Study Of ENO2/NLRP3 Inflammasome Pathway In Intestinal Injury Induced By Severe Acute Pancreatitis And Intervention Of Qingyi Pellet

Objective:To observe the role of γ-enolase(Enolase-2,ENO2)and NLRP3 inflammasome in intestinal mucosal barrier dysfunction in severe acute pancreatitis(SAP)and the intervention of Qingyi Pellet.Treatment of intestinal mucosal barrier injury provides experimental evidence.Methods:30 adult male SPF SD rats with a body weight of 180 g to 220 g were randomly divided into 5 groups:normal control group(CON group),severe acute pancreatitis model group(SAP group),ENO2 inhibition(NAF)treated group(NAF group),dexamethasone treated group(DEX group),and Qingyi pellet treated group(QYKL group).Each group 6 rats.In the CON group,the pancreas was gently turned several times after laparotomy;the SAP model were established by retrograde biliopanceatic duct injection with 5%sodium taurocholate solution(50mg/kg body weight).In the NAF group and the DEX group,NAF solution(dose:6.7mg/kg,concentration:1.34mg/ml)and dexamethasone solution(dose:10mg/Kg,concentration:5mg/ml)were respetivelly administered by intraperitoneal injection at 2h and 12h after modeling.The QYKL group was administered with the Qingyi pellet aqueous solution at 2h and 12h after the model was created.Rats in all groups were sacrificed and harvested 24 hours after surgery.The Pathological sections of the ileum and pancreas tissue were stained with hematoxylin-eosin staining(HE)and the corresponding pathological scores were obtained.Blood was taken from the abdominal aorta.Serum was extracted and the content of IL-1β,IL-18,TNF-α,D-lactic acid,and diamine oxidase(DAO)was determined by enzyme-linked immunosorbent assay(ELISA).The content of serum amylase were detected by Automatic biochemical analyzer;Western-Blot method to detect the expression of ENO2,NLRP3,Casepase-1 and other protein levels in intestinal tissues;qRT-PCR method to detect the levels of ENO2,NLRP3 genes in intestinal tissues expression.Results:Compared with the CON group,the pancreatic tissue of the SAP group was swollen and congested,the lobular structure was lost,a large number of inflammatory cells were infiltrated,the intestinal villi were atrophied,necrotic,and the intestinal mucosal structure was incomplete.The scores were all increased(p<0.05),consistent with the pathological characteristics of SAP intestinal mucosal barrier damage;the levels of serum amylase,IL-1β,IL-18,TNF-α,D-lactic acid,and DAO in the SAP group rats all increased.(P<0.05);the expression levels of ENO2,NLRP3,Casepase1 and other proteins in intestinal tissues increased(p<0.05);the expression levels of ENO2 and NLRP3 in intestinal tissues also increased(p<0.05).Compared with the SAP group,the indicators of the QYKL treated group,the NAF and DEX treated groups were improved to varying degrees.Conclusion:(1)SAP rat model produced by retrograde pancreaticobiliary duct injection with 5%sodium taurocholate is stable.Serum amylase,D-lactic acid and DAO are increased in model group rats.Pathological changes in pancreas and intestine are obvious.SAP-related intestinal mucosal barrier The damage model was successfully established;(2)ENO2 protein expression was increased in SAP group,ENO2 inhibitor NAF could inhibit the expression of ENO2 and NLRP3 inflammasome and its downstream inflammatory factors(IL-1β,IL-18);(3)Qingyi Pellet may improve intestinal mucosal function by regulating ENO2/NLRP3 signal pathway,thereby reducing acute intestinal mucosal barrier damage and inflammatory response in SAP.