清解化攻方对急性胰腺炎小鼠腺泡细胞焦亡和肠屏障的影响OA北大核心CSTPCD

Objective To explore the mechanism of regulating circRNA homeodomain-interacting protein kinase 3(circHipk3)by Qingjie Huagong Decoction(QJHGD)to alleviate acute pancreatitis(AP)alveolar cell scorching and protect the intestinal barrier in acute pancreatitis.Methods Sixty mice were randomly divided into the blank group,model group,ustatin group,QJHGD group,gene silencing group,and gene silencing combined with Chinese medicine(CM)group,10 in each group.Gene silencing,gene silencing combined with CM group received injections of circHipk3 gene silencing adeno-associated virus through the tail vein.Model and each intervention group received injections of 50 µ g/kg caerulein once every hour for seven days.The blank and model group received an equal amount of distilled water,the gene silencing combined with CM group received 2 g/kg of CM by gavage,and the ustekin group received an intraperitoneal injection of ustatin 5 U/kg,both twice daily for three days.The changes of pancreas tissue and ileal pathology were observed.The levels of serum endotoxin,tumor necrosis factor α(TNF-α),interleukin 18(IL-18),and IL-1β were detected.The expression of pancreatic siderophore D(GSDMD),ileal occluding small band protein 1(ZO-1)and Occludin were detected.The express of pancreatic cysteine protease-1(caspase-1),GSDMD,NOD-like receptor heat protein domain-related protein 3(NLRP3),IL-1β,ZO-1,Occludin,closure protein 1(CLDN1)protein were detected using Western Blot.The levels of pancreatic circHipk3,microRNA miR-193a-5p,caspase-1,and GSDMD genes were detected by quantitative real-time Polymerase Chain Reaction(qPCR).Results Compared with the control group,the ascites,wet/dry proportion(W/D),pancreatic and ileal tissue scores,serum endotoxin,TNF-α,IL-18,and IL-1 levels,and GSDMD positive expression increased,the expression of GSDMD,caspase-1,NLRP3,and IL-1 protein increased,ZO-1,Occludin,and CLDN1 expression decreased.Meanwhile,the expression of CircHipk3,caspase-1,and GSDMD mRNA increased,miR-193a-5p,ZO-1,and Occludin expression decreased,intestinal permeability increased in the model group(all P＜0.05).Compared with the model group,the W/D of pancreas decreased in the intervention groups.The pancreas ascites,TNF-a,IL-18,IL-1,endotoxin levels,and positive expression of GSDMD decreased.Intestinal permeability increased,circHipk3,caspase-1,and GSDMD mRNA expression decreased,ileal protein expression increased,GSDMD,Caspase-1,and IL-1 protein expression increased,miR-193a-5p expression increased,and ZO-1 and Occludin positive expression increased in the ustatin group,QJHGD group,and gene silencing combined with CM group(all P＜0.05).Compared with the gene silencing group,the W/D and pancreas ascites,scores of pancreas and ileum tissue decreased,and protein expression of ZO-1,occludin,and CLDN1 increased.Additionally,the expression ofcircHipk3,caspase-1,GSDMD mRNA,miR-193a-5p,ZO-1,and occludin increased,as well as intestinal permeability decreased in the gene silencing combined with CM group(all P＜0.05).Conclusion QJHGD could down-regulate the expression levels of circHipk3 and caspase-1 genes,up-regulate miR-193a-5p,reduce the expression of Pyroptosis protein,and reduce pancreatic damage to protect the intestinal barrier.