尿毒症ApoE基因敲除小鼠加速发展的动脉粥样硬化与体内T细胞(Treg/Teff)平衡的关系  

Relationship between accelerated artherosclerosis and Treg/Teff balance in uremic apoE-/mice

作　　者：申燕[1] 袁祖贻[2] 刘艳[2] 肖嫣 吴岳[2] 赵燕[2] 田雨灵[2] 刘卫民[2] 王丽君[2] 梁萧[2] 陈涛[2] 耿涛[4] 

机构地区：[1]西安交通大学医学院第一附属医院肾内科,陕西西安710061 [2]西安交通大学医学院第一附属医院心内科,陕西西安710061 [3]北京市西山医院心内科,北京100144 [4]西安交通大学环境与疾病相关基因教育部重点实验室,陕西西安710061

出　　处：《南方医科大学学报》2010年第2期214-218,223,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(30871043;30971219);陕西省卫生厅科学研究基金(08D26)

摘　　要：目的建立尿毒症apoE-/-小鼠的动物模型,探讨模型鼠主动脉加速发展的动脉粥样硬化与体内T细胞(Treg/Teff)平衡的关系。方法选用C57BL/6J遗传背景的apoE-/-鼠为研究对象,通过右肾皮质电凝加左肾切除术建立尿毒症apoE-/-鼠模型,对照apoE-/鼠行假手术。造模后2周检测肾功能判断造模是否成功。造模后10周采血检测肾功、血清总胆固醇;应用ELISA法检测血清TGF-β1及IFN-γ浓度;应用流式细胞术检测脾脏CD4+CD25+Foxp3+Treg细胞比率;应用实时荧光定量PCR法检测主动脉组织中Foxp3及IFN-γ mRNA的表达;收集主动脉根部行冰冻切片油红O染色计算斑块相对面积。分析肾功能指标与Treg数量的相关性。结果造模后肾功能检测显示造模成功。造模后10周与对照组apoE-/鼠相比:尿毒症apoE-/-鼠主动脉根部动脉粥样硬化斑块相对面积显著提高,脾脏Treg细胞比率下降,血清TGF-β1浓度降低及IFN-γ、总胆固醇浓度升高,主动脉Foxp3 mRNA表达下调及IFN-γ mRNA表达上调。肾功能指标与Treg数量呈显著负相关。结论成功地建立了尿毒症apoE-/-小鼠模型。模型鼠主动脉加速发展的动脉粥样硬化与体内Treg数量减少、功能受损及Teff功能上调,即T细胞亚群(Treg/Teff)失衡有关。Objective To establish a uremic apolipoprotein E knockout（apoE-/-） mouse model and explore the relationship between accelerated atherosclerosis and Treg/Teff balance.Methods Using apoE-/-mice with C57BL/6J background,uremic apoE-/-mice were created by electrocautery of the right kidney and nephrectomy of the left,and the control apoE-/-mice received a sham-operation.Two weeks after inducing uremia,the renal function of the mice were evaluated to assess the validity of the model.Ten weeks after the operation,blood samples were obtained from the mice to assess the renal function and serum total cholesterol（TCH）;the serum concentrations of transforming growth factor-β1（TGF-β1） and interferon-γ（IFN-γ） were detected by ELISA,and CD4＋CD25＋Foxp3＋Treg ratio in the spleen was determined by flow cytometry.RT-PCR was used to detect the expression of Foxp3 and IFN-γ mRNA in the aorta,and oil red O staining used to investigate the relative atherosclerotic area on the frozen sections of the aortic root.The correlation between the renal function parameters and Treg quantity was analyzed.Results Renal function detection confirmed successful establishment of the uremic apoE-/-mouse model.Ten weeks after the operation,the relative atherosclerotic plaque area in the aortic root plaque increased significantly,the spleen Treg ratio decreased,the serum concentrations of TGF-β1 decreased and IFN-γ and TCH increased,the expression of aortic Foxp3 mRNA decreased and IFN-γ mRNA increased as compared with those in the control apoE-/-mice.A significant inverse correlation was found between the renal function parameters and Treg quantity in uremic apoE-/-mice.Conclusion In uremic apoE-/-mice,accelerated aortic atherosclerosis is correlated to the T cell subset（Treg/Teff） imbalance shown by decreased quantity and impaired function of Treg and enhanced activity of Teff.

关 键 词：尿毒症 ApoE-/-鼠 动脉粥样硬化 调节性T细胞 

分 类 号：R692.5[医药卫生—泌尿科学] R543.5[医药卫生—外科学]