Trimetazidine For Acute Pancreatitis In Rats Of HIF-1 Alpha-iNOS-NO Signaling Pathways And HIF-1 Alpha VEGF Signaling Pathways

Objective: The study was aimd to study the protective effect of trimetazidine in rats of acute pancreatitis of HIF-1alpha-iNOS-NO signaling pathways and HIF-1 alpha VEGF signaling pathways.Methods: A totally 54 healthy and clean male SD rats(weight 240g-260g),adaptive feed after 1 week were randomly divided into three groups:the control group(group A,n = 18);the acute pancreatitis group(group B,n =18);the trimetazidine intervention group(group C,n = 18).The control group rats after laparotomy only turn the pancreas and duodenum gently;the acute pancreatitis group was made by retrograde injection of 4% sodium taurocholic acid into the biliary-pancreatic(1 mL/kg,0.2 mL/min);the trimetazidine treatment group was given by gavage with trimetazidine(5 mg/kg,dissolved in normal saline)after the rats awaken after making modes.Each group was divided into three subgroups at 6h,12 h and 24 h after making modes,and the rats were sacrificed at the above time points.Six rats were taken from each time point.The level of IL-6 in serum was measured by enzyme-linked immunosorbent assay(ELISA).The level of serum amylase and NO in serum was determined by colorimetry.The contents of VEGF and iNOS in pancreatic tissue were detected by immunohistochemistry.Western blot was used to detect the content of HIF-1α in pancreatic tissue.Pancreatic tissue and 10%formaldehyde were fixed to make paraffin sections for HE staining.The pathological changes of pancreas were observed under light microscope.Results:1 The level of serum amylase:The level of serum amylase in the control group is not obvious elevated;the level of serum amylase in the acute pancreatitis group increased since making modes;the level of serum amylasein the trimetazidine intervention group in 6 h,12 h,24 h showed a trend of gradual decline.Compared with the control group,the level of serum amylase in the acute pancreatitis group at each time point were significantly increased(P<0.05).Compared with model group,the level of serum amylase in the trimetazidine intervention group at each time point was significantly decreased(P<0.05).2 The level of IL-6: There was no significant difference in the level of IL-6 in the control group at each time point.The level of IL-6 in the acute pancreatitis group was significantly higher than that in the control group at each time point(P<0.05),and the level of IL-6 in the trimetazidine intervention group at each time point was significantly lower than the acute pancreatitis group(P <0.05).The level of serum IL-6 in the acute pancreatitis group was gradually increased at each time point(P<0.05).The level of IL-6 in the trimetazidine intervention group was gradually decreased at each time point(P<0.05).3 The level of serum NO: There was no significant difference in the level of serum NO in the control group at each time point.The level of serum NO in the acute pancreatitis group was significantly higher than that in the control group at each time point(P<0.05),and the level of serum NO in the trimetazidine intervention group at each time point was significantly lower than the acute pancreatitis group(P<0.05).The level of serum serum NO in the acute pancreatitis group was gradually increased at each time point(P<0.05).The level of serum NO in the trimetazidine intervention group was gradually decreased at each time point(P<0.05).4 The expression level of HIF-1 alpha: There was no significant difference in the expression level of HIF-1 alpha in the control group at each time point.The expression level of HIF-1 alpha in the acute pancreatitis group was significantly higher than that in the control group at each time point(P<0.05),and the expression level of HIF-1 alpha in the trimetazidine intervention group at each time point was significantly lower than the acute pancreatitis group(P <0.05).The level of expression level of HIF-1 alpha inthe acute pancreatitis group was gradually increased at each time point(P<0.05).The expression level of HIF-1 alpha in the trimetazidine intervention group was gradually decreased at each time point(P <0.05).5 Specimen of pancreatic tissues: The control group: There was no obvious edema,necrosis or bleeding performance,no fluid in the abdominal cavity.There was no obvious difference in the structure of the pancreatic lobules under light microscopy.The structure of the acini was intact,the interstitial had no edema,and a few inflammatory cells were infiltrated,and there was no significant difference at each time point.The acute pancratitis group: There was visible pancreatic tissue edema,a small amount necrosis of pancreatic gland follicular,a small amount of bloody effusion which can be seen in abdominal cavity.Pancreatic necrosis area with the extension of the modeling time gradually expanded,bleeding,calcification and bloody peritoneal fluid volume increased with time.There was a small amount of inflammatory cell infiltration,a small amount of interstitial hemorrhage,partial pancreatic acinar cell necrosis in 6h group of light microscopic;there was inflammatory cell infiltration,interstitial hemorrhage,acinar cell necrosis increasing in 12 h group of light microscopic;there was a large number of inflammatory cell infiltration,increased interstitial hemorrhage,pancreatic acinar cell necrosis,pancreatic lobular structure destruction,large pancreas parenchymal cell necrosis in 24 h group of light microscopic.The trimetazidine intervention group: There was pancreatic tissue congestion and edema,no obvious bleeding and necrosis,no fluid in the abdominal cavity.In the light microscope there was a significant pancreatic edema,a small amount of inflammatory cell infiltration and a very small amount of acinar cell necrosis,and no significant difference at each time point.6 The pancreas histopathological grading results: According to the Schmidt ’s methods for evaluation points,the control group at each time point does not change significantly in pathological damage score;the acute pancreatitis group each time point in the pancreas pathology injury score higher than the control group and the trimetazidine intervention group eachcorresponding point in time,there are significant difference(P<0.05);Trimetazidine intervention group pancreas at each time point histopathological injury score is higher than the control group each corresponding point in time,there are significant difference(P<0.05).7 Immunohistochemical demonstration: The control group pancreas tissue VEGF and i NOS at each time point are a few expression,score change is not obvious;the immunohistochemical score of the acute pancreatitis group pancreatic tissue VEGF and iNOS at each time point were higher than that in control group and the trimetazidine intervention group each corresponding point in time,there are significant difference(P<0.05);the immunohistochemical score of the trimetazidine intervention group pancreatic tissue VEGF and iNOS at each time point were lower than that in control group each corresponding point in time,there are significant difference(P<0.05).Conclusion:1 There is an increased in HIF-1 alpha,iNOS,NO,VEGF expression during acute pancreatitis.Increased expression of these factors plays an important role in the pancreas injury.2 The results suggest: trimetazidine on rat acute pancreatitis have a protective effect,its mechanism is: by inhibiting HIF-1alpha-iNOS-NO signaling pathways and HIF-1alpha-VEGF signaling pathways that control the inflammation reaction and microcirculation of acute pancreatitis.