骨髓间充质干细胞移植在急性肝衰竭中的应用效果及对IL-10水平的影响研究

Effect of bone marrow mesenchymal stem cell transplantation on acute liver failure and its effect on IL-10 level

目的:探讨骨髓间充质干细胞移植在急性肝衰竭中的应用效果及对IL-10水平的影响。方法:采用差速离心法完成骨髓间充质干细胞分离及培养。制备大鼠急性肝衰竭动物模型。建模成功后随机将大鼠分为模型对照组(n=15)和治疗组(n=15),比较各组肝功能、炎症因子水平及肝脏组织切片HE染色。结果:①电子显微镜结果显示:分离制备的细胞多呈梭形生长,有粗大的突起,且细胞培养后5 d可见细胞成纤维样生长并向瓶底克隆;传代后细胞呈长梭形、纺锤形,未见细胞分化;流式细胞仪结果表明:分离制备的细胞中CD29+占99.5%、CD44+占96.4%、CD90+占96.70%,而CD45+仅为0.60%,说明获得的细胞纯度较高,符合骨髓间充质干细胞特性;②治疗组移植后ALT、AST、TBIL水平均低于模型对照组(P<0.05);治疗组移植后ALB水平高于模型对照组(P<0.05),治疗组与模型对照组ALT、AST、TBIL水平均高于假手术组(P<0.05);③治疗组移植后炎症因子TNF-α、IL-6及IL-10水平均低于模型对照组(P<0.05);④假手术组未参与建模,HE染色下肝细胞分布均匀、饱满,未见炎症反应;假手术组HE染色下肝小叶结构严重破坏,肝细胞膨胀、肿大,可见大量炎症细胞浸润;治疗组HE染色下肝细胞受到破坏,存在部分炎症细胞聚集,肝细胞略微肿大。结论:骨髓间充质干细胞移植用于肝衰竭中能提高肝功能水平,能降低IL-10水平,减轻肝脏局部炎症细胞聚集,从而缓解肝脏组织病理损伤。

Objective:To investigate the effect of bone marrow mesenchymal stem cell transplantation on acute liver failure and its effect on IL-10 level.Methods:Using differential centrifugation to complete the isolation and culture of bone marrow mesenchymal stem cells.Preparation of animal model of acute liver failure in rats.After successful modeling,the rats were randomly divided into model control group(n=15)and treatment group(n=15).Liver function,inflammatory factor levels and HE staining of liver tissue sections were compared.Results:①Electron microscopy showed that the cells prepared by the separation were long spindle-shaped and had large protrusions.After 5 days of cell culture,the cells were fibroblast-like and cloned into the bottom of the bottle.After passage,the cells were spindle-shaped and spindle-shaped.No cell differentiation was observed.The results of flow cytometry showed that CD29+accounted for 99.5%,CD44+accounted for 96.4%,CD90+accounted for 96.70%,and CD45+only 0.60%,indicating that the obtained cells were of higher purity and conformed to the bone marrow that characteristics of mesenchymal stem cells.②The levels of ALT,AST and TBIL in the treatment group were lower than those in the model control group(P<0.05);the ALB level in the treatment group was higher than that in the model control group(P<0.05).③The levels of ALT,AST and TBIL in the model control group were higher than those in the prosthetic group(P<0.05).The levels of inflammatory factors TNF-α,IL-6 and IL-10 in the treatment group were lower than those in the model control group(P<0.05).The prosthetic surgery group did not participate in the modeling.The hepatocytes were evenly distributed and plump under HE staining.④No inflammatory reaction was observed.The hepatic lobular structure was severely damaged by HE staining in the prosthetic surgery group,and the hepatocytes expanded and swollen.Large,visible a large number of inflammatory cell infiltration.Liver cells are destroyed under the treatment group HE staining,there was some accumulation of inflammatory cells,liver cells slightly swollen.Conclusion:Bone marrow mesenchymal stem cell transplantation can improve liver function in liver failure,reduce interleukin-10(IL-10)level,reduce local inflammatory cell aggregation in liver,and relieve liver tissue pathological damage.