CAJ | 학술논문

目的：探讨锌指蛋白A20对脂多糖（L PS ）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，90 mL/kg）；LPS组：LPS 1 mg/kg腹腔注入4 h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3 d后相继注入LPS及腹透液。注入腹透液4 h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CD40 mRNA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p-IκBα/IκBα比值、CD40 mRNA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CD40 mRNA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L PS诱导的大鼠急性腹膜炎症反应，并对L PS信号通路中相关功能蛋白起一定的调控作用。
목적：탐토자지단백A20대지다당（L PS ）유도적대서복막투석상관성급성복막염모형복막조직CD40표체、IκBα린산화급복강국부염증반응적영향。방법장24지웅성SD대서수궤분위4조，매조6지。대조조：복강주입4．25％포도당유산염복막투석액（간칭복투액，90 mL/kg）；LPS조：LPS 1 mg/kg복강주입4 h후，재주입복투액；전염A20조급공재체조：초성미포전염pGEM-T easy-A20혹pGEM-T easy질립지대서복막조직3 d후상계주입LPS급복투액。주입복투액4 h후처사대서，류취복수、벽층급장층복막조직。상규복막조직Masson、HE염색화복수백세포계수；Western blot검측복막조직 A20、p-IκBα、IκBα、CD40단백적표체；RT-PCR검측복막조직CD40 mRNA적표체；ELISA법검측복수중IL-6수평。결과 LPS가유도복막조직표체A20。여LPS조급공재체조상비，전염A20조대서복막조직수종명현감경、염세포침윤급복수백세포계수균감소，복막조직p-IκBα/IκBα비치、CD40 mRNA급단백표체、복수IL-6수평균명현강저（P＜0．05）；여대조조상비，전염A20조복막조직p-IκBα/IκBα비치화복수IL-6수평차이무통계학의의（P＞0．05），단CD40 mRNA급단백표체명현승고（P＜0．05）。결론대서복막조직과표체A20가억제L PS유도적대서급성복막염증반응，병대L PS신호통로중상관공능단백기일정적조공작용。
Objective To explore the effects of zinc finger protein A20(A20) on the expression of CD40 and the phosphorylation of IκBαas well as the local inflammatory reaction of abdominal cavity in Sprague Dawley (SD) rats with peritoneal dialysis-related a-cute peritonitis induced by lipopolysaccharide (LPS ) .Methods 24 male SD rats were equally randomized to four groups (n= 6 , each) .Control group:injected with 4 .25% dextrose peritoneal dialysate(PDF) via abdominal cavity (90 mL/kg);LPS group :injec-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/IκBαand 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-regulate the inflammatory reaction in peritonitis induced by LPS ,which might be involved in modulating the expression of associated functional protein during LPS signal pathw ay .