Effect Of Anyi Granule On NF-?Bp65 And TXA₂/PGI₂ In Rats With Severe Acute Pancreatitis

Objective:To study the expression levels of Nuclear factor-kappa B p65?NF-?Bp65?,Cyclooxygenase-2?COX-2?,Thromboxane A2?TXA₂?,P rostacyclin?PGI₂?and TXA₂/PGI₂ratios in the pancreatic tissue of S evere Acute Pancreatitis?SAP?rats by Anyi Granules,and to investi gate the mechanism of Anyi Granule in the treatment of SAP to i mprove pancreatic microcirculation disorder.Methods:120 male adult Sprague-Dawley?SD?rats weighing 200g-250g were divided into four groups?30 rats in each group?according to the method of random number table,which were normal group,model group,western medicine group and Chinese medicine group.Each group was divided into 3 subgroups?10 rats in each subgroup?according to3h,6h and 12h,respectively,which were 3H group,6h group and 12h group.The rats in the normal group were free to drink and eat;The rats in the model group were fasted for 12 hours before modeling,and could not help drinking water.L-arginine solution?0.9%normal saline?with a concentration of 6%was injected into the abdominal cavity of rats at a dose of 150mg/100g,once an hour,three times in total,to induce sap;The rats in the western medicine group were injected with Recombinant human interleukin-10?rh IL-10?intraperitoneally 1 hour before modeling?10000U/each mouse?,and SAP was induced by the model group method.;The rats in the traditional Chinese medicine group were given Anyi Granules by gavage at a dose of 8g/kg?1g/ml?,once a day for three days,and SAP was induced by the model group method.Take the last SAP induction as the time point,and At 3h,6h and 12h,the rats were intraperitoneally injected with 1%Pentobarbital Sodium Solution?50mg/kg?in batches.After anesthesia,the rats were killed,and the pancreatic tissue was found.The pancreatic tissue was divided into two parts.One part was put into 4%formaldehyde solution,and labeled.It was used for the preparation of pathological sections and the detection of Integrated Optical Density?IOD?of NF-?Bp65 and COX-2 in imm-unohistochemistry;The other part uses Phosphate buffered saline?PBS?to remove the blood on the tissue surface,put it into EP tube of 0.9%Na Cl 1.5ml,after being treated by homogenizer,put it into centrifuge tube,mark it,put it in centrifuge,centrifuge it at 3000r/min for10min,take the serum,store it in refrigerator at-20?,and use enzyme linked immunosorbent assay?ELISA?to measure the TXA₂and PGI₂content of pancreatic tissue.statistical software package is applied.When the normal distribution is satisfied and the variances are uniform,the difference between the means is compared using a one-way analysis of variance.The LSD-t test is used to perform multiple comparisons of the means.When the normal distribution or variance is not satisfied,the rank sum test is used and multiple comparisons are made between groups.The inspection level is?=0.05.P<0.05 indicates that the difference is statistically significant.Results:?1?Histopathology of pancreas:in normal group,the structure of pancreatic lobules was normal,the acini,ducts and arrangement of lobules were regular,punctate nuclei were seen,and no pathological changes were found.In the model group,the structure of pancreatic lobule was disordered,inflammatory cell infiltration,intralobular duct dilation,acinar cell edema,scattered necrosis points,hemorrhage and necrosis of interstitial small vessel wall were observed,and the cell contour was fuzzy.In the western medicine group,the structure of the pancreatic lobule was intact,the acinar cells showed edema,and the interstitial microvascular wall showed different degrees of hemorrhage and necrosis,compared with the model group,the degree of pathological changes was lighter.In the traditional Chinese medicine group,the structure of the pancreatic lobule was slightly disordered,the lobule duct expanded,the acinar cells were edematous,and the swollen nucleus could be seen.Over time,local hemorrhage and necrosis could be seen in the acinus and stroma,compared with the model group,the degree of pathological changes was lighter.?2?IOD value of NF-?Bp65 in pancreas of rats in each group:Compared with the normal group,the IOD value of NF-?Bp65 in the other groups increased significantly at each time point?P<0.01?.Compared with the model group,the IOD value of NF-?Bp65 in the pancreas of the western medicine group and the traditional Chinese medicine group decreased significantly at each time point?P<0.01?.There was no statistically significant difference between the western medicine group and the Chinese medicine group at each time point?P>0.05?.?3?IOD value of COX-2 in pancreas of rats in each group:Compared with the normal group,the IOD value of COX-2 in the pancreatic tissues of other groups increased significantly at each time point?P<0.01?.Compared with the model group,the expression of IOD value of COX-2 in the pancreatic tissue of the western medicine group and the treatment group at each time point was significantly decreased?P<0.01?.There was no statistical significance between the western medicine group and the Chinese medicine group at each time point?P>0.05?.?4?The content of TXA₂and PGI₂in pancreas of rats in each group:Compared with the normal group,the content of TXA₂and PGI₂in the pancreas of the other groups increased significantly at each time point?P<0.01?;compared with the model group,the content of TXA₂and PGI₂in the pancreas of the western medicine group and the traditional Chinese medicine group decreased significantly?P<0.01?;Compared with the western medicine group,the contents of TXA₂and PGI₂in the pancreatic tissue of the traditional Chinese medicine group showed no statistically significant difference at each time point?P>0.05?.?5?Results of TXA₂/PGI₂ratio in pancreatic tissue of rats in each group:Compared with the normal group,the mean value of TXA₂/PGI₂ratio in the model group was significantly increased?P<0.01?.Compared with the model group,the mean value of TXA₂/PGI₂ratio of pancreatic tissue in the western medicine group and the traditional Chinese medicine group was significantly reduced?P<0.01?.There was no statistical significance between the western medicine group and the Chinese medicine group at each time point?P>0.05?.Conclusions:?1?Anyi granules can reduce pancreatic damage in rats with severe acute pancreatitis and protect the pancreas.?2?Anyi Granules can reduce the expression of NF-B and cox-2 in the pancreatic tissue of SAP rats,adjust the TXA₂/PGI₂ratio,reduce the damage caused by inflammation-induced microvessels,and then improve the microcirculation disorders of the pancreatic tissue,thereby protecting the pancreas.