Experimental Study On The Effect Of Anyi Granule On NF-?Bp65 And ET/NO In Severe Acute Pancreatitis Rats

Objective: Based on the nuclear factor-?B(NF-?B)signaling pathway,explore the effects of Anyi granules on the expression of iNOS,the levels of ET and NO,and the ET/NO ratio,clarify the mechanism of Anyi granules in treating severe acute pancreatitis and improve microcirculation disorder,and provide experimental basis for clinical treatment of severe acute pancreatitis.Methods:The experimental animals were SD male rats,clean grade,weighing about 200-250 g,and randomly divided into normal control group,SAP(model group)group,IL-10(western medicine group)group,and Anyi granules(Chinese medicine group).)Group consists of four large groups with 30 SD rats in each group;each group is divided into 3h,6h and 12 h groups,with 10 SD rats in each subgroup.The rats in the SAP group were fasted and water-free for12 hours before modeling,and were intraperitoneally injected with L-Arginine solution,150mg/100 g,once per hour for three times,inducing severe acute pancreatitis.The IL-10 group was intraperitoneally injected with 10000 U rh IL-10 before pre-modeling(the same method as SAP group);The Anyi granule group was given intragastric administration of Anyi granules(8g/kg,equivalent to 5 times the clinical daily recommended dose)3 days before modeling,once a day for a total of 3 days after modeling(the same method as SAP group);the normal control group had a normaldiet.The time node was the last intraperitoneal injection in the SAP group.Rats in each group were sacrificed by 1% pentobarbital(50mg/kg)anesthesia at 3h,6h,and 12 h,and pancreatic tissue was harvested.A part of pancreas tissue was fixed in 4% formaldehyde solution for pathological observation and immunohistochemical method to detect the expression of NF-?Bp65 and i NOS in pancreas tissue;The other part of pancreas tissue was homogenized by adding appropriate amount of PBS solution,and then centrifuged quickly at3000r/min.The supernatant was taken and stored in a refrigerator at-20? for ELISA to detect ET and NO levels in pancreatic tissue.Results:(1)Pathological results show:Pancreas tissue slices of normal control group showed intact pancreatic lobular structure,and very few slices showed a small amount of neutrophil infiltration;The SAP group showed obvious edema between the pancreatic lobules,the structure of the lobules was obscured,a large number of neutrophils were infiltrated,the acinar cells had extensive necrosis,the interstitial small blood vessel wall hemorrhage and necrosis,and the red blood cell overflow;The structural integrity of pancreatic tissues in the pancreatic tissues of the Anyi granule group and IL-10 group and the degree of edema between the leaflets,necrosis and neutrophil infiltration were all reduced compared with SAP group.(2)Immunohistochemical results show:(1)Compared with the normal group,the expression of NF-?Bp65 in the SAP group was significantly increased(P<0.01);compared with the SAP group,the expression of NF-?Bp65 in the IL-10 group and Anyi granule groupwas significantly reduced(P<0.01),There was no significant difference between Anyi Granule group and IL-10 group(P>0.05).(2)Compared with the normal group,the i NOS expression in the SAP group increased significantly(P<0.01);compared with the SAP group,the expression of i NOS in IL-10 and Anyi granule groups were significantly reduced(P<0.01),but there was no statistically significant difference between Anyi granule group and IL-10 group(P>0.05)(3)ELISA results show:Compared with the normal group,the content of ET and NO in the SAP group increased significantly(P<0.01);Compared with the SAP group,the ET and NO contents of the IL-10 group and the Anyi granule group were significantly reduced(P<0.01),but compared with the IL-10 group,the ET and NO contents of the Anyi granule were not Statistical significance(P>0.05),(4)ET/NO ratio: Compared with the normal group,the ET/NO ratio of the SAP group was significantly increased(P<0.01);compared with the SAP group,the ET/NO ratio in the IL-10 group and Anyi granule group was significantly reduced(P<0.01),but there was no statistically significant difference between the Anyi granule group and the IL-10 group(P> 0.05).Conclusions: 1.Anyi granules can reduce the pathological damage of severe acute pancreatitis rats and protect the pancreatic tissue;2.Anyi granules can improve the pancreatic microcirculation by inhibiting the NF-?B signal pathway,thereby suppressing the expression of i NOS,reducing the content of vascular active substances ET and NO,correcting the imbalance of ET/NO,andimproving the microcirculation of the pancreas,so as to achieve the effect of treating severe acute pancreatitis.