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目的探讨小鼠骨髓来源干细胞(bone marrow stem cells,BMSCs)在体内分化为肝细胞的可行性及BMSCs是否参与肝功能受损的修复。方法 30只雌性C57BL/6小鼠全身一次性接受10 Gy60Co射线照射后,立即接受同品系雄性小鼠骨髓干细胞移植。移植后21 d后以CCl4致骨髓嵌合小鼠肝功能受损,并于CCl4致伤后第2、3、4、7、14、21、28天取骨髓嵌合小鼠肝脏,冰冻切片行Y染色体FISH和白蛋白(albumin,ALB)免疫荧光双标染色法,检测BMSC阳性细胞在受体小鼠肝内的分化情况,ELISA法检测肝脏基质细胞衍生因子-1(stromal cell-derived factor1,SDF-1)及其趋化因子受体(CXCR-4)、肝细胞生长因子(hepatocyte growth factor,HGF)、干细胞因子(stem cell factor,SCF)的表达。结果伤后2、3、4、7、14、21、28 d骨髓嵌合小鼠肝内皆可见FISH和ALB共表达的双阳性细胞,计数Y染色体FISH与ALB双阳性的细胞数量为3.763%~5.578%。ELISA法检测相关干细胞因子发现,在肝损伤后同一批次各时相点肝脏组织匀浆标本中SDF-1、CXCR-4、SCF、HGF 4种因子均呈现增高趋势,并在肝损伤第21天达到高峰表现,与其他时相点相比显著增加(P<0.05),同FISH与ALB荧光双标记共表达计数结果一致。结论 BMSCs可通过直接分化为肝细胞及分泌相关细胞因子间接促进受体肝脏干细胞增殖参与受损肝脏的修复。
Objective To investigate the feasibility of bone marrow-derived stem cells (BMSCs) differentiating into hepatocytes in vivo and whether BMSCs participate in the repair of impaired liver function. Methods A total of 30 female C57BL / 6 mice received a single dose of 10 Gy of 60Co radioactive rays and then received bone marrow stem cell transplantation of the same strain of male mice. After 21 days after transplantation, the liver function of the chimeric mice induced by CCl4 was impaired. The liver of chimeric mice was harvested on the 2nd, 3rd, 4th, 7th, 14th, 21st and 28th day after CCl4 injury. Y chromosome FISH and albumin (albumin, ALB) were used to detect the differentiation of BMSC-positive cells in the liver of recipient mice. The levels of stromal cell-derived factor 1 (SDF-1) and its chemokine receptor (CXCR-4), hepatocyte growth factor (HGF) and stem cell factor (SCF) Results Double positive cells co-expressed by FISH and ALB were observed in the liver of chimeric mice 2, 3, 4, 7, 14, 21, and 28 days after injury. The number of double positive FISH and ALB counts on chromosome Y was 3.763% ~ 5.578%. ELISA detection of relevant stem cell factor found in liver injury after the same batch of liver tissue homogenate samples of SDF-1, CXCR-4, SCF, HGF four kinds of factors showed an upward trend, and in liver injury 21 (P <0.05), which was consistent with the results of FISH and ALB fluorescent double labeling co-expression. Conclusion BMSCs can indirectly promote the proliferation of recipient hepatic stem cells and participate in the repair of damaged liver by directly differentiating into hepatocytes and secrete related cytokines.