天津医药

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钙敏感受体及紧密连接蛋白-14在草酸钙结石大鼠肾组织中的表达及意义

孙稳1,2,王勤章2,丁国富1   

  1. 1. 石河子大学医学院
    2. 石河子大学医学院第一附属医院泌尿外科
  • 收稿日期:2013-11-05 修回日期:2014-03-11 出版日期:2014-07-15 发布日期:2014-07-15
  • 通讯作者: 王勤章

Expressions of CaSR and Claudin-14in Renal Calcium Oxalate Stone Model of Rats

  • Received:2013-11-05 Revised:2014-03-11 Published:2014-07-15 Online:2014-07-15

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摘要：

【摘要】目的 初步探讨钙敏感受体（CaSR）和紧密连接蛋白（Claudin）-14在肾草酸钙结石大鼠肾组织中的表达及意义。方法 30只雄性SD大鼠随机分为对照组和结石组各15只，采用1%乙二醇和2%氯化铵每日2mL灌胃诱导制作大鼠肾草酸钙结石模型。免疫组织化学检测CaSR蛋白表达；RT-PCR检测Claudin-14mRNA表达；West? ern-Blotting分别检测CaSR和Claudin-14蛋白表达；全自动生化分析仪检测2组大鼠肾功能、血及尿生化指标。结果 光镜下结石组有大量结石结晶形成，血清肌肝（Cr）、尿素氮（BUN）、24h尿钙及尿量较对照组升高（P＜0.01），2 组间血钙、尿pH差异无统计学意义。结石组Claudin-14mRNA及CaSR蛋白表达较对照组升高（P＜0.01），Claudin- 14蛋白在结石组大鼠肾组织中呈特异性高表达，对照组未检测到Claudin-14蛋白表达；结石组大鼠肾组织中CaSR 和Claudin-14蛋白表达呈正相关，同时，CaSR、Claudin-14蛋白表达与24h尿钙排泄量也呈正相关。结论 CaSR和 Claudin-14表达增高可通过增加尿钙排泄量，进而参与结石的形成。

关键词: 肾结石, 草酸钙, 受体, 钙敏感, 大鼠, Sprague-Dawley, 紧密连接蛋白-14

Abstract:

[Abstract] Objective To investigate the expressions of calcium sensitive receptor (CaSR) and tight junction protein (Claudin)-14in renal calcium oxalate stone rat model. Methods Thirty male SD rats were randomly divided into control group (n=15) and model group (n=15). The rat model of renal calcium oxalate stone was established by gavaging 1% glycol (2 mL/d) and2% ammonium chloride. The expression of CaSR protein was detected using immunohistochemical assay. RT PCR was used to detect the Claudin-14mRNA expression. The expression levels of CaSR and Claudin-14protein were de? tected by Western blot assay respectively. The full automatic biochemical analyzer was used to detect rat renal function and changes of blood and urine biochemical indices.Results A large stone crystallization was observed under light microscope in model group. The serum levels of Cr, BUN, 24-h urine calcium and urine volume were significantly higher in model group than those of control group (P＜0.01). There were no significant differences in serum calcium level and urinary pH value be? tween two groups. The expression levels of Claudin-14mRNA and CaSR protein were significantly higher in model group than those of control group (P＜0.01). The Claudin-14protein expression was specifically higher in renal tissues of model group. There was no Claudin-14protein expression in control group. There was a positive correlation between CaSR and Claudin-14protein expression in renal tissues of model group. Also, CaSR and Claudin-14protein expressions were posi? tively correlated with the 24-h urine volume.Conclusion The increased expressions of CaSR and Claudin-14involved in the formation of kidney stone by increasing the urinary calcium excretion.

Key words: kidney calculi, calcium oxalate, receptors, calcium-sensing, rats, Sprague-Dawley, claudin-14