胚肝细胞移植治疗Wilson病的初步研究

Amelioration of disturbed copper metabolism in TX mouse:a Wilson disease model, by fetal hepatocytes transplantation

目的 探讨胚肝细胞脾内移植对TX小鼠铜代谢紊乱的改善情况。方法 组织块酶消化法分离培养DL小鼠胚肝细胞 ,以Hoechst3332 4进行荧光标记后作为供体对TX小鼠进行脾内移植 ,检测移植后TX小鼠的血清铜蓝蛋白和血清铜水平 ,并进行病理学检查。结果 移植后 2周内 ,TX小鼠的铜蓝蛋白由 9 0U/L逐渐上升至 14 5U/L、血清铜由 0 3mg/L上升至 0 6mg/L(P <0 0 5 ) ,受体肝组织内可见供体细胞定居并呈结节样增生。结论 胚肝细胞脾内移植后可在受体肝组织中定居增生 ,并有效改善TX小鼠的铜代谢障碍 ,为Wilson病的治疗提供了新的思路。

Objective To investigate the possibility of amelionation of the disturbed metabolism by fetal hepatocytes transplantation in toxic milk mouse, a model of Wilson disease (WD). Methods The donor hepatocytes were harvested from the fetal livers of a normal pregnant DL mouse (D=13 days). The cells were labeled with fluorochrome Hoechst 33 342 and then transplanted into the spleens of TX mice. The serum copper and ceruloplasmin concentrations of the mice were determined after transplantation and histological investigation performed. Results Within 2 weeks after transplantation, the serum ceruloplasmin and copper concentrations increased gradually, (9.0±1.6) U/L vs. (14.5±3.8) U/L; (0.3±0.1) mg/L vs. (0.6±0.2) mg/L respectively, P <0.05, which ultimately rose from about 30% of the normal level to nearly 60%. Pathological examination showed that the donor cells could be visualized in mass of 5 to 8 cells in the liver of the recipients when sacrificed after transplantation. Conclusions The hereditary abnormalities of copper metabolism in TX mice could be corrected partially by intrasplentic transplantation of normal fetal hepatocytes. This approach suggested a new way for perspective clinical therapy of WD.