来源于肝纤维化环境内皮祖细胞移植治疗肝纤维化模型大鼠的逆转作用

Reversal effect of transplantation of bone marrow-derived endothelial progenitor cells from the liver fibrosis environment in rats with liver fibrosis

背景:理论上,从肝纤维化大鼠骨髓中提取经过体内病理环境"筛选"作用后的内皮祖细胞,应更能适应肝纤维化微环境,移植入纤维化肝脏后应更倾向于分化为成熟内皮细胞,参与肝内血管重建。目的:探讨来源于大鼠肝纤维化体内环境的骨髓源性内皮祖细胞移植对大鼠肝纤维化的治疗作用。方法:将28只Wistar大鼠随机分为3组:正常组(n=8)皮下注射橄榄油,2次/周;模型组(n=10)皮下注射四氯化碳3 m L/kg,2次/周,首剂加倍,分别于第2,3,5周末尾静脉输注生理盐水;实验组(n=10)皮下注射四氯化碳3 m L/kg,2次/周,首剂加倍,分别于第2,3,5周末尾静脉输注肝纤维化大鼠骨髓源性内皮祖细胞悬液。注射6周末,进行肝功能、凝血功能、肝组织相关蛋白及肝组织病理检查。结果与结论:(1)肝组织病理:模型组肝组织呈广泛而严重的肝细胞脂肪变性和坏死,门静脉和中央静脉周围炎症细胞浸润,汇管区扩大及大量纤维组织增生;与模型组相比,实验组肝细胞变性坏死、炎症细胞浸润及纤维组织增生等均有所减轻;(2)肝组织相关蛋白:与正常组相比,模型组透明质酸、层粘连蛋白和Ⅲ型前胶原、血管内皮生长因子、表皮生长因子水平升高(P<0.05);与模型组相比,实验组透明质酸、层粘连蛋白和Ⅲ型前胶原水平明显下降(P<0.05),血管内皮生长因子、表皮生长因子水平升高(P<0.05);(3)肝功能、凝血功能:与正常组相比,模型组肝功能、凝血功能严重受损(P<0.05);与模型组相比,实验组肝功能明显改善(P<0.05);(4)结果表明:来源于肝纤维化体内环境的骨髓源性内皮祖细胞移植能有效治疗大鼠肝纤维化,其作用机制可能是通过促进肝脏血管新生来实现的。

BACKGROUND：Theoretically, bone marrow-derived endothelial progenitor cells （EPCs） from liver fibrosis rats could be ＆quot;filtered＆quot; by the pathological environment in vivo. These EPCs would be more adapted to the micro-environment of liver fibrosis, and easier to differentiate into mature endothelial cells participating in the intrahepatic vascular remodeling after transplanted into the liver.OBJECTIVE：To explore the effectiveness of transplantation of bone marrow-derived EPCs from the liver fibrosis environment in liver fibrosis rats.METHODS：Twenty-eight Wistar rats were randomly divided into three groups as follows：normal group （n=8） were injected with olive oil, twice per week; model group （n=10） were infused with carbon tetrachloride at a dose of 3 mL/kg body weight （double doses for the first time）, twice per week, and infused with normal saline through the tail vein at 2, 3 and 5 weeks; EPCs transplantation group （n=10） were infused with carbon tetrachloride at a dose of 3 mL/kg body weight （double doses for the first time）, twice per week, and infused with EPCs suspension through the tail vein at 2, 3 and 5 weeks. Six weeks after final injection, the angiogenesis, hepatocyte proliferation and pathological changes in the liver tissues were observed. The liver function and coagulation function were tested.RESULTS AND CONCLUSION：（1） The pathological changes of the liver：in the model group, fatty degeneration and hepatocyte necrosis in the liver tissue were serious, inflammatory cells were infiltrated around the portal and central vein,the portal areas expanded, and fibrous tissues overgrew. Compared with the model group, these changes were significantly relieved in the EPCs transplantation group （P 〈 0.05）. （2） The expressions of liver-related proteins：compared with the normal group, the levels of hyaluronic acid, laminin, type III procollagen, vascular endothelial growth factor, epidermal growth factor were significantly increased in the model group （P 〈 0.05）. Compared with the model group, the levels of hyaluronic acid, laminin and type III procollagen were decreased significantly （P 〈 0.05）, and the levels of vascular endothelial growth factor and epidermal growth factor were increased in the EPCs transplantation group （P 〈 0.05）. （3） Liver function and coagulation function：compared with the normal group, the liver function and blood blotting function of rats were seriously damaged in the model group （P 〈 0.05）. Compared with the model group,the liver function and coagulation function were obviously improved in the EPCs transplantation group （P 〈 0.05）. To conclude, transplantation of bone marrow-derived EPCs from the liver fibrosis environment is effective for liver fibrosis in rats. The mechanism may be associated with the promotion of angiogenesis in the liver.