental group and model control group （both P d0.05）, and liver pathology improvement in experimental group was more significant than model control group. DAPI labeled cells increased after transplantation in experimental group, while few DAPI labeled cells were observed in normal control group. Conclusions MSC can home to liver of FHF rats after MSC allogeneic transplantation through caudal veins, which can improve liver immunity and liver tissue necroinflammation, and facilitate recovery of liver function. Therefore, it is demonstrated that MSC transplantation has obvious therapeutic effect on rat FHF.