尿酸盐在慢性肾病并发肾间质纤维化中的作用及机制  

The role and mechanism of urate in the development of interstitial fibrosis in chronic kidney disease

作　　者：杨萍 徐德苹 童子文 陈琼[1,3] 徐如月 周海胜 Yang Ping;Xu Deping;Tong Ziwen;Chen Qiong;Xu Ruyue;Zhou Haisheng(Dept of Biochemistry,Anhui Medical University,Hefei 230032;Dept of Clinical Laboratory,The Affiliated Hefei Hospital of Anhui Medical University,Hefei 230011;Dept of Clinical Laboratory,The Second People′s Hospital of Anhui Province,Hefei 230041;Center Scientific Research,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学生物化学教研室,合肥230032 [2]安徽医科大学附属合肥医院检验科,合肥230011 [3]安徽省第二人民医院检验科,合肥230041 [4]安徽医科大学科研实验中心,合肥230032

出　　处：《安徽医科大学学报》2024年第7期1206-1212,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82071832);安徽医科大学研究生科研与实践创新项目(编号:YJS20230003)。

摘　　要：目的探讨尿酸盐在慢性肾病并发肾间质纤维化过程中的作用及机制。方法利用0.2%腺嘌呤的饲料喂养小鼠9周,构建慢性肾病小鼠模型。造模结束后,小鼠眼眶后静脉丛采血,检测肾功能和血尿酸含量;HE染色和PAS染色分析肾脏组织病理变化;Masson染色观察肾脏纤维化程度;尿酸盐染色观察肾组织中尿酸盐沉积;Western blot和免疫组化检测目标分子的表达变化。利用不同浓度尿酸刺激原代培养小鼠肾小管上皮细胞(mRTECs)及人肾小管上皮细胞系(HKC);划痕实验观察尿酸对细胞迁移的影响。结果动物水平上,生化分析检测结果显示,模型组小鼠血清尿素氮(P=0.0064)、肌酐(P=0.0080)、血尿酸(P=0.0007)水平较对照组明显增加;HE和PAS染色结果显示,模型组小鼠出现严重肾小管损伤及炎症细胞浸润;Masson染色结果显示,模型组胶原沉积明显增加;尿酸盐染色结果显示,与对照组比较,模型组小鼠肾组织出现大量尿酸盐结晶;Western blot和免疫组化结果显示,模型组小鼠波形蛋白、平滑肌肌动蛋白和转化生长因子β1(TGF-β1)水平较对照组升高,E钙黏蛋白水平较对照组降低。细胞水平上,划痕结果显示,尿酸刺激促进肾小管上皮细胞迁移;Western blot检测结果与免疫组化一致。结论尿酸可通过促进肾小管上皮细胞分泌TGF-β1,进而促进上皮-间质细胞转分化,加速肾间质纤维化的发生。Objective To investigate the role and mechanism of urate in chronic kidney disease complicated with renal interstitial fibrosis(CKD-RIF).Methods Mice were continuously fed with a diet containing 0.2%adenine for a duration of 9 weeks to establish mice models with CKD-RIF.By the end of the 9-week experimental periods,collected blood samples from the posterior orbital venous plexus of mice to measure renal functions and serum urate concentrations prior to euthanizing the mice.Hematoxylin-eosin(HE)staining and periodic acid-Schiff staining(PAS)were used to investigate the pathological alternations in kidney tissues.Masson′s trichrome staining was used to observe the extent of renal fibrosis.Urate staining was used to detect urate deposition in renal tissues.Western blot and immunohistochemistry were used to detect the expression of target molecules.Scratch tests were used to examine the migration abilities of cells treated with different concentrations of uric acid.Results The kidney function analysis showed that a significant increase in the levels of serum urea nitrogen(P=0.0064),creatinine(P=0.0080)and urate(P=0.0007)in the CKD-RIF mice compared with the normal control group.The results of HE staining and PAS staining showed a significance of renal tubule injury and infiltration of inflammatory cells in the model group.Masson′s trichrome staining showed that a marked increase in collagen deposition in the model group.The results of urate staining showed a significant presence of urate crystals in kidney tissue of the model group when compared to the control group.Animal tissue immunoblotting and immunohistochemistry analysis showed a significant increase in the expression levels of vimentin,α-SMA and TGF-β1 in the model group in comparison to the control group.Conversely,in the model group,E-cadherin levels exhibited a dramatic reduction compared to the control group.The findings from the scratching tests showed that uric acid significantly enhanced cell migration.Western blot analysis showed a dramatic incr

关 键 词：尿酸盐 高尿酸血症 慢性肾病 肾间质纤维化 上皮-间质细胞转分化 

分 类 号：R34[医药卫生—基础医学]