Expression Of NALP3 Inflammatory And Dahuangfuzi Decoction Experimental Study In Lung Injury Of Severe Acute Pancreatitis Rats

Objective: The aim of this study was to investigate whether Dahuangfuzi decoction,a prescription in traditional Chinese medicine,could relieve lung injury by regulating NALP3 in the signal pathway after severe acute pancreatitis induced by sodium taurocholate in rats,and to elucidate the role AQP-5,IL-1? and NALP3 play in the pathogenesis of severe acute pancreatitis lung injury.Methods: A total of 24 SPF healthy male SD rats were randomly divided into 4 groups:blank control group(n=6),SAP Group(n=6),Dahuangfuzi decoction treatment group A(n=6),Dahuangfuzi decoction treatment group B(n=6).SAP models were established by retrograde injections of 4% sodium taurocholate(1 ml / kg)solutions into rat pancreaticobiliary ducts;Treatment group A were enemaed by Dahuangfuzi decoction 12 hours after surgery.Treatment group B were enemaed 12,24 h after surgery.Blood samples were taken from the abdominal aorta for measuring endotoxin,serum amylase and serum calcium concentrations and afer taking blood samples the rats were sacrificed.The morphological changes of lung tissue and pancreas were observed under microscope.The pathological evaluation of lung tissue was scored by Smith scoring method.The expression of IL-1? in serum and lung tissue were detected by ELISA.The expression of AQP-5 and NALP3 protein in lung tissue were detected by immunohistochemistry,and Western blot was also added to detect the expression and distribution of NALP3 protein in lung tissue.Results: 1.The serum endotoxin levels in rats:Compared with the blank control group,the serum endotoxin levels in SAP group were significantly increased(P <0.05).Compared with the SAP group,the plasma endotoxin level in the treatment group was lower(P <0.05),and serum endotoxin in Treatment group B was lower than that in group A(P<0.05).2.Rat serum amylase,serum calcium content:Compared with the blank control group,the amylase content in the SAP group was significantly higher(P <0.05).Contrast to SAP group,the content of serum amylase decreased while the serum calcium content increased(P<0.05).The levels of amylase in group B were lower than that in group A and the difference was statistically significant(P <0.05).3.Observation of Lung and Pancreas histologic changes under HE Staining:Sham group exerted normal structures without histopathologic changes.In comparison with the sham group,lung tissues from the model group showed a severe pathologic abnormality,including pulmonary interstitial hyperemia,edema and hemorrhage,infiltration of inflammatory cells into alveolar space,and alveolar collapse;However,those pathologic changes were significantly ameliorated after the enema of Dahuangfuzi decoction.In treatment group A,gland edema was lighten,Inflammatory cell infiltration,local bleeding and necrosis was decreased;In treatment group B,the pancreas only had mild edema,slightly widened tissue gap and a small amount of inflammatory cell infiltration.The four groups had significant differences in injury scores(P <0.05).4.Results of ELLSA: Compared with the blank control group,the levels of IL-1? in serum and lung tissue of SAP group had a higher expression(P <0.05).Compared with SAP group,the level of IL-1? in treatment group was lower than that in model group(P <0.05).Treatment group B compared with treatment group A had a decreased expression(P<0.05).5.Immunohistochemical results: Compared with the blank control group,the expression of NALP3 in the SAP group was higher(P<0.05).The expression of NALP3 was decreased while AQP5 was upregulated in the treatment group contrast by SAP group(P <0.05).6.Western-blot results:The expression of NALP3 in the SAP group was higher than the blank control group(P<0.05).The expression of NALP3 in the treatment group was lower than that in the SAP group(P<0.05).Conclusion:(1)The expression of NALP3 increased during the formation of acute lung injury in severe acute pancreatitis,and the expression of IL-1? was increased,leading to the decrease of AQP-5 expression,resulting in water transport disorder and increased water content in the lung.Therefore,NALP3 inflammatory body and AQP-5 may be involved inthe pathogenesis of acute lung injury in severe acute pancreatitis.(2)Rhubarb Dahuangfuzi decoction may reduce the expression of IL-1? and decrease the expression of AQP-5 by reducing endotoxin and down-regulating NALP3 inflammatory body,thus improving the degree of lung injury in severe acute pancreatitis.