尿酸性肾病小鼠模型的构建

CONSTRUCTION OF A MOUSE MODEL OF URATE NEPHROPATHY

目的构建尿酸性肾病小鼠模型。方法选取8周龄雄性尿酸氧化酶基因敲除小鼠16只(A组),8周龄雄性同窝野生型小鼠16只(B组),检测并比较两组小鼠血尿酸和血肌酐水平。随机取A、B组小鼠各4只,处死后取小鼠单侧肾脏,通过苏木精-伊红、Masson、免疫组化染色观察肾组织病理改变、纤维化程度及炎症状态,取另一侧肾脏通过偏振光显微镜观察尿酸盐结晶沉积情况。随机取A组小鼠6只,给予非布司他(8 mg/kg)灌胃降尿酸4周后,再次检测小鼠血尿酸、血肌酐水平,并观察肾脏尿酸盐结晶情况;同时取A、B组小鼠各6只,给予生理盐水灌胃4周作为对照。结果A组小鼠的血尿酸和血肌酐水平均显著高于B组小鼠(t=32.80、17.10,P<0.01)。A组小鼠肾脏有大量尿酸盐结晶沉积,存在明显肾盂积水、鲍曼氏囊和肾小管扩张、肾间质纤维化和大量炎性细胞浸润;B组小鼠肾脏未见明显病理改变。与生理盐水灌胃的A、B组小鼠相比,经非布司他治疗后的A组小鼠血尿酸和血肌酐水平明显下降(F=195.54、42.20,P<0.01)。经过4周降尿酸治疗后A组小鼠肾脏尿酸盐结晶沉积明显减少。结论尿酸氧化酶基因敲除的小鼠肾功能及肾脏组织受损,经非布司他治疗可改善其肾功能。该尿酸性肾病小鼠模型构建成功,可用于尿酸性肾病相关研究。

Objective To construct a mouse model of urate nephropathy.Methods A total of 16 male urate oxidase gene-knockout mice with an age of 8 weeks were selected as group A,and 16 male wild-type littermates aged 8 weeks were selected as group B.The levels of serum uric acid and serum creatinine were measured and compared between the two groups.Four mice each were randomly selected from groups A and B;the kidney at one side was collected after the mice were sacrificed to observe renal histopathological changes by HE staining,Masson staining,and immunohistochemical staining,and the kidney at the other side was collected to observe the deposition of urate crystal under a polarization microscope.Six mice were randomly selected from group A and were given febuxostat(8 mg/kg)by gavage for 4 weeks,and then the levels of serum uric acid and serum creatinine were measured and the deposition of urate crystal in the kidney was observed;6 mice each were randomly selected from groups A and B as controls.Results Group A had significantly higher levels of serum uric acid and serum creatinine than group B(t=32.80,17.10,P<0.01).Pathological results showed that the mice in group A had the deposition of a large amount of urate crystal in the kidney,with obvious hydronephrosis,dilatation of Bowman’s spaces and tubules,renal interstitial fibrosis,and inflammatory cell infiltration,while no obvious renal pathological changes were observed in group B.Compared with the mice from groups A and B that were given normal saline by gavage,the mice from group A had significant reductions in serum uric acid and serum creatinine after febuxostat treatment(F=195.54,42.20,P<0.01).In addition,group A had a significant reduction in the deposition of urate crystal in kidney after urate-lowering treatment for 4 weeks.Conclusion Damage of renal function and renal tissue is observed in urate oxidase gene-knockout mice,and an improvement in renal function is observed after febuxostat treatment.The mouse model of urate nephropathy is successfully constructed and can be used for the research on urate nephropathy.