吡咯烷二硫代甲酸铵干预NF—κB p65核转位抑制尿毒症大鼠主动脉钙化  

Pyrrolidine - dithio - carbamate ammonium counteracts high - phosphate - induced aortic calcification by inhibiting p65 nuclear translocation in uremic rats

作　　者：于亚民 贾俊亚[1] 闫铁昆[1] 高姗[1] 商文雅[1] 韦丽[1] 李红芬[1] 林珊[1] 

机构地区：[1]天津医科大学总医院肾内科,300052

出　　处：《中华肾脏病杂志》2014年第8期614-618,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金（31270791,30800529）;天津市科委应用基础及前沿计划（10JCYBJC11700）;天津市卫生局重点攻关课题（11KGl32）

摘　　要：目的探讨NF．KB抑制剂吡咯烷二硫代甲酸铵（PDTC）对尿毒症大鼠主动脉钙化的干预作用及相关机制。方法16只雄性sD大鼠被随机分为尿毒症模型组及PDTC干预组，均予0．75％腺嘌呤及高磷（1％）饮食，制备尿毒症动脉钙化模型，干预组同时腹腔注射PDTC100mg·kg^-1·d^-1。另取8只匹配大鼠作为健康对照。8周后处死大鼠，HE及yon Kossa染色观察腹主动脉病理改变及钙化情况，免疫组织化学方法检测骨调素（OPN）、核心结合因子α1（Cbfα1）在主动脉的定位与表达，Western印迹检测主动脉NF-κB总p65与细胞核内P—p65、OPN、Cbfα1等蛋白表达量。结果造模4周及8周，尿毒症组、PDTC组大鼠血清BUN、Scr、血磷、钙磷乘积均显著高于对照组（均P＜0．01），但此两组间差异无统计学意义（P，0．05）；造模8周，尿毒症组和PDTC组大鼠主动脉明显增厚及中膜钙化，PDTC组程度较轻（P＜0．05），免疫组化显示主动脉内皮下、中膜及外膜均有OPN、Cbfα1表达，PDTC组表达量较少（均P＜0．05）；Western印迹显示PDTC组主动脉NF—κB总p65、核p-p65、OPN、Cbfα1表达均较尿毒症组下降（均P＜0.01），且Cbfcd表达与p65、核P—p65表达呈正相关（r=O．707，P＜0．01；r=O．507，P＜0．01）。结论PDTC可阻断NF-κBp65核转位，抑制尿毒症大鼠主动脉Cbfcd表达，减轻血管钙化。Objective To investigate the effects of pyrrolidine-dithio-carbamate ammonium （PDTC） on high-phosphate-induced vascular calcification in uremic rats. Methods Eight-week-old SD rats were pair-fed with standard chow containing 1.2% calcium and 0.6% phosphorus for the control group （n=8） or 0.75% adenine, 1.2% calcium, and 1% phosphorus for the chronic renal failure（CRF） group （n=8） or PDTC group （intraperitoneal injection, 100 mg·kg^-1·d^-1, n=8） for 8 weeks. The abdominal aortas were excised for Western blotting and immunostaining assay of NF-κB p65, osteopontin （OPN） and core binding factor αl（Cbfctl） protein. Results Serum urea nitrogen, creatinine, inorganic phosphate, calcium-phosphorus product increased significantly in CRF group and PDTC group after 4 weeks and 8 weeks （all P 〈 0.01）, although no differences were found between the latter two groups. After 8 weeks, aortic calcification was found in these two groups, immunostaining assay revealed OPN and Cbfcd expressed in aortic intima, media and adventitia, and Western blottinganalysis showed that total NF- κB p65, nuclear phosphorylated- p65 （p- p65）, OPN and Cbfcil expressions were significantly higher than those in control group （all P 〈0.01）. The expression of total p65 and p- p65 was positively correlated with Cbfo^l（r=0.707, P 〈 0.01; r=0.507, P 〈0.01）. Conclusion PDTC alleviates inorganic phosphate- induced aortic calcification significantly by inhibiting the nuclear translocation of NF-κB p65 and the expression of Cbfal.

关 键 词：尿毒症 NF-κB 血管钙化 吡咯烷二硫代甲酸铵 

分 类 号：R692.5[医药卫生—泌尿科学]