内皮祖细胞和肝干细胞联合移植对大鼠肝纤维化的逆转作用

摘要

背景:目前骨髓源性内皮祖细胞(bone marrow-derived endothelial progenitor cells,BM-EPCs)或骨髓源性肝干细胞(bone marrow-derived hepatocyte stem cells,BDHSCs)移植治疗肝纤维化的研究较多见,但二者联合移植治疗肝纤维化的报道却罕见.因此设想将具有血管形成作用的 BM-EPCs 和具有肝细胞再生作用的BDHSCs联合移植,可发挥双重抗纤维化作用.目的:探讨BM-EPCs和BDHSCs联合移植对大鼠肝纤维化的逆转作用.方法:利用四氯化碳皮下注射6周建立大鼠肝纤维化模型.通过体外诱导培养获得肝纤维化大鼠来源的BM-EPCs,通过磁珠分选获得肝纤维化大鼠来源的BDHSCs.将两种细胞经尾静脉、门静脉分支输注移植到肝纤维化大鼠体内,观察其对肝纤维化程度和肝脏功能的改善作用.结果与结论:①Masson染色结果显示,单纯BDHSCs或BM-EPCs移植,以及两者联合移植均能抑制胶原纤维的形成.但肝纤维化分期评分结果显示,仅BM-EPCs/BDHSCs联合移植能明显改善肝纤维化程度,与模型组相比差异有显著性意义(P < 0.05);②肝功能和凝血功能检测结果显示,与模型组相比, BM-EPCs/BDHSCs 联合移植组谷丙转氨酶、谷草转氨酶、总胆红素、凝血酶原时间和部分活化凝血活酶时间5项指标水平显著改善(P < 0.05),与正常组水平相当;③结果表明,来源于肝纤维化大鼠的BM-EPCs/BDHSCs联合移植治疗肝纤维化有显著疗效,优于单一类型细胞移植.%BACKGROUND: At present, the transplantation of bone marrow-derived endothelial progenitor cells (BM-EPCs) or bone marrow-derived hepatocyte stem cells (BDHSCs) is common in the treatment of liver fibrosis, but the combined treatment for liver fibrosis is rarely reported. Combined transplantation of BM-EPCs possessing the function of angiogenesis and BDHSCs possessing the function of hepatocyte regeneration might play a dual anti-fibrosis role. OBJECTIVE: To evaluate the reversal effect on liver fibrosis by the combined transplantation of BM-EPCs and BDHSCs in rats. METHODS: The liver fibrosis rat models were induced with CCl4 subcutaneous injections for 6 weeks. BM-EPCs of rats with liver fibrosis were obtained by culture induction in vitro.BDHSCs of rats with liver fibrosis were obtained by magnetic bead cell sorting.BM-EPCs and/or BDHSCs were transplanted into liver fibrosis rats via the tail vein and branch of the portal vein,and then the effects of BDHSCs transplantatiron on liver fibrosis and liver function were observed. RESULTS AND CONCLUSION: (1) Masson staining results showed transplantations of BDHSCs and BM-EPCs, alone or both, could suppress the formation of collagen fibers. However, the staging scores of liver fibrosis showed that only the combined transplantation of BM-EPCs and BDHSCs could significantly improve liver fibrosis,which was significantly different from the model group(1.75±0.25 vs. 3.00±0.19, P < 0.05). (2) The liver biochemical assay in the blood showed that the levels of all five parameters of alanine aminotransferase, aspartate aminotransferase, total bilirubin, prothrombin time, and activated partial thromboplastin time in the BM-EPCs/BDHSCs group were significantly improved to be equivalent to normal levels, compared with those in the model group (P < 0.05). To conclude, it is an effective treatment for liver fibrosis by the co-transplantation of BM-EPCs and BDHSCs.