| Objective: To investigate the effects of Anyi Granule on nuclear factor ?Bp65(NF-?Bp65),inducible nitric oxide synthetase(iNOS),cyclo oxygenase-2(COX-2),tumor necrosis factor alpha(TNF-?),interleukin-1?(IL-1?)in rats with severe acute pancreatitis induced by L-arginine,to clarify the main mechanism of Anyi granule in the treatment of severe acute pancreatitis,to enrich the clinical treatment of severe acute pancreatitis,and to improve the treatment of severe acute pancreatitis Prognosis of pancreatitis.Methods: 120 Sprague Dawley(SD)rats were randomly divided into 4 groups by computer random number table after 7 days of adaptive feeding.The rats were divided into normal group,model group,traditional Chinese medicine group and Western medicine group.Each group was further divided into three groups: 3 hours,6hours and 12 hours,with 10 rats in each group.In the traditional Chinese medicine group,8g/kg anyi granules were dissolved in water for three consecutive days before modeling,and then gavage was performed once a day.Normal group did not do treatment,free water diet;Western medicine group 1 hour before building be Recombinant human interleukin 10(rh IL-10)10000 u by intraperitoneal injection of line;After the above pretreatment,rats in the model group,traditional Chinese medicine group and westernmedicine group were fed with water for 12 hours before modeling,and injected with 6% l-arginine solution at a dose of 150mg/100 g intraperitoneally for 3 times,each time at an interval of 1 hour,to induce SAP model.From the end time of the last injection,abdominal aortic blood was taken from the four groups of rats by groups of 3 hours,6 hours and 12 hours after anesthesia,and the rats were sacrificed for anatomy,the gross morphology of the pancreas was observed,and pancreatic tissue was rapidly extracted.Serum TNF-?,IL-1? levels were measured by enzyme-linked immunosorbent assay.The pancreatic tissue was taken and the pathological sections were made.The contents of NF-?Bp65mRNA were detected by real-time PCR,and the expressions of NF-?Bp65,iNOS and COX-2 were determined by immunohistochemistry.All data were collected for statistical analysis.Results:(1)The results of pathological section of pancreas showed that the structure of pancreatic lobule was clear,the morphological structure of acinar cell,glandular duct and islet was normal,and the dark blue punctate nucleus was seen in acinar cell.In the model group,the structure of lobule was fuzzy,the arrangement of cells was disordered,the broken cells piled up in pieces,the visual field was fuzzy,the acinar cells were necrotic in large area,the nucleus swam out or broke up,the gap was widened,the inflammatory cells infiltrated,the interstitial small blood vessels were broken and necrotic,and the red blood cells overflowed.Compared with the model group,the structure of pancreatic lobule in the traditional Chinese medicine group was still visible,the inflammatory cell infiltration,vascular rupture,cell necrosis were significantlyimproved,the cell swelling was obvious,and the cell outline and nucleus in place were still visible.The changes of pancreas in western medicine group were similar to those in traditional Chinese medicine group.(2)Serum TNF-? and IL-1?: TNF-? and IL-1? levels increased significantly with the time extension in all the other groups except the normal group.12 small time points were the peaks in the observation time of this experiment,and there were statistically significant differences(TNF-? p < 0.01,IL-1? p < 0.05).Compared with the model group at the same time point,the serum TNF-? and IL-1? of the traditional Chinese medicine group and the western medicine group decreased significantly,with statistically significant difference(p<0.01).There was no statistically significant difference between the traditional Chinese medicine group and the western medicine group(p > 0.05).(3)NF-?Bp65 mRNA in pancreatic tissue: Except for the blank group,NF-?Bp65 mRNA in pancreatic tissue of other groups increased significantly with time extension at each time point after modeling,and the peak value at 12 hours was the observation time of this experiment,and there was statistical significance between each time point and the blank group(P <0.01);compared with the model group at each time point,NF-?Bp65 in pancreatic tissue of the treatment group and Western medicine group NF-?Bp65 mRNA decreased significantly(P <0.01).There was no significant difference between the treatment group and the western medicine group(P > 0.05).(4)NF-?Bp65,iNOS and COX-2 in pancreatic tissue: Except for the normal group,NF-?Bp65,iNOS,and COX-2 in the pancreatic tissues of the other groups increased significantly with time extension at each time point after modeling.Twelve small time points were the peaks in the observation time of this experiment,and the difference between the same time point and the normal group was statistically significant(p <0.01).Compared with the model group at the same time point,NF-?Bp65,iNOS and COX-2in the pancreatic tissues of the traditional Chinese medicine group and the western medicine group decreased significantly(p <0.01).There was no statistically significant difference between the traditional Chinese medicine group and the western medicine group(p >0.05).(5)The expression of NF-?Bp65 in the pancreas of SAP rats was highly positively correlated with the expression of TNF-?,IL-1?,iNOS,and COX-2(p < 0.05),and the positive correlation was also found between TNF-?,IL-1?,iNOS,and COX-2(p <0.05).Conclusion: 1.The levels of TNF-?,IL-1 ?,NF-?Bp65mRNA,NF-?Bp65,iNOS and COX-2 in SAP type serum and pancreas tissue induced by L-arginine began to increase significantly at the time point of 3 hours,and reached the peak at 12 hours in this group of experiments.NF-?Bp65 was highly positively correlated with the expression of each factor,and there was also a positive correlation between each factor in varying degrees,indicating that NF-?Bp65mRNA gene transcription induction The expression of TNF-?,IL-1 ?,NF-?Bp65,iNOS and COX-2 was detected at a certain level.Inflammatory amplification and microcirculationdisturbance occurred simultaneously in the early stage of sap,and inflammatory factors and microcirculation affected each other.2.Anyi granule can effectively improve the pathological damage of pancreatic tissue in SAP rats,which shows that Anyi granule has a protective effect on pancreatic tissue and is the pathophysiological basis of effective treatment of SAP.3.After gastric perfusion,Anyi granule not only significantly reduced the content of TNF-?and IL-1 ? in serum of SAP rats,but also significantly reduced the expression of NF-?Bp65 mRNA and NF-?Bp65,iNOS and COX-2in pancreatic tissue,suggesting that Anyi granule could prevent and treat SAP systemic inflammatory response syndrome through nuclear factor inflammatory factor pathway,improve microcirculation through nuclear factor microcirculation factor pathway,and effectively reduce the inflammation in the process of SAP onset Sexual injury and microcirculation disturbance can effectively treat sap. |
PDF Full Text Request