沉默钙敏感受体对大鼠尿钙排泄影响及对紧密连接蛋白14的作用  

Effect of silencing Ca SR on urinary calcium excretion in rats and its effect on CLDN14

作　　者：崔庆鹏 刘孝东[2] 范世成 朱元全 罗云[2] 罗钰辉[2] CUI Qingpeng;LIU Xiaodong;FAN Shicheng;ZHU Yuanquan;LUO Yun;LUO Yuhui(Department of Urology,the Third People's Hospital of Yunnan Province,Kunming 650000,China)

机构地区：[1]云南省第三人民医院泌尿外科,昆明650000 [2]昆明医科大学第一附属医院泌尿外科,昆明650032

出　　处：《实用医学杂志》2020年第14期1882-1885,1891,共5页The Journal of Practical Medicine

基　　金：云南省应用基础研究项目(昆医联合专项)(编号:2017FE648);昆明医科大学博士基金项目(编号:2015BS026);国家自然科学基金项目(编号:8186045)。

摘　　要：目的了解沉默大鼠钙敏感受体(calcium-sensing receptor,CaSR)基因后,肾脏组织紧密连接蛋白14(the tight junction protein claudin 14,CLDN14)的表达情况及24 h尿钙排泄量变化情况。方法 (1)建立遗传高钙尿结石形成(GHS)大鼠模型;(2)CaSR-miRNA干扰病毒分别干扰GHS组大鼠和正常尿钙对照组大鼠(NC)作为实验组,生理盐水干预作为对照组。(3)处死大鼠取双肾并测24 h尿钙;(4)RT-PCR和Western Blot技术检测大鼠肾上皮细胞CLDN14的mRNA和蛋白表达水平。结果 (1)CaSR-miRNA干扰病毒后,与对照组大鼠相比,GHS大鼠组尿钙排泄量减少,差异有统计学意义(P <0.05);NC组大鼠尿钙排泄量变化不明显,差异无统计学意义(P> 0.05)。(2)GHS组大鼠CLDN14 mRNA和蛋白表达量减少,差异有统计学意义(P <0.05);NC组大鼠CLDN14 mRNA和蛋白表达无明显变化,差异无统计学意义(P>0.05)。结论 (1)干扰病毒体内干扰GHS大鼠后尿钙排泄量下降,NC组大鼠无此现象。(2)CaSR可能通过调控CLDN14的表达来调节肾脏对Ca2+的重吸收。Objective To explore the expression of CLDN14 and 24-hour urinary calcium excretion in thekidney after silencing the calcium-sensing receptor(Ca SR)gene.MethodsEstablish the genetic hypercalciuricstones(GHS)forming rat models. Ca SR-mi RNA interference virus interfered with rats in GHS group and rats innormal urinary calcium control group(NC)were served as the experimental group,while rats with normal salineintervention were served as the control group. Both kidneys were taken and urinary calcium was measured for 24 hbefore the rats were sacrificed. RT-PCR and Western Blot technology to detect the m RNA and protein expressionlevels of CLDN14 in rat renal cells.Results(1)Compared with the control group,the urinary calcium excretionof rats in the GHS group was decreased after internal interference(P < 0.05). There was no significant change inurinary calcium excretion in the NC group(P > 0.05).(2)The m RNA and protein expression levels of CLDN14 inGHS group were decreased,and the difference was statistically significant(P < 0.05). There was no significantchange in CLDN14 m RNA and protein expression in the NC group(P > 0.05).Conclusions(1)The urinary calci-um excretion of GHS rats is decreased after the interference of virus in vivo,which is not found in the NC group.(2)Ca SR may regulate the reabsorption of Ca2+ in the kidney by regulating the expression of CLDN14.

关 键 词：钙敏感受体 泌尿系结石 特发性高钙尿症 GHS大鼠 体内RNA干扰 紧密连接 

分 类 号：R6965[医药卫生—泌尿科学]