Effects Of Adenovirus Interfering With PBEF Expression Preconditioning On Sodium-water Transport System After Cardiopulmonary Bypass And Its Mechanism Research

Objective:Pre-B-cell clone enhancement factor(PBEF)is an enzyme that is widely expressed in cells,and its expression changes are related to acute lung injury(ALI).The hypoxia-reoxygenation model of alveolar epithelial cells and pulmonary vascular endothelial cells was established in an in vitro cell experiment,and it was proved that they increase the permeability of pulmonary vascular epithelial cells and inhibit the sodium-water transport system of alveolar epithelial cells,these two aspects have led to an increase in pulmonary edema.Sodium-water transport system as the central link in the removal of pulmonary edema fluid,the relationship between PBEF and it in rat cardiopulmonary bypass(CPB)model and the mechanism of its effect are not clear.Therefore,in this study,adenovirus was used to interfere with the expression of rat PBEF,on this basis,an cardiopulmonary bypassmodel was constructed,and the researchde the relationship between PBEF and sodium-water transport system after CPB and the mechanism of action.Methods:(1)SD rats were randomly divided into the following 6 groups and treated accordingly: Group A(blank group,no treatment after anesthesia,),Group B(surgery group,deep hypothermia circulatory assisted cardiopulmonary bypass for 30 min),group C(lung injury group,constructed model of lung injury by injected LPS in tail vein),group D(adenovirus interference+ lung injury group,adenovirus-PBEF-shRNA vector was injected 1 week in advance,the rest of the operation is the same as group C),group E(adenovirus negative control + surgery group,injection 1 week in advance Adenovirus-blank shRNA vector,the rest operation is the same as group B),group F(adenovirus interference + surgery group,adenovirus-PBEF-shRNA vector was injected 1 week in advance,the rest operation is the same as group B).(2)Measure the dry-wet-to-weight ratio of lung tissue,cell NKA enzyme hydrolysis activity,lung tissue pathological changes,immunohistochemical detection of PBEF expression and localized expression.(3)Lung tissue was digested for Western blot and PCR to detect the expression of PBEF and sodium water channel-related proteins(ENaC,AQP1,AQP5)and pulmonary surfactant(SP)in lung tissue.(4)Detection of phosphorylated P38 MAPK,ERK1 / 2,Akt expression changes.Results:(1)Compared with the traditional model of lung injury induced by tail vein injection of LPS,rats undergoing CPB also cause a certain degree of lung injury.PBEF is highly expressed in both lung injury models.(2)In CPB lung injury model,the expression of phosphorylated MAPK,ERK1 / 2 and Akt increased,and the expression of sodium-water transport system related proteins increased.(3)After CPB rat model interfered with PBEF expression,the expression of phosphorylated MAPK,ERK1 / 2 and Akt decreased;the protein and mRNA expression of ENaC increased,the protein expression of AQP1 decreased,the protein and mRNA expression of AQP5 decreased.(4)After the CPB rat model interfered with the expression of PBEF,the NKA enzymatic hydrolysis activity increased,and the dry-wet-to-weight ratio of lung tissue decreased.Conclusion:(1)PBEF is a marker protein for lung injury after cardiopulmonary bypass.(2)PBEF can regulate the expression of sodium water transport system related factors through phosphorylation of MAPK,ERK1 / 2 and Akt.(3)Inhibition of PBEF expression can reduce pulmonary edema after cardiopulmonary bypass through improve activity of ENaC and NKA.