目的对草酸钙尿石症患者肾脏组织中维生素K环氧化物还原酶亚单位1(VKORCl)剪接异构体进行分析。方法收集草酸钙肾结石患者肾脏组织标本21例,应用逆转录-聚合酶链反应(RT-PCR)对肾脏组织中的VKORCl进行扩增,经过PCR产物纯化,通过直接测序和克隆测序的方法分析得到肾脏组织中VKORCl的剪接异构体。结果应用RT—PCR的方法成功地在草酸钙尿石症患者肾脏组织中扩增出VKORCl,并且得到2个剪接异构体,3%琼脂糖凝胶电泳显示1条约500bp的电泳条带(V1)和另1条约400bp的电泳条带(V2),经过测序鉴定可见两者碱基相差110bp,其余序列完全一致。结论草酸钙尿石症患者肾脏组织中存在VKORCl的剪接异构体。
Objective To analyze vitamin K epoxide reductase complex subunit 1 (VKORC1) in the kidney tissues of patients with calcium oxalate urolithiasis. Methods Twenty-one specimens of kidney tissues were harvested from urolithic patients. VKORC1 in the kidney was amplified by the reverse tran- scription-polymerase chain reaction (RT-PCR) , the PCR products were purified, and the VKORC1 gene splicing isoforms were analyzed by direct sequencing and clone sequencing. Results VKORC1 was successfully amplified in the kidney tissues of patients with calcium oxalate urolithiasis and two splice isoforms were obtained. 3% agarose gel electrophoresis showed two bands: about 500 bp (V1), about 400 bp (V2). After sequencing, there were 110 bp differences in bases, and the remaining sequences were identical. Conclusion In the kidney tissues of patients with calcium oxalate urolithiasis fine VKORC1 splicing isoforms.