摘要目的 研究钙结合蛋白Calbindin-D28k在遗传性高钙尿结石(GHS)大鼠肾脏中的表达,并探讨其在特发性高钙尿症发病中的作用.方法 分别取16只GHS大鼠与16只正常对照组(NC)大鼠新鲜肾脏组织,Western印迹检测Calbindin-D28k蛋白表达水平,实时荧光定量PCR检测其基因表达水平.结果 GHS大鼠Calbindin-D28k蛋白表达水平显著高于NC大鼠(4值分别为0.49±0.02和0.20±0.01,P＜0.05),GHS组Calbindin-D28k mRNA表达水平是NC组的1.21倍[2-(△△CT)值分别为1.21和1.0,P＞0.05].结论 Calbindin-D28k的表达上调可能是由GHS大鼠的维生素D受体高表达和高尿钙所引起,在尿钙重吸收中起重要作用,但不是导致特发性高钙尿症的关键分子.

abstractsObjective To study the expression level of calbindin-D28k,a kind of calcium binding protein,in the kidneys of genetic hypercalciuric stone-forming(GHS)rats and to investigate its role in idiopathic hypercalciuria(IH).Methods Kidneys were taken out from 16 GHS rats and 6 normal control (NC)rats.Westem blotting and real time quantitative PCR were used to detect the protein and mRNA expression levels of calbindin-D28k respectively.Resuits Western blotting showed that the A value of calbindin-D28k of the GHS rats was 0.49±0.02.significantly higher than that of the NC rats (0.20±0.01,P＜0.05).The 2-(△△CT)value of mRNA of calbindin-D28k of the GHS rats was 1.21,remarkably higher than that of the Nc rats[with the of 2-(△△CT)value of 1.0].There was not significant difference in tIle △CT value between the two groups(P＞0.05).Conclusion The up-regulation of calbindin-D28k in the GHS rats is possibly caused by hyperexpression of VDR and hypercalcinuria,and plays an important role in urine calcium reabsorption;however,it is not the key protein that results in IH.