NF-κB和P38MAPK在BMSCs肝向分化中的作用  

EFFECTS OF NF-κB AND P38MAPK IN HEPATIC DIFFERENTIATION OF MARROW MESENCHYMAL STEM CELLS

作　　者：王怡[1] 姜莎莎[1] 杨童茜 荆昭[1] 刘晓萍[1] 狄元璞 

机构地区：[1]青岛大学医学院组织学与胚胎学教研室,山东青岛266021 [2]美国匹兹堡大学环境与职业健康学院

出　　处：《青岛大学医学院学报》2017年第2期181-185,共5页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省自然科学基金项目(ZR2012CM008)

摘　　要：目的探讨信号转导分子核因子κB(NF-κB)和P38MAPK在肝细胞生长因子(HGF)、肝组织液(LTF)介导下骨髓间充质干细胞(BMSCs)向肝样细胞定向分化过程的作用。方法采用全骨髓贴壁法分离大鼠BMSCs,利用HGF、LTF两种诱导剂以及NF-κB信号通路抑制剂BAY 11-7082对第3代BMSCs进行分组培养。采用吲哚靛青绿(ICG)摄取实验检测肝向分化的细胞,免疫组化染色检测NF-κB蛋白表达,Western Blot法检测NF-κB、p-P38和AAT表达。结果经HGF、LTF诱导20d后的细胞可摄取ICG。免疫组化检测显示,BMSCs经HGF、LTF两种诱导剂诱导后,细胞核内有NF-κB蛋白表达,添加抑制剂BAY 11-7082后细胞核内NF-κB蛋白表达减弱(F=572.2、280.9,P<0.01)。Western Blot检测显示,经HGF、LTF两种诱导剂诱导后,细胞内NF-κB和p-P38蛋白表达量显著增加,且细胞能够表达AAT蛋白,加入抑制剂BAY 11-7082后随着培养时间的延长,NF-κB、p-P38和AAT蛋白表达量均出现不同程度的降低(F=280.8~3 028.2,P<0.01)。结论 HGF、LTF均可诱导BMSCs向肝细胞分化,P38MAPK和NF-κB信号通路均参与了BMSCs的肝向分化,P38MAPK信号通路的调控主要出现在肝向分化的前期,NF-κB的抑制在前期对其无明显影响。Objective To investigate the effects of signal transducer molecules,P38 MAPK and nuclear factor-kappa B（NF-κB）,on the hepatic differentiation of bone marrow mesenchymal stem cell（BMSCs）into hepatocyte-like cells under the mediation of hepatocyte growth factor（HGF）and liver tissue fluid（LTF）. Methods BMSCs were isolated by whole bone marrow adherent culture.The third generation of BMSCs were cultured with HFG,LTF,or BAY 11-7082（an inhibitor of NF-κB）.Indocyanine green（ICG）uptake assay was used to detect hepatic differentiation cells.Immunohistochemical staining was applied to measure the protein expression of NF-κB.Furthermore,Western blot analysis was used to measure the protein expression of several markers,i.e.,AAT,p-P38,and NF-κB. Results ICG uptake assay showed that the cells treated with HGF or LTF for20 days could take in ICG.Immunocytochemistry revealed that after treatment with HGF or LTF,NF-κB was expressed in the nucleus of BMSCs.After the addition of inhibitor,BAY 11-7082,the expression of NF-κB in cell nucleus was reduced（F=572.2,280.9;P〈0.01）.Western blot results indicated that the expression of NF-κB and p-P38 in cells significantly increased after induction with HGF or LTF,and AAT protein could be expressed.After addition of the inhibitor,the NF-κB,p-P38,and AAT protein expression decreased to different degrees with incubation time（F=280.8-3028.2,P〈0.01）. Conclusion Both HGF and LTF can induce the differentiation of BMSCs into hepatocytes,where the NF-κB pathway and P38 MAPK pathway participate in.The regulation of P38 MAPK pathway mainly takes effect in the early stage of hepatic differentiation,but the inhibition of NF-κB has little influence in the early stage.

关 键 词：间质干细胞 骨髓 NF-ΚB P38丝裂原活化蛋白激酶类 肝细胞 细胞分化 

分 类 号：R575.3[医药卫生—消化系统]