Establishment and analysis of a rat model of chronic renal allograft dysfunction complicated with insulin resistance

doi:10.3969/j.issn.2095-4344.2017.12.016

Abstract

Abstract:

BACKGROUND: The main cause of chronic renal allograft dysfunction (CRAD) remains unclear until now, and insulin resistance (IR) may be a cause of CRAD. However, there is a lack of an available animal model for research on their association.
OBJECTIVE: To establish a rat model of CRAD complicated with IR, and to investigate the effect of IR on function, urinary protein and pathologic changes of the renal allograft.
METHODS: The high-fat diet was administrated to a F344-to-Lewis rat model of CRAD to induce IR. There were three groups including CRAD+IR, CRAD and control (uninephrectomy) groups, and the serum insulin, glucose, creatinine, urea nitrogen and 24-hour urinary protein levels in each group were measured and insulin resistance index was calculated at 8, 12, and 16 weeks after transplantation. Meanwhile, oral glucose tolerance and insulin tolerance tests were performed and the areas under glucose-time curve were calculated. The status and stability of CRAD+IR models were assessed based on all above indicators at each time point. The serum creatinine, urea nitrogen and 24-hour urinary protein levels were compared between CRAD+IR and CRAD groups. Pathological evaluation was conducted at 16 weeks based on the total Banff scores.
RESULTS AND CONCLUSION: The insulin resistance index, and area under glucose-time curve of the oral glucose tolerance and insulin tolerance tests in the CRAD+IR group were higher than those in the CRAD and control groups, and all above indexes showed no significant changes with time. The serum creatinine, urea nitrogen and 24-hour urinary protein levels in the CRAD+IR group were lower than those in the CRAD group, and showed no obvious changes with time. Slightly thickened basement membrane of glomeruli and tubules, interstitial fibrosis and mild arterial intimal hyperplasia were observed both in the CRAD+IR and CRAD groups, but the pathological degree did not differ notably between the two groups. These results indicate that the stable CRAD+IR models can be established via feeding CRAD rats with high-fat diet for 8 weeks. IR can cause glomerular hyperfiltration and albumin overload-induced tubular injury which may potentially affect the function and urinary protein level of the renal allograft.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Kidney Transplantation, Renal Insufficiency, Graft Survival, Insulin Resistance, Metabolic Syndrome X, Disease Models, Animal, Tissue Engineering

CLC Number:

R318

Lv Dao-yuan1, 2, Zhou Qin1, 2, Xia Yue1, 2, You Xu1, 2, 3, Zhao Zhi-hong4, Li Yong-qiang1, 2, Zou He-qun1, 2 . Establishment and analysis of a rat model of chronic renal allograft dysfunction complicated with insulin resistance[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(12): 1894-1902.

Figures/Tables 2

2.1 实验动物数量分析实验中31只大鼠造模成功，分为3组，即对照组(n=10)、慢性移植肾失功组(n=10)及胰岛素抵抗慢性移植肾失功组(n=11)，全部纳入结果分析，无脱失。 2.2 一般情况及体质量测定入组31只大鼠观察期间均无死亡，术后早期各组大鼠精神状态佳，活动良好，反应灵敏，毛发整齐有光泽，饮食及尿便正常。随观察时间延长，胰岛素抵抗慢性移植肾失功组较慢性移植肾失功组及对照组活动减少，反应尚可，毛发稍凌乱。如图1所示，观察期间各组大鼠体质量均稳步上升，胰岛素抵抗慢性移植肾失功组体质量上升速度高于慢性移植肾失功组及对照组，慢性移植肾失功组与对照组体质量上升速度接近。自术后第7-16周，3组间体质量的差异有显著性意义，胰岛素抵抗慢性移植肾失功组体质量大于慢性移植肾失功组及对照组，提示单纯高脂饲料喂养7周左右可使慢性移植肾失功大鼠发生明显肥胖。 2.3 生化检查 2.3.1 胰岛素抵抗评估如表1所示，术后8，12，16周，胰岛素抵抗慢性移植肾失功组胰岛素抵抗指数、口服葡萄糖耐量及胰岛素耐量试验血糖时间曲线下面积均大于慢性移植肾失功组及对照组。纵向比较胰岛素抵抗慢性移植肾失功组3个时间点胰岛素抵抗指数(F=0.129，P=0.880)、口服葡萄糖耐量试验血糖时间曲线下面积(F=0.501， P、胰岛素耐量试验血糖时间曲线下面积(F=0.661，P=0.524)差异均无显著性意义。提示胰岛素抵抗慢性移植肾失功模型已于8周成功建立并于8-16周保持稳定。=0.611) 2.3.2 血胰岛素、血糖及血脂检测如表2所示，术后8周，胰岛素抵抗慢性移植肾失功组血胰岛素、血糖、低密度脂蛋白均大于慢性移植肾失功组及对照组。而三酰甘油(F=1.636，P=0.213)、总胆固醇(F=2.703，P=0.084)、高密度脂蛋白(F=0.925，P=0.408)3组间差异均无显著性意义。术后12，16周，胰岛素抵抗慢性移植肾失功组血胰岛素、血糖亦均大于慢性移植肾失功组及对照组；总胆固醇、低密度脂蛋白大于对照组，而与慢性移植肾失功组差异无显著性意义；三酰甘油(F12周=0.278，P12周=0.760；F16周= 1.045，P16周=0.365)、高密度脂蛋白(F12周=0.538，P12周= 0.590；F16周=0.406，P16周=0.670)3组间差异均无显著性意义。纵向比较3个时间点胰岛素抵抗慢性移植肾失功组上述指标差异，仅16周低密度脂蛋白大于8周及12周，其余血胰岛素(F=0.070，P=0.933)、血糖(F=0.196，P=0.823)、三酰甘油(F=0.36，P=0.700)、总胆固醇(F=2.774，P=0.078)、高密度脂蛋白(F=0.145，P=0.866)3个时间点差异均无显著性意义。提示单纯高脂饲料喂养显著且稳定地升高了慢性移植肾失功大鼠血胰岛素及血糖，对其血脂有提升趋势，但尚不显著。 2.3.3 肾功能及尿蛋白检测如表3所示，术后8，12，16周，胰岛素抵抗慢性移植肾失功组血肌酐、尿素氮均小于慢性移植肾失功组，慢性移植肾失功组血肌酐、尿素氮均大于对照组，纵向比较胰岛素抵抗慢性移植肾失功组及慢性移植肾失功组3个时间点血肌酐(F胰岛素抵抗慢性移植肾失功组= 0.978， P胰岛素抵抗慢性移植肾失功组=0.388；F慢性移植肾失功组=2.675， P慢性移植肾失功组=0.087)、尿素氮(F胰岛素抵抗慢性移植肾失功组=1.115， P胰岛素抵抗慢性移植肾失功组=0.341；F慢性移植肾失功组=1.714， P慢性移植肾失功组=0.199)差异均无显著性意义。提示慢性移植肾失功组肾功能低于对照组且于观察时间内保持稳定，而胰岛素抵抗可能稳定地影响移植肾功能。"

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