高尿酸血症肾损害动物模型建立及发生机制研究  

Establishment of uric acid associated nephropathy animal model and the underlying mechanisms

作　　者：王莉[1] 徐柳青[1] 施映枫 熊重祥[1] 鲍文芳 严海东[1] 刘娜[1] 庄守纲[1] 

机构地区：[1]同济大学附属东方医院肾内科,上海200120

出　　处：《中华肾脏病杂志》2015年第3期203-207,共5页Chinese Journal of Nephrology

基　　金：上海市浦江人才项目（13PJ1406900）;国家自然基金项目（81200492,81470991,81470920,81270778,81170638）;浦东新区卫生系统重点学科建设基金资助（PWZx2014-06）

摘　　要：目的 建立高尿酸血症肾损害大鼠模型,研究高尿酸血症肾损害可能的发生机制.方法 选用SPF级雄性SD大鼠18只（6～8周龄）,体质量200～ 220 g,随机分入正常对照组和高尿酸血症肾损害模型组,每组9只.正常对照组大鼠早晚两次蒸馏水灌胃;模型组大鼠早晚两次腺嘌呤（0.1 g·kg-1·d-1）与氧嗪酸钾（1.5 g·kg-1·d-1）混悬液灌胃.连续给药21 d处死大鼠,留取血清检测尿酸（UA）、肌酐（Scr）,留取大鼠肾脏组织,行PAS及Masson染色;应用黄嘌呤氧化酶（XOD）试剂盒,检测大鼠黄嘌呤氧化酶活性变化;Western印迹检测大鼠磷酸化表皮生长因子（p-EGFR）、总表皮生长因子（EGFR）的表达.免疫组化检测大鼠α平滑肌肌动蛋白（α-SMA）的表达.结果 与正常对照组大鼠比较,高尿酸血症肾损害模型组大鼠血尿酸及Scr均显著增高;PAS染色可见肾小球硬化,肾小管上皮细胞胞浆空泡,肾小管排列不规则、肥大、管腔扩张,炎症细胞浸润;Masson染色可见肾间质纤维化面积显著增加;模型组XOD活性显著增高[（52.68±9.79） μmol/L比（32.23±6.72）μmol/L,P＜0.05];Western印迹显示模型组磷酸化EGFR表达上调;免疫组化显示模型组间质区域α-SMA表达显著上调.结论 腺嘌呤与氧嗪酸钾联合灌胃法成功建立高尿酸血症肾损害模型.该模型可能通过调控肾脏EGFR磷酸化,上调α平滑肌肌动蛋白,活化肾间质成纤维细胞介导肾脏损伤进展.Objective To establish an uric acid associated nephropathy （UAN） animal model and explore the mechanisms involved.Methods Eighteen （6-8 weeks old） male Sprague-Dawley rats weighed 200-220 g were randomly assigned into 2 groups：the control group （n=9）,the uric acid associated nephropathy group （n=9）.UAN rat model was established by oral administration of adenine （0.1g/kg） and potassium oxonate （1.5 g/kg） mixture daily for 3 weeks.After 3 weeks,the rats were sacrificed and blood and kidney samples were collected.Serum uric acid,creatinine and other biochemistry index were measured weekly.PAS and Masson staining were conducted to evaluate renal pathology and renal fibrosis.Serum activity of xanthine oxidase （XOD） was examined.Expression of p-EGFR and EGFR were detected by western blot and α-SMA expression was detected by immunohistochemical staining.Results After 3 weeks,the model group rats got 1.5 folds increased serum creatinine and significantly elevated serum uric acid.PAS and Masson staining showed that the UAN kidney developed glomerulosclerosis,tubulointerstitial damage and inflammatory cell infiltration.Serum activity of xanthine oxidase （XOD） was significantly upregulated in UAN group [（52.68±9.79） μ mol/L vs （32.23±6.72） μ mol/L,P 〈 0.05].Western blot showed that EGFR was activated and α-SMA expression increased remarkably in renal interstitial area of UAN rats.Conclusions The mixture of adenine and potassium oxonate can successfully establish UAN model.Phosphorylation of EGFR may mediate the activation of renal interstitial fibroblasts and accelerate the development and progression of UAN.

关 键 词：高尿酸血症 受体 表皮生长因子 肌动蛋白类 

分 类 号：R589.7[医药卫生—内分泌] R-332[医药卫生—内科学]