The Therapeutic Effect Of Astragaloside Ⅳ On Peritoneal Membrane Remodelling By Inhibiting Peritoneal Mesothelial Cells NLRP3 Inflammasome Activation

Background/Aim:Peritoneal dialysis is one of the alternative treatments for end-stage renal disease.However,with the existing medical technology,long-term peritoneal dialysis will inevitably lead to peritonitis and peritoneal fibrosis.The mechanism of peritoneal dialysis related peritonitis and peritoneal fibrosis has not been clear.NLRP3 inflammasome is a molecular bridge between metabolic disorder and inflammation.It can mediate inflammatory response by identifying endogenous danger signals.In this experiment,adenine feed was used to induce chronic kidney disease mice,and then standard dialysate(4.25%dextrose)was used to induce them peritoneal fibrosis.Thus,this research aims to study the role of NLRP3 inflammasome in this process and the therapeutic effect of Astragaloside Ⅳ,a traditional Chinese medicine extract.In peritoneal fibrosis rats induced by peritoneal dialysate,inflammation and fibrosis related proteins were up-regulated,and the degree of fibrosis decreased in NLRP3 inhibition group and Astragaloside Ⅳ intervention group.Methods:In vivo:The mice model of chronic kidney disease was established by feeding 0.2%adenine feed,and then the peritoneal fibrosis mouse model was established by daily intraperitoneal injections with standard dialysate(4.25%dextrose).Observed the changes of NLRP3 inflammatory in the process of peritoneal fibrosis and the effect of Astragaloside Ⅳ on peritoneal fibrosis.Male C57BL/6J mice were randomly divided into 6 groups with 10 mice in each group.Control:normal feeding.CKD group:feed with 0.2%adenine and change to normal feeding on the 29th day.PD group:feed with 0.2%adenine and change to normal feeding on the 29th day;From the 15th day,intraperitoneal injection peritoneal dialysis with 100ml/kg/day for 28 days.NLRP3 inhibition(inf-39)group:0.2%adenine diet,changed to normal feeding on the 29th day;From the 15th day,inject intraperitoneally peritoneal dialysis with 100ml/kg/day for 28 days,and inf-39(12.5mg/kg)was injected intraperitoneally every 7 days.Low dose Astragaloside Ⅳ group:0.2%adenine feed,changed to normal feeding on the 29th day;From the 15th day,inject intraperitoneally(100ml/kg/day+Astragaloside 2.5mg/kg/day)for 28 days.High dose Astragaloside Ⅳ group:0.2%adenine feed,changed to normal feeding on the 29th day;From the 15th day,inject intraperitoneally(100ml/kg/day+Astragaloside 10mg/kg/day)for 28 days.On the day 0,28 and 42,take about 250ul blood from the tail tip.On 42th day,isoflurane anesthesia was used,take 400-500ul blood from the eyeball,and the blood urea nitrogen of mice was measured to verify the chronic kidney disease model.Observe pathomorphological changes in mice peritoneum with Masson staining,and t Detect experession of caspase-1、IL-1β、NLRP3 and vimentin.Detecte E-cadherin、TGF、NLRP3,cleved-caspase-1,caspase-1、IL-1β,Changes in IL-18 by Western blot.Results:①Standard PD solution induced PF mice with chronic kidney disease.In PD group,Masson staining showed mesothelial layer denudation,mesothelial cells partially fell off,inflammation,fibrous capsule formation and collagen proliferation around the peritoneum.The expression of caspase-1,cleved-caspase-1,NLRP3,vimentin、TGF-β1、IL-1β and IL-18 were increased and the expression of E-cadherin was increased.Astragaloside Ⅳ treatment was beneficial to alleviate peritoneal lesions.②Standard PD solution induced activation of NLRP3 inflammatory and the expression of NLRP3,Cleved-caspase-1,caspase-1,IL-1β and IL-18 in PD group was increased.NLRP3 inhibitor INF-39 protects peritoneal tissue,reduces the expression of TGF-β and VIMENTIN.Astragaloside Ⅳ inhibited NLRP3 inflammatory activation and decreased the expression of NLRP3,IL-1β,IL-18,caspase-1,cleved-caspase-1.③Astragaloside Ⅳ inhibited the activation of NLRP3 inflammatory induced by Standard PD solution,The expression of caspase-1,cleved-caspase-1,NLRP3,VIMENTIN、TGF-β1、IL-1β and IL-18 were increased and the expression of E-cadherin was increased in Astragaloside Ⅳ group.Conclusion:1.NLRP3 inflammasome induces the occurrence of PD related PF.2.Astragaloside Ⅳ inhibited the activation of NLRP3 inflammatory in a dose-dependent manner,and then inhibited the process of PF related to PD.