大鼠移植骨髓细胞向肝细胞转化的实验研究

目的 探讨体内骨髓细胞向肝细胞转化的可行性。方法 将雌性SD大鼠随机分为3组，每组15只。①R BMT(全身照射 骨髓移植)；②2—AAF R BMT；③2—AAF PH(部分肝切) BMT。进行交叉性别骨髓细胞移植，雄性骨髓植入雄性受体，分别于第5、10、20天处死雌鼠。以雄性性别决定基因sry作为细胞标记，用原位杂交和FISH作为检测方法对骨髓细胞的肝细胞转化进行分析。结果 PCR移植效果初步分析可见，R BMT组11例中有10例PCR阳性；2AAF PH BMT组11例中有7例阳性，2AAF B BMT组10例中有6例阳性。sry原位杂交染色发现，第5天各组雌性受体肝索中均未见sry阳性的肝细胞。第10天R BMT组可见1例sry阳性的细胞位于肝细胞索,FISH染色可见这一细胞白蛋白mRNA阳性。第20天各组PCR阳性各例均可在肝索中检测到sry阳性的细胞。FISH染色可见白蛋白mRNA阳性。经统计学分析第20天各组sry阳性细胞数无明显差异。结论 在B BMT、2—AAF PH BMT和2—AAF R BMT模型中移植的骨髓细胞均可以植入肝脏，并存在于肝细胞索。植入肝索的骨髓细胞最早可见于移植后第10天，并发生转分化，表达白蛋白mRNA。不经过全身照射的2—AAF PH BMT组，移植的骨髓细胞也可以进入肝脏发生转分化，因此全身照射并不一定是移植骨髓细胞活化、植入和转化的必须条件。

Study on differentiation of bone marrow cells into hepatic lineages in rat

Objective To elucidate the possibility of transdifferention of bone marrow cell into liver cell rat. Methods Forty-five SD rats were randomly divided into three groups. 15 rats each. 1 . R + BMT (whole body irradiation and bone marrow transplantation); 2. 2-AAF + R + BMT; 3.2-AAF+ BMT + PH (75% hepatectomy) . The cross-sex BMT was performed and the rats were sacrificed on 5,10,20 day after bone marrow transplantion,sry gene positive cells in the liver of recipients, which derived from donor, were detected by in situ hybridization and fluorescence in situ hybridization (FISH). Results It was demonstrated that 10 of 11 cases were sry-positive in the R + BMT group,so did 7 of 11 cases in the 2-AAF + R + BMT group and 6 of 10 cases in the 2-AAF + PH + BMT group by PCR amplification. There was no sry-positive signal in hepatic plates by in situ hybridization at day 5 after BMT in all 3 groups although the PCR products were acquired in most rats sacrificed at day 5 after BMT. The sry-positive donor cells located in hepatic cord, occurred in one case in the R+BMT group at day 10 after BMT and expressed albumin mRNA detected by FISH staining. There were sry-positive cells located in hepatic cord in all PCR-positive cases at day 20 after BMT, and albumin mRNA expression showed by FISH staining. The numbers of sry-positive donor cells among three groups at day 20 after BMT were not statistically significant. Conclusion In all R + BMT,2-AAF + PH + BMT and 2-AAF + R + BMT models, transplanted bone marrow cells could engraft into liver and located in hepatic cord. Early in day 10 after BMT.it occurred and the engrafted cell could express albumin mRNA. Without whole body irradiation, the transplanted bone marrow cells could also engraft into liver and express albumin mRNA. So the whole body irradiation is optional to the activation,engraftment and differentiation of transplanted bone marrow cells.