肿瘤抑制蛋白CYLD在急性坏死性胰腺炎大鼠肺损伤中的作用  

The role of tumor suppressor factor CYLD in lung injury associated with acute necrotizing pancreatitis

作　　者：靳杯[1] 刘立民[1] 闫文貌[1] 程石[1] 

机构地区：[1]首都医科大学附属北京天坛医院普通外科,100050

出　　处：《中华肝胆外科杂志》2013年第1期58-61,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：北京市卫生系统高层次卫生技术人才队伍建设专项培养基金（2011-3-033）

摘　　要：目的 研究肿瘤抑制蛋白CYLD在急性坏死性胰腺炎（ANP）肺损伤中的表达状况及其对核转录因子-κB（NF-κB）的影响,探讨其在肺损伤中的作用.方法 72只成年SD大鼠随机分成假手术组、ANP组及氯化钆（GdC13）治疗组,每组24只.采用逆行胰胆管注射5％牛磺酸钠的方法建立ANP大鼠模型.各组术后1、3、6、12 h分批处死动物.经支气管肺泡灌洗获取肺泡巨噬细胞（AM）,检测支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）和白细胞介素1β（IL-1β）水平,同时行肺组织病理学检查.蛋白免疫印迹法测定各组AM中CYLD及NF-κB活性水平.结果 ANP大鼠肺损伤随着病情进展而逐渐加重.AM分泌TNF-α、IL-1β水平逐渐升高,至6h达到高峰,分别为（491.3±20.3）ng/L和（178.83±11.32） ng/L,12 h又回落.ANP组TNF-α及IL-1β含量较假手术组明显升高（P＜0.05）,而GdCl3治疗组较ANP组明显降低（P＜0.05）.NF-κB蛋白在假手术组表达呈低水平,而CYLD蛋白表达呈高水平.ANP组NF-κB蛋白表达从3h开始与假手术组相比已有上升,差异有统计学意义（P＜0.05）,并随时间进行,表达逐渐增高.ANP组CYLD蛋白表达则相反,与假手术组相比从3h开始已有下降（P＜0.05）,随时间进行,表达逐渐减少.GdCl3治疗组CYLD及NF-κB蛋白表达变化趋势与ANP组趋势相近,CYLD蛋白表达与ANP组相比,从6h起表达量开始增加（P＜0.05）；NF-κB蛋白表达则从1h起表达量明显下调,与ANP组相比差异有统计学意义（P＜0.05）.ANP组、GdCl3治疗组AM中NF-κB与CYLD的表达活性均呈负相关（r分别为-0.918,-0.723,P＜0.01）.结论 CYLD在ANP肺损伤中呈低度表达,且具有明显时相,从3h起表达逐渐减低；而NF-κB则相反,随时间进展,表达逐渐增高.经GdCl3处理后,CYLD表达增强,NF-κB表达降低,表明在ANP时CYLD的表达对NF-κB表达也有负性调控作用,进而使肺损伤减轻.Objective Acute necrotizing pancreatitis （ANP） may cause lung injury.This study explores two factors that are associated with lung damage from ANP,the expression of tumor suppressor factor CYLD and nuclear factor-kappa B （NF-κB）.Methods 72 adult Sprague-Dawley rats were randomly divided into 3 groups：sham operation,ANP,and GdCl3 treatment groups （n=24 for each group）.A retrograde injection of 5% sodium taurocholate into the biliopancreeatic duct of rats induced ANP,and the animals were killed 1,3,6,and 12 hours after the ANP induction.AMs were harvested by bronchoalveolar lavage technique,and TNF-a and IL-1β levels in bronchoalveolar lavage fluid （BALF） were evaluated.Lung tissue was checked with histological examinations,and the activity of NF-κB and CYLD in AM were measured by western blot.Results TNF-α and IL-1β secreted by AM were gradually elevated,peaked on the sixth hour,had maximums of （491.3 ±20.3）ng/L and （178.83±11.32）ng/L respectively,and decreased on the twelfth hour.The levels of TNF-α and IL-1β in the ANP group were significantly higher than the sham operation group （P〈0.05）,and the GdC13 group levels were obviously lower than ANP group.In the sham operation group,the expression of NF-κB was low and CYLD was high.In the ANP group,when compared to the sham operation group,the expression of NF-κB rose after 3 hours and continued to rise with time progression （P〈0.05）.In contrast,CYLD protein expression in the ANP group dropped after 3 hours and continued to gradually decrease （P〈0.05）.The CYLD and NF-κB protein expression in GdCl3 groups had similar trends as the ANP group.GdCl3 group CYLD levels began to rise at 6 hours （P〈0.05）,and NF κB levels began to fall at 1 hour （P〈0.05）.The expression of NF-κB and CYLD possessed a negative correlation in both the ANP and GdCl3 groups （r =-0.918,r=-0.723,P〈 0.01）.Conclusions Therefore,in acute lung injury associated with acute pancreatitis,CYLD expression decreased with evident phases

关 键 词：胰腺炎 急性坏死性 肿瘤抑制蛋白质类 肺 创伤和损伤 

分 类 号：R657.51[医药卫生—外科学]