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

ABSTRACT

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 groupinjected with 4 .25% dextrose peritoneal dialysate(PDF) via abdominal cavity (90 mL/kg);LPS groupinjec-ted with LPS(1 mg/kg) via abdominal cavity 4 hour later followed by PDF injection ;transfection A20 plasmid group and empty plasmid groupafter 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(P0 .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 .