摘要
目的观察骨桥蛋白（OPN）在草酸钙肾结石形成中的作用。方法构建携带OPN-短发卡RNA（shRNA）并能有效沉默OPN基因的慢病毒载体，通过肾静脉导入大鼠肾脏，7d后开始喂饲含0．8％乙二醇和1％氯化铵的饮水，诱导肾脏草酸钙晶体沉积。喂饲9d后处死大鼠取出肾脏，比较体内沉默肾脏OPN基因表达后草酸钙晶体在肾组织中沉积数量的变化。结果携带OPN-shRNA的慢病毒载体均能有效感染肾脏皮质和髓质，OPN-shRNA能显著抑制肾组织OPN基因的表达。OPN-shRNA组与空病毒载体组。肾皮质草酸钙晶体（以每个观察视野结晶的平均总像素表示）分别为207164-7755和54921±3469（P〈0．01），髓质分别为18618±2285和30475±1735（P〈0．01）。结论沉默高草酸尿大鼠肾组织OPN基因的表达可以显著减少草酸钙晶体在肾脏中的沉积，提示OPN在草酸钙肾结石形成过程中可能起一定的促进作用。
Objective To address the effects of osteopontin （OPN） on calcium oxalate renal cal- culus formation. Methods We silenced the OPN gene in rat kidney in vivo using lentivirus vector - deliv- erd short hairpin RNA （shRNA） targeting OPN through renal venous transduction. On the day 7 after in- jection with lentivirus, rats were fed on water containing 0. 8% glycol and 1% ammonium chloride for 9 days. At the end of the experiments, the rats were killed and the kidney tissues were removed for subse- quent analysis. Results The lentivirus vector could effectively infect renal cortex and medulla tissue. OPN - shRNA transfection resulted in significant reduction in the content of OPN. Crystals recorded as the average total pixels （object area） were 20 716 ±7 755 and 54 921 ± 3 469 in renal cortex sections in the OPN -shRNA group and negative lentivirus vector group, respectively （P 〈 0. 01 ）, as well as 18 618 ± 2 285 and 30 475±1 735 in the medulla sections in both groups, respectively （P 〈0. 01 ）. Conclusion Specific knockdown of OPN gene in kidneys of hyperoxaluric rats leads to a decrease in OPN production and significantly inhabits calcium oxalate crystal deposition. OPN may play a certain role in promoting calcium oxalate renal calculus formation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第12期2675-2677,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81170651）
关键词
骨桥蛋白
草酸钙
肾结石
RNA干扰
Osteopantin
Calcium oxalate
Nephrolithiasis
RNA interference
