TAK1信号通路在高糖诱导骨髓来源巨噬细胞激活中的作用  

Role of TAKI signaling pathways in bone marrow derived macrophage activation induced by high glucose

作　　者：冯世尧 徐兴欣[1] 邵云侠[1] 李媛媛[1] 付欣[1] 吴永贵[1] 

机构地区：[1]安徽医科大学第一附属医院肾内科,合肥230022

出　　处：《中华肾脏病杂志》2016年第1期37-42,共6页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81270813）

摘　　要：目的观察高糖对骨髓来源巨噬细胞（bonemal Tow-derive dmacrophages，BMDM）激活的影响，并探讨转化生长因子B激活激酶1（TGF-pactivatedkinase．1，TAK1）信号通路在其中的作用机制。方法分离获取小鼠BMDM，运用流式细胞术鉴定BMDM细胞纯度。高糖作为刺激因素，TAK1特异性抑制剂5z．7-oxozeaenol作为干预因素，分别设正常对照组（1640培养基）、渗透浓度对照组（25mmol／L甘露醇）、高糖组（33mmol／L葡萄糖）和高糖＋抑制剂组（33mmo]／L葡萄糖＋300nmol／L5Z-7-oxozeaen01）。采用细胞免疫荧光和流式细胞术检测BMDMM1表型，实时定量PCR检测各组细胞单核细胞趋化因子1（MCP-1）和肿瘤坏死因子α（TNF—n）mRNA水平，Western印迹法检测各组磷酸化（P）-TAKl、TAKl结合蛋白（TAKlbindingprotein，TABl）、p-JNK、p-p38MAPK和NF．KBp65蛋白的表达。结果BMDM诱导分化成功，其纯度达99．36％。10—300nmol／L5Z．7．oxozeaenol对高糖环境中BMDM细胞活性无影响（均P〉0．05）。与正常对照组比较，高糖组M1型巨噬细胞增加，MCP．1、TNF-αmRNA水平均上调（均P〈0．01），P-TAK1、TAB1、P—JNK、p-p38MAPK、NF-KBp65蛋白表达也显著升高（均P〈0．05）；TAKl特异性抑制剂5z．7-oxozeaenol作用均能抑制高糖诱导产生的效应（均P〈0．05）。结论高糖能诱导BMDM向M1表型转化，TAK1特异性抑制剂5-一7．oxozeaenol可能通过抑制TAKl／MAPKs和TAKl／NF-KB通路，抑制巨噬细胞M1型活化和炎性因子的表达。Objective To investigate the role of transforming growth factor-β activated kinase- 1 （TAK1） signaling pathway in the activation of bone marrow derived macrophages （BMDM） induced by high glucose. Methods Purity of mouse BMDM was detected by flow cytometry. The mice macrophages cultured in vitro were stimulated by high glucose and treated with TAKI specific inhibitor 5Z-7-oxozeaenol. Cells were divided into normal control group （RPMI 1640）, osmolality control group （25 mmol/L mannitol）, high glucose group （33 mmol/L D- glucose） and inhibitor group （33 mmol/L D-glucose＋300 nmol/L 5Z-7-oxozeaenol）. Immunocytochemistry and flow cytometry were used to detect macrophage subtype. The expression of monocyte chemotactic protein- 1 （MCP- 1） and tumor necrosis Factor-a （TNF-α） mRNA were determined by real time PCR. Expressions of p-TAK1, TAK1 binding protein （TAB1）, p-JNK, p-p38 MAPK and NF-KB p65 proteins were analyzed by Western blotting. Results The purity of BMDM was about 99.36%. Compared with normal control group, high glucose group had increased percentage of MI macrophages, increased expression of MCP-I and TNF-α mRNA （all P 〈 0.05）. Moreover, p-TAK1, TAB1, p-JNK, p-p38 MAPK and NF-KB p65 proteins expression also increased significantly in high glucose group （all P 〈 0.05）. After treatment with inhibitor 5Z- 7 - oxozeaenol, the effects induced by high glucose were inhibited （P 〈 0.05）. Conclusions High glucose can induce M1 macrophage activation and expression of inflammatory cytokine of BMDM, which can be inhibited 5Z-7-oxozeaenol through inhibiting TAK1/MAPK and TAK1/NF-KB pathway.

关 键 词：巨噬细胞活化 MAP激酶激酶激酶类 MAP激酶信号系统 炎症 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]