摘要
目的观察Regucalcin（RGN）在草酸钙结石大鼠肾脏和肾小管上皮细胞中的表达特点，探讨其在草酸钙结石形成过程中的作用及机制。方法采用1%乙二醇＋1%氯化铵自由饮用的传统方法构建大鼠肾草酸钙结石模型，通过偏光显微镜观察大鼠肾组织的晶体，并使用Western blot、实时荧光定量聚合酶链反应（FQ-PCR）、免疫组织化学（IHC）法检测RGN在草酸钙结石模型大鼠中的表达变化。使用不同浓度的一水草酸钙晶体（COM）刺激大鼠肾小管上皮细胞NRK-52E，通过Western blot和FQ-PCR方法检测RGN的表达。结果偏光显微镜观察显示：造模组全部大鼠肾组织切片肾小管内均有草酸钙晶体沉积，而对照组大鼠肾脏未发现晶体。Western blot法检测大鼠肾皮质RGN的表达发现，造模组大鼠肾组织RGN表达明显低于正常对照组，其mRNA的相对表达量为6%（t＝21.459，P＝0.000）；IHC法也同样显示造模组大鼠肾皮质RGN的表达降低，而且以晶体黏附处的肾小管上皮细胞降低尤为明显。使用COM处理大鼠肾小管上皮细胞NRK-52E后，检测RGN的表达发现，在不同浓度（0、25、50、100 μg/cm2）COM晶体处理后，RGN的表达均明显降低。COM 100 μg/cm2组mRNA的相对表达量为47.4%（t＝4.285，P＝0.013）。结论RGN有可能参与了大鼠肾草酸钙结石的形成和进展的调控。
Objective To study the expression of regucalcin （RGN） in rat models of calcium oxalate stone and the influence of RGN in normal rat renal tubules epithelia treated with calcium oxalate monohydrate （COM）.MethodsCalcium oxalate （CaOx） stone rat model was built by a standard diet with 1% ethylene glycol （28 days） and 1% ammonium chloride （7 days） added to the drinking water. Polarizing microscopy was used to detect CaOx crystals in the renal tubules of CaOx stone rat kidney. The expression of RGN in CaOx stone rat kidney was determined by Western blotting, real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） and Immunohistochemistry （IHC）. Renal tubular epithelial cells of normal rats （NRK-52E） were cultured and exposed to different levels of COM crystal （0, 25, 50, 100 μg/cm2）. Then, determine the expression of RGN in those cells treated with COM crystal.Results The CaOx stone rat model was successfully built. A large number of COM crystals were found in CaOx stone rat kidney by polarizing microscopy. In CaOx crystal stone rat model, RGN was significantly down-regulated in renal tissue by Western blotting and IHC, especially in the areas surrounded by the CaOx crystals. Compared with the control group, the relative expression of RGN mRNA is 6% （t=21.459, P=0.000）. RGN was also down-regulated in epithelia of renal tubules （NRK-52E） after treated with COM crystal. The relative expression of RGN mRNA is 47.4% （t=4.285, P=0.013） treated by COM （100 μg/cm2）.
Conclusion Our findings revealed that RGN was involved in the pathogenesis of CaOx stone formation in rat models.
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第10期1640-1642,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81570633）
广州市教育局一般项目（1201410963）
广州市科技局一般项目（201607010359）
