锌指蛋白A20对大鼠腹膜透析相关性急性腹膜炎症反应的影响  

Effects of zinc finger protein A20 on the inflammatory reaction in rats with peritoneal dialysis-related acute peritonitis

作　　者：邹循亮[1] 鄢巨振[1] 徐刚[1] 吴平[1] 

机构地区：[1]杭州师范大学附属医院肾内科,杭州310015

出　　处：《重庆医学》2013年第31期3788-3791,共4页Chongqing medicine

基　　金：浙江省自然科学基金资助项目(Y2090522)

摘　　要：目的探讨锌指蛋白A20对脂多糖(LPS)诱导的大鼠腹膜透析相关性急性腹膜炎模型腹膜组织CD40表达、IκBα磷酸化及腹腔局部炎症反应的影响。方法将24只雄性SD大鼠随机分为4组,每组6只。对照组:腹腔注入4.25%葡萄糖乳酸盐腹膜透析液(简称腹透液,90mL/kg);LPS组:LPS 1mg/kg腹腔注入4h后,再注入腹透液;转染A20组及空载体组:超声微泡转染pGEM-T easy-A20或pGEM-T easy质粒至大鼠腹膜组织3d后相继注入LPS及腹透液。注入腹透液4h后处死大鼠,留取腹水、壁层及脏层腹膜组织。常规腹膜组织Masson、HE染色和腹水白细胞计数;Western blot检测腹膜组织A20、p-IκBα、IκBα、CD40蛋白的表达;RT-PCR检测腹膜组织CD40mRNA的表达;ELISA法检测腹水中IL-6水平。结果 LPS可诱导腹膜组织表达A20。与LPS组及空载体组相比,转染A20组大鼠腹膜组织水肿明显减轻、炎细胞浸润及腹水白细胞计数均减少,腹膜组织pIκBα/IκBα比值、CD40mRNA及蛋白表达、腹水IL-6水平均明显降低(P<0.05);与对照组相比,转染A20组腹膜组织p-IκBα/IκBα比值和腹水IL-6水平差异无统计学意义(P>0.05),但CD40mRNA及蛋白表达明显升高(P<0.05)。结论大鼠腹膜组织过表达A20可抑制LPS诱导的大鼠急性腹膜炎症反应,并对LPS信号通路中相关功能蛋白起一定的调控作用。Objective To explore the effects of zinc finger protein A20（A20） on the expression of CD40 and the phosphorylation of IkBα as well as the local inflammatory reaction of abdominal cavity in Sprague Dawley（SD） rats with peritoneal dialysis-related a- cute peritonitis induced by Iipopolysaccharide（LPS）. Methods 24 male SD rats were equally randomized to four groups （n = 6, each）. Control group： in}ected with 4.25 % dextrose peritoneal dialysate（PDF） via abdominal cavity （ 90 mL/kg） ; LPS group ： ini ec ted with LPS（1 mg/kg） via abdominal cavity 4 hour later followed by PDF injection; transfection A20 plasmid group and empty plasmid group：after transfer pGEM-T easy-A20 or pGEM-T easy plasmid via intraperitoneally using an ultrasound-microbubble- mediated system for 3 days,then injected with LPS and PDF via abdominal cavity. The rats were killed 4 hours after PDF injection. Peritoneum tissue was stained using Masson and HE. Leucocytes count in abdominal dropsy was performed. The proteins expres- sion of A20, p-IκBα,IκBα,and CD40 in peritoneum tissue were analyzed by western blot;the expression of CD40 mRNA in peritone- um tissue were determined by RT-PCR;IL-6 level in abdominal dropsy was determined by ELISA. Results LPS could induce the protein expression of A20 in rats peritoneum tissue. Compared with LPS group and empty plasmid group, the degree of edema,in- flammatory cells infiltration,and the ratio of p-IκBα,IκBα,mRNA and protein expression of CD40 in rats peritoneum as well as leu- cocyte counts and IL-6 level of abdominal dropsy were also significantly decreased in transfection A20 plasmid group（P〈0.05）; meanwhile,compared with control group,that of the ratio of p IκBα,IκBα and IL-6 level in transfection A20 plasmid group were no significant difference（P〉0.05）, but the mRNA and protein expression of CD40 were significantly higher than that of control group （P〈0.05）. Conclusion Over expression of A20 in SD rats peritoneum tissue could down-regulat

关 键 词：腹膜炎 腹膜透析 锌指蛋白A20 IΚBΑ CD40 

分 类 号：R459.5[医药卫生—治疗学]