不同剂量氧嗪酸钾联合腺嘌呤诱导大鼠高尿酸肾损伤模型的建立与评价

Establishment and evaluation of hyperuricemic nephropathy model induced by different doses of potassium oxanate combined with adenine in rats

目的通过不同剂量氧嗪酸钾联合腺嘌呤诱导大鼠高尿酸血症,探索大鼠高尿酸血症模型的最佳条件,为高尿酸血症的治疗研究提供可靠的动物模型。方法将体重220~240 g成年雄性Sprague-Dawley大鼠56只,按照每组8只分为正常对照组及不同剂量氧嗪酸钾(1000、1500 mg/kg)联合腺嘌呤(0、50、100 mg/kg)模型组6个,连续灌胃5周,每周对大鼠进行尾静脉采血,检测血清中血尿酸、血肌酐和血尿素氮水平。6周后取材,观察大鼠肾脏的病理改变,肾组织炎症、纤维化相关因子的mRNA及蛋白表达水平变化。结果1500 mg/kg氧嗪酸钾联合100 mg/kg腺嘌呤组大鼠造模2周后开始死亡,第6周生存率为62.5%;其余各组大鼠生存率与正常对照组比较,差异无统计学意义(P>0.05)。与正常组相比,造模1周后,各模型组大鼠血尿酸水平均有显著升高(P<0.05),第6周停止造模后血尿酸基本恢复到造模前水平;各模型组动物血肌酐、血尿素氮水平在第5周均较正常组有所升高;与正常组相比,模型组大鼠肾组织的炎症和纤维化相关因子mRNA和蛋白表达水平均随氧嗪酸钾联合腺嘌呤的剂量增加而增加,其中氧嗪酸钾1000 mg/kg联合腺嘌呤100 mg/kg组及氧嗪酸钾1500mg/kg联合腺嘌呤50 mg/kg组与正常组的差异有统计学意义(P<0.05)。大鼠肾脏解剖及组织学检测结果显示,模型组随着氧嗪酸钾及腺嘌呤剂量的增加,各组均有不同程度的肾髓质白色沉积,肾小管间质纤维化,小管上皮细胞坏死增加,肾小球萎缩。50 mg/kg腺嘌呤联合1500 mg/kg氧嗪酸钾组大鼠肾组织炎症与纤维化相关基因和蛋白的表达水平相比正常组有所升高,且可见组织炎性细胞浸润和纤维化的发生,符合高尿酸血症所致肾损伤的临床表现。结论氧嗪酸钾(1500 mg/kg)联合腺嘌呤(50 mg/kg)连续灌胃5周能够建立稳定的大鼠高尿酸肾损伤模型。

Objective To explore the optimal conditions of rat model of hyperuricemia(HUA)induced by different doses of potassium oxanate(PO)combined with adenine,and to provide reference for the treatment of HUA.Methods Male Sprague-Dawley rats(220-240 g body weight)were divided into normal control group,potassium oxanate(1000,1500 mg/kg)and adenine(0,50,100 mg/kg)combined model groups,with 8 rats in each group.After5 weeks of intragastric administration,blood were collected from tail vein of rats every week,and serum uric acid,creatinine and blood urea nitrogen level were measured.At the 6 th week,the changes of the pathological characteristics,expression of inflammatory and fibrosis-related factors in the kidneys were observed.Results In the 1500 mg/kg potassium oxanate combined with 100 mg/kg adenine group,rats died after 2 weeks of molding,and the survival rate at the 6 th week was 62.5%;but there was no significant difference between the other groups and the normal control group in survival rate(P>0.05).Compared with the normal group,the level of serum uric acid in each model group increased significantly after 1 week of molding(P<0.05),but recovered to the pre-model level after stopping intragastric administration in week 6.After 5 weeks,in model groups the levels of serum creatinine and blood urea nitrogen were higher than those in the normal control group;and the inflammation and fibrosis-related factors mRNA and protein expression of kidney tissue in model groups increased with the increase of ademine dose,and there was a significant difference in the PO 1000 mg/kg with adenine 100 mg/kg group,PO 1500 mg/kg with Adenine 50 mg/kg group compared to the normal control group(P<0.05).The results of renal anatomy and histology testing in rats showed that with the increased of the dosage of PO and adenine in the model groups,the increase of white deposition of renal medulla,tubulointerstitial fibrosis,and tubular epithelial cell necrosis was found,and the glomerular atrophy aggravated.Compared with the indexes in the normal control group,the expression levels of inflammation and fibrosis related genes and proteins in the 50 mg/kg adenine combined with 1500 mg/kg PO group were higher,and inflammatory cell infiltration and fibrosis were observed,which was consistent with the clinical manifestation of hyperuricemia induced renal injury.Conclusion PO(1500 mg/kg)combined with adenine(50 mg/kg)can establish a stable hyperuricemic nephropathy model in rats.