CAJ | 학술논문

目的：研究高浓度尿素诱导人脑微血管内皮细胞系（ HBMECs ）产生炎性因子及其机制。方法以相同渗透压的甘露醇为对照，高浓度尿素（25 mmol／L）干预HBMECs 3、6、12和24 h后，免疫荧光法观察细胞内肿瘤坏死因子α（ TNF-α）和诱导型一氧化氮合酶（ iNOS）的表达。蛋白免疫印迹法（ Western blot ）检测TNF-α、iNOS、环氧合酶-2（cycloxygenase-2，COX-2）、核因子κB（NF-κB）／P65和p-P65的表达水平。一氧化氮（NO）试剂盒检测细胞NO含量。结果高浓度尿素增强细胞内TNF-α和iNOS表达。细胞TNF-α、COX-2和p-P65蛋白水平在3和6 h明显高于对照组（ P＜0.01）；iNOS蛋白水平持续增高（ P＜0.01）。 NO含量在3 h明显增多（ P＜0.05）。结论高浓度尿素诱导人脑微血管内皮细胞产生炎性因子。
목적：연구고농도뇨소유도인뇌미혈관내피세포계（ HBMECs ）산생염성인자급기궤제。방법이상동삼투압적감로순위대조，고농도뇨소（25 mmol／L）간예HBMECs 3、6、12화24 h후，면역형광법관찰세포내종류배사인자α（ TNF-α）화유도형일양화담합매（ iNOS）적표체。단백면역인적법（ Western blot ）검측TNF-α、iNOS、배양합매-2（cycloxygenase-2，COX-2）、핵인자κB（NF-κB）／P65화p-P65적표체수평。일양화담（NO）시제합검측세포NO함량。결과고농도뇨소증강세포내TNF-α화iNOS표체。세포TNF-α、COX-2화p-P65단백수평재3화6 h명현고우대조조（ P＜0.01）；iNOS단백수평지속증고（ P＜0.01）。 NO함량재3 h명현증다（ P＜0.05）。결론고농도뇨소유도인뇌미혈관내피세포산생염성인자。
Objective To explore high concentrations of urea-induced human brain microvascular endothelial cell line( HBMECs) to produce inflammatory cytokines and possible mechanism .Methods HBMECs were incubated in high concentrations of urea or mannitol ( as osmotic control ) for 3,6,12 and 24 hours.Expression of TNF-αand iNOS was observed by immunofluorescence .Western blot analysis was employed to assess the protein expressions of TNF-α, iNOS, COX-2, NF-κB/P65 and p-P65.NO concentration was determined by a commercial NO assay kit . Results Immunofluorescence showed high positive immunostaining of TNF-αand iNOS after incubation in high concentration of urea stimulued as compared with control group .The protein expressions of TNF-α, COX-2 and p-P65 were significantly increased at 3 and 6 hours after high urea treatment (P<0.01), and iNOS was continued to increase from 3 to 24 hours ( P<0.01 ) .Moreover , NO content was increased at 3 hours after high urea treatment ( P<0.05 ) .Conclusions High concentration of urea can induce HBMECs to produce inflammatory cytokines .