Cldn14基因敲除对大鼠草酸钙结石诱导形成的影响及其机制  

Effects of Cldn14 gene knockout on the formation of calcium oxalate stones in rats and its mechanism

作　　者：罗培跃 郑丽英 陈涛 邹俊 李威 陈琪 程乐 甘立峰 张方涛 钱彪 LUO Peiyue;ZHENG Liying;CHEN Tao;ZOU Jun;LI Wei;CHEN Qi;CHENG Le;GAN Lifeng;ZHANG Fangtao;QIAN Biao(Department of Urology,The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000;Key Laboratory of Urology and Andrology of Ganzhou,Ganzhou 341000,China)

机构地区：[1]赣南医科大学第一附属医院泌尿外科,江西赣州341000 [2]赣州市泌尿男科重点实验室,江西赣州341000

出　　处：《现代泌尿外科杂志》2025年第2期168-173,共6页Journal of Modern Urology

基　　金：国家自然科学基金项目(No.82060135)。

摘　　要：目的探究紧密连接蛋白14(Cldn14)基因敲除对大鼠肾脏代谢及结石诱导形成的影响及其机制,为尿钙代谢及结石领域研究提供参考。方法将野生型大鼠与同周龄Cldn14基因敲除的纯合子大鼠分别随机分为野生型对照(WC)组、野生型乙二醇(WE)组、基因敲除对照(KC)组及基因敲除乙二醇(KE)组,每组10只。其中WE、KE组采用乙二醇+氯化铵方式诱导肾结石形成,WC和KC组采用生理盐水灌胃。标准维持饲料喂养4周后收集大鼠尿液,并进行静脉血检测,同时收集大鼠肾脏行HE、钙盐染色及透射电镜观察并提取肾组织蛋白。采用蛋白质印迹法检测紧密连接蛋白16(Claudin16)及紧密连接蛋白19(Claudin19)的表达水平。结果WE组与KE组肾小管腔内均可见晶体沉积,且KE组晶体多于WE组;相比于KC组,KE组在透射电镜下的肾小管上皮细胞中可见大量胞质内黑色结晶性包涵体,而WE组包涵体数量显著多于KE组。相比于WC和WE组,KC和KE组大鼠的血钙及血镁水平显著降低而尿钙水平则明显升高。此外,WE组相对于WC组、KE组相对于KC组尿钙水平进一步上升。KE组大鼠肾脏组织Claudin16水平显著降低,然而四组大鼠肾脏组织Claudin19水平差异均无统计学意义(P>0.05)。结论单纯敲除Cldn14基因并不能有效减少大鼠尿钙排泄及降低结石形成的风险,这可能与Claudin16水平降低有关。Objective To explore the effects of Cldn14 gene knockout on renal metabolism and stone formation in rats,so as to provide reference for research in the field of urinary calium metabolism and stone formation.Methods Cldn14 gene knockout homozygous rats and wild-type rats of the same age were randomly divided into 4 groups:wild-type control(WC)group,wild-type ethylene glycol(WE)group,gene knockout control(KC)group and gene knockout ethylene glycol(KE)group,with 10 rats in each group.The WE and KE groups were induced with ethylene glycol+ammonium chloride to form kidney stones,while the WC and KC groups received normal saline gavage.After 4 weeks of standard maintenance feeding,the urine samples were collected to detect the venous blood.The kidneys were collected for HE,Pizzolatto s staining and transmission electron microscopy.The protein in renal tissues was extracted to detect the expressions of Claudin16 and Claudin19.Results Crystal deposition was observed in the renal tubular lumen of the WE and the KE groups,and more crystals were detected in the KE group.The WE group had a large number of intracytoplasmic black crystalline inclusions observed in renal tubular epithelial cells under transmission electron microscope,followed by the KE and KC groups.Compared with WC and WE groups,KC and KE groups had significantly decreased serum calcium and magnesium levels but significantly increased urinary calcium level.In addition,the urinary calcium level was higher in the WE group than in the WC group and higher in the KE group than in the KC group.The KE group had lower level of Claudin16,but there was no significant difference in the level of Claudin19 among the 4 groups(P>0.05).Conclusion Knockout of Cldn14 gene alone cannot effectively reduce urinary calcium excretion or reduce the risk of stone formation in rats,which may be related to the decrease of Claudin16 level.

关 键 词：尿钙代谢 分子生物学 Cldn14基因 肾结石 紧密连接蛋白 乙二醇 

分 类 号：R691.4[医药卫生—泌尿科学]