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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,雄性骨髓植入雌性受体,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,2AAM+R+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。FICH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R+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。
Objective To explore the feasibility of transforming myeloid cells into hepatocytes in vivo. Methods Female Sprague-Dawley rats were randomly divided into three groups of 15 rats each. ① R + BMT (whole body irradiation + bone marrow transplantation); ② 2-AAF + R + BMT: ③ 2-AAF + PH (partial hepatectomy) + BMT. Cross-sex bone marrow transplantation was performed. Female bone marrow was implanted into female recipients, and females were killed on the 5th, 10th and 20th day respectively. Male sperm was used to determine the gene sry as a cell marker, and in situ hybridization and FISH as a detection method were used to analyze the transformation of bone marrow cells into hepatocytes. Results Preliminary analysis of PCR results showed that 10 out of 11 patients in R + BMT group were positive for PCR, 7 in 11 patients in 2AAF + PH + BMT group and 6 in 10 patients in 2AAM + R + BMT group. sry in situ hybridization staining found that on the 5th day of each group of female recipient liver cord no sry positive hepatocytes. On the 10th day, one case of sry-positive cells was found on the hepatic cord in the R + BMT group. FISH staining showed that this cell albumin mRNA was positive. On the 20th day in each group PCR positive cases were detected in the liver sry-positive cells. FICH staining showed albumin mRNA was positive. By statistical analysis on the 20th day no significant difference in the number of sry-positive cells in each group. Conclusion Bone marrow cells transplanted in R + BMT, 2-AAF + PH + BMT and 2-AAF + R + BMT models can be implanted in liver and exist in hepatocellular cord. Bone marrow cells implanted hepatic cord can be seen on the first 10 days after transplantation, and transdifferentiation, expression of albumin mRNA. Transplanted bone marrow cells can also enter the liver to undergo transdifferentiation without systemic irradiation of 2-AAF + PH + BMT group. Therefore, systemic irradiation is not necessarily necessary for the activation, implantation and transformation of bone marrow cells.