猕猴自体骨髓干细胞移植治疗急性肝损伤的实验研究

Application of Autologous Bone Marrow Stem Cells for the Treatment of Acute Liver Necrosis in Monkey

目的探讨自体骨髓干细胞肝门静脉移植对猕猴急性肝损伤的治疗作用和为人类肝损伤治疗提供依据和方法。方法猕猴10只,随机分为实验组和对照组,每日皮下注射40%CCL40.8ml/kg,连续注射4d,经肝功能生化指标检测及肝穿组织病理学观察确认肝损伤后,分离实验组猴的自体骨髓单个核细胞,按3×108细胞/kg将骨髓细胞植入肝门静脉,植入后60d进行肝功能相关指标检测及组织病理学观察。结果连续注射CCL44d后,血清谷草转氨酶(AST)、谷丙转氨酶(ALT)、球蛋白(GLOB)(P<0.001),而总蛋白(TP)和白蛋白含量无变化(P>0.05),肝组织结构紊乱,大量肝细胞坏死,部分细胞变性,水肿,汇管区及周围有大量炎性细胞浸润。骨髓细胞肝门静脉移植后60d,血清AST、ALT活性和GLOB、BIL含量显著低于对照组(P<0.05),而TP、ALB、LDL含量高于对照组(P<0.05),病理组织学观察结构清楚,而对照组有纤维增生和肝细胞有脂肪变性。结论自体骨髓细胞肝门静脉移植能对猕猴急性肝损伤有明显治疗作用,可改善急性肝损伤猕猴的肝功能及组织结构,提示该方法可能对人类急性肝损伤有一定治疗意义。

Objective A monkey model was used to determine the feasibility of treating acute liver necrosis by injecting autologous bone marrow stem cells via the portal vein. Methods Ten 7-10 years old monkeys were induced to develop liver necrosis by daily subcutaneous injection with 40% CCL4 0.8 ml/kg for 4 days. The animals were then randomely divided into experimental and control groups. In the experimental group, autologous marrow stem cells（3×10^8） were tranplanted into liver through the injection of cells via the portal vein. Enzymes related to the liver function, proteins, lipids and bilirubin content in plasma were analyzed by an automatic biochemical analyzer. The change of liver functions was monitored by both histopathologic and biochemical methods 60 days after transplantation. Results After CCL4 injection, hepatocyte necrosis was noted. An increased in GLOB, TBIL, ALT and AST activity （P〉0.001） was also observed. The levels of TP, ALB, and HDL were not changed （P〉0.05） in plasma. Sixty days after bone marrow transplantation, the activity of AST and ALT, and the levels of GLBO and BIL were lower than the control group（P〈0.05）. However, the levels of TP, ALB, and LDL were higher than the control group （P〈 0.05）. Conclusion A monkey liver necrosis model was successfully established by injecting the monkey with 40% CCL4 injection. Autologous bone marrow stem cells transplanted through the portal vein is feasible, and the transplanted stem cells can repopulate in the necrotic liver.