Protective Effects Of Albumin On Vascular Endothelial Cell In Severe Acute Pancreatitis

Objective:On the basis of observing the physiopathology changes of severe acute pancreatitis in rats, to investigate the relationship of vascular endothelial injury and organ damage under the effect of trypsin in vitro and vivo experiment,and confirm the protective effect of albumin on vascular endothelial injury initially.Methods:1. Nienty rats were randomly divided into sham operation and model groups,and each group were randomly divided into3h、6h、12h groups. The severe acute pancreatitis model was constructed by retrograde pancreatic duct infusion of Na-taurocholate.The sham operation group only turn over duodenum and pancreas,don’t inject Na-taurocholate.Observe the state and survival rate of each group after operation, corresponding group was executed at the time of3h、6h、12h,the expression of AMY、AST、ALT、BUN、Cre and ALB in the serum were detected,pathological changes of pancreas、lungs、liver、kidney、heart、intestine and thoracic aorta tissues were observed. vascular endothelial cells were stained with a PE Mouse Anti-Rat CD31monoclonal antibody for flow cytometric analysis.2. Rat VECs were isolated and cultured,put the3rd-5th generation cells into96-well microplate, Change serum free medium200μl when the VECs overspread the bottom of microplate, ten different concentrations of trypsin50μl added in the microplate (dilute the concentration of trypsin from0.25%to0.00048828125%using the doubling dilution method),each concentration use8wells and were incubated for1hour, The cell proliferation ability was measured by MTT.3. Put the3rd-5th generation cells into96-well microplate, Change serum free medium100μl when the VECs overspread the bottom of microplate, ten different concentrations of albumin50μl added in the microplate(dilute the concentration of albumin from10%to0.02%using the doubling dilution method),each concentration use8wells, each well added in0.125%trypsin50μl and were incubated for1hour, the cell proliferation ability was measured by MTT.Results: 1. The survival rate of SO group is100%and the SAP group is78%,the pathological score and the expression of serum AMY、AST、ALT、BUN and Cre in the SAP group is significantly higher than the SO group at the same time point, the SAP group’s expression of serum ALB in12h is significantly lower than the SO group. vascular endothelial injury was observed by pathobiology in the SAP rat’s organs. With the increasing time,all those changes of biochemical index and pathobiology increased significantly.2. The VEC in the blood of the SAP group increase significantly in number of quantity than the SO group,the quantity of VEC has a positive correlation with AMY in the serum and pancreas pathological score.3. With the increasing concentration of trypsin, OD value decreased gradually, The OD values have a negative correlation with the concentrations of trypsin; With the increasing concentration of albumin, OD value increased gradually, The OD values have a positive correlation with the concentrations of albumin.Conclusions:1. The vascular endothelial injury damaged by trypsin is the pathological basis of organ damage in acute severe pancreatitis.2. Albumin can protect the vascular endothelial injury damaged by trypsin, and the protective effect of albumin on VEC has a positive correlation with albumin concentration.