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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 an animal model of uremic apoE - / - mice and explore the relationship between the accelerated development of atherosclerosis and the balance of T cells (Treg / Teff) in vivo. Methods apoE - / - mice with genetic background of C57BL / 6J were selected as the study object. The uremic apoE - / - murine model was established by right renal cortex electrocoagulation combined with left nephrectomy. The control apoE- / mice were sham operated. 2 weeks after modeling to determine renal function to determine the success of modeling. Serum TGF-β1 and IFN-γ levels were measured by ELISA. The ratio of CD4 + CD25 + Foxp3 + Treg cells in spleen was detected by flow cytometry. Real-time fluorescence quantitative PCR Method was used to detect the expression of Foxp3 and IFN-γ mRNA in aorta tissue. Frozen sections of the aortic root were collected for oil red O staining to calculate the relative area of plaque. The correlation between renal function and Treg was analyzed. Results Renal function test showed that the model was successful. Compared with apoE / mice in control group, the relative area of atherosclerotic plaque in aortic root of apoE - / - mice increased significantly at 10 weeks after modeling, the ratio of Treg cells in spleen decreased, the concentration of serum TGF-β1 decreased and IFN -γ, elevated total cholesterol concentration, downregulation of aortic Foxp3 mRNA expression and up-regulation of IFN-γ mRNA expression. There was a significant negative correlation between renal function indexes and the number of Tregs. Conclusion ApoE - / - mouse model of uremia was successfully established. Atherosclerosis accelerated development of the aorta in model mice is associated with decreased Treg numbers, impaired function and upregulated Teff function, ie an imbalance of T cell subsets (Treg / Teff).