骨髓间充质干细胞在肝细胞损伤恢复中的作用  

Role of bone marrow mesenchymalstem cells in recoveryprocess of hepatocyte injury

作　　者：徐土炳 李莉 罗兴迪 帅领[3] 赖洁娟[3] 

机构地区：[1]解放军第16医院普外科,新疆维吾尔自治区阿勒泰836500 [2]解放军第16医院医务处,新疆维吾尔自治区阿勒泰836500 [3]第三军医大学西南医院全军肝胆外科研究所,重庆市400038

出　　处：《实用医学杂志》2017年第5期687-692,共6页The Journal of Practical Medicine

基　　金：全军医学科技青年培育项目(编号:13QNP023)

摘　　要：目的:研究骨髓间充质干细胞(BMSCs)在肝细胞损伤恢复中的作用,并探讨可能机制。方法:(1)培养原代肝细胞及BMSCs并鉴定;(2)建立损伤肝细胞模型;(3)MTT法检测不同条件下肝细胞增殖情况;(4)加入BMSCs、bax抑制剂、BMSCs+bax抑制剂进行共培养,采用Western-blot和real-time PCR法检测TGF-β1、bcl-2、bax等指标。结果:(1)BMSCs+HGF共培养组中的OD值明显高于BMSCs或HGF单独联合培养组(P<0.05),BMSCs与HGF单独共培养组之间OD值差异无统计学意义,而它们与对照组相比差异均有统计学意义(P<0.05)。(2)bax1抑制剂共培养组与BMSCs共培养组的OD值差异无统计学意义,两者与对照组相比差异有统计学意义(P<0.05),BMSCs+Bax-1共培养组的OD值与BMSCs或bax1共培养组差异无统计学意义。(3)Western-blot法及real-time PCR检测结果提示TGF-β1、bax在bax1抑制剂共培养组与BMSCs共培养组中均显著低于对照组(P<0.05),而BMSCs+Bax-1抑制剂共培养组与两者单独共培养差异无统计学意义,Bcl-2结果与前两者相反。结论:BMSCs在肝细胞损伤恢复中能促进肝细胞增殖,BMSCs促进肝细胞增殖通过调节TGF-β1/bcl-2(bax)这个信号通道减少凋亡来实现的。Objective To study the role of bone marrow mesenchymalstem cells in recovery process of hepatocyteinjury and to explore the possible mechanism. Methods BMSCs and primary generation of hepatocytes were cultured and identified. A model of hepatocyte injure was established. Liver cell proliferation at different conditionswas detected by MTT method. Hepatocytes were added with BMSCs, baxinhibitor and mixture of BMSCs and baxinhibitor for co-cultured.Expressions of TGF-β1, bcl-2, and baxwere detected by Westernblot and real-time PCR. Results BMSCs and primary generation of hepatocyte were successfully cultured and identified, and a model of hepatocyte injury was successfully established. MTT tests revealed that the OD value of the mixture wassignificant higher in BMSCs and HGF co-culture group than in BMSCs co-culture group or HGF co-culture group ; there was no significant difference between BMSCs co-cuhure group and HGF co-culture group, while both groups were significant higher than control group. The MTF tests also revealed that the OD value of the mixture of BMSCs and Baxinhibitor co-culture group had no significant difference as compared with BMSCs co-cuhure group or Baxinhibitor co-culture group. There was no significant difference between BMSCs co-culture group and Baxinhibitor co-culture group, while the OD value was significant higher in both groups than in the control group. Westernb lot and real-time PCR results revealed that the value of TGF-β1 and bax in the mixture of BMSCs and Baxinhibitor co-culture group had no significant difference as compared with BMSCs co-culture group or Baxinhibitor co-culture group. There was no significant difference between BMSCs co-culture group and Baxinhibitor co-culture group, while both groups had a significant higher value than the control group. The value of Bcl-2 in the mixture of BMSCs and Baxinhibitor co-culture group had no significant difference as compared with BMSCs co-culture group or Baxinhibitor co-culture group. There was no significant difference between

关 键 词：骨髓间充质干细胞 TGF-Β1 BCL-2 BAX 实时荧光定量PCR 

分 类 号：R657.3[医药卫生—外科学]