The Expression And Relationship Of MLCK And Adherin Junction In Early Hypertriglyceridemia Associated Acute Pancreatitis

Objective To establish a rat model of acute necrotizing pancreatitis by retrograde injection of 4%sodium taurocholate into the duodenum pancreatic duct and observe the effects of high triglyceride on inflammatory markers,pancreatic pathological injury,and intercellular adhesion in acute necrotizing pancreatitis rats.Effects of adherin junction,AJ),and protein expression of myosin light chain kinase（MLCK）and their relationship.Methods A total of 144 male SD rats were fed adaptively for 3 days and randomly divided into normal diet group（group A,n=48）and high-fat diet group（group B,n=96）.After 4 weeks of feeding,the A component was divided into 2 subgroups:normal group（N group,n=24）and acute necrotizing pancreatitis group（ANP group,n=24）;B component was 4 subgroups.Respectively,hypertriglyceridemiagroup（HTGgroup,n=24）,hypertriglyceridemicpancreatitisgroup（HTGPgroup=24）,hypertriglyceridemia ML-7 intervention group（HTG+ML-7 group,n=24）,hypertriglyceridemic pancreatitis ML-7 intervention group（HTGP+ML-7group,n=24）.N group,HTG group and HTG+ML-7 group were sham-operated control group,ANP group,HTGP group,HTGP+ML-7 group were model group,and 4%of bovine gallbladder was injected through retrograde duodenal papilla.A model of acute necrotizing pancreatitis was established by sodium citrate.Rats were sacrificed at 3,6,and 12 hours after modeling.The rats were observed for pancreatic and intraperitoneal conditions.Serum amylase and serum calcium levels were measured by automatic biochemistry analyzer;HE staining was used.The changes of pancreas morphology were observed;the expression of MLCK protein in pancreas was detected by Western blot;the localization and expression of E-cadherin,β-catenin and MLCK proteins were detected by immunohistochemistry;The level of serum IL-10,IL-6 were detected by ELISA.Results 1.After feeding for 4 weeks,the body weight of normal rats was200.02±5.07g,and that of rats fed high-fat diet was 235.43±28.39g.The difference was statistically significant（P<0.05）.2.Ordinary feed group and high fat The serum TG levels in the normal group were（0.41±0.10）mmol/L and（2.11±0.53）mmol/L,respectively;the difference was statistically significant（P<0.05）;the serum TC content was（2.60±0.30）mmol/L,respectively.（2.61±0.45）mmol/L,the difference was not statistically significant（P>0.05）;3.Histopathological scores of the pancreas gradually increased as the course progressed,and there was a statistically significant difference between the point model group and the control group（P<0.05）,HTGP group,HTGP+ML-7group pancreatic pathological score was significantly higher than ANP group（P<0.05）;4.Simultaneous model group compared with the control group serum amylase,the difference was statistically significant（P<0.05）,serum Ca²⁺began to decline 6 hours after modeling,compared with the control group was statistically significant（P<0.05）,while at the same time between the model group serum AMY,serum Ca²⁺was not statistically significant（P>0.05）;5.The expression of MLCK in the pancreatic tissue of the ANP group,the HTGP group and the HTGP+ML-7 group was significantly up-regulated by Western blot compared with the N group,the HTG group,and the HTG+ML-7 group.P<0.05),ML-7 can inhibit the expression of MLCK;6.immunohistochemistry detection of rat pancreas,HTGP group compared with ANP group,E-cadherin,β-catenin,MLCK protein content increased significantly（P<0.05）;E-cadherin,β-catenin,and MLCK were positively correlated with pathological scores of the pancreas:r1=0.644,r2=0.474,r3=0.685（P<0.05）;HTGP+ML-7 The degree of pancreatic injury in group 7 was lower than that of HTGP group（P<0.05）,the expression of E-cadherin,β-catenin,and MLCK protein was decreased（P<0.05）;serum IL-6 and IL-10 began to increase at 3 hours after model establishment.There was a significant difference between each model group and the control group（P<0.05）.With the prolongation of modeling time,the concentrations of IL-6 and IL-10 gradually increased.Serum IL-6 and IL-10 levels in the HTGP group were significantly lower than those in the SAP group at the same time point（P<0.05）.Serum IL-6 and IL-10 levels in the HTGP+ML-7 group were lower than those in the HTGP+ML-7 group at the same time point.In the group,the difference was significant（P<0.05）.Conclusions 1.Rat model of hypertriglyceridemic acute pancreatitis with hypertriglyceridemia can be successfully induced by high-fat diet fed with retrograde duodenal choline and pancreatic duct injection of 4%sodium taurocholate.2.Hypertriglyceridemia can aggravate early acute necrotizing pancreatitis in rats.3.MLCK may participate in pancreatic injury by regulating cell-cell adhesion in hypertriglyceridemia pancreatitis.