Effect Of The Treatment Of Smoothing Liver Tonifying Spleen And Dredging Collaterals On Expression Of Smad2/3 And Smad7 Of The Rats With Chronic Pancreatitis And Pancreatic Fibrosis

Objective To establish a model of chronic pancreatitis and pancreatic fibrosis in Spraque-Dawley(SD)rats by tail intravenous disposable injection of dibutyltin dichloride(DBTC).Observing the effect of the treatment of smoothing liver tonifying spleen and dredging collaterals on rats pancreatic tissue fibrosis and its impact on the degree of relevant indicators,and to explore the possible mechanism of the prevention and treatment of chronic pancreatitis and pancreatic fibrosis.Methods 40 healthy male SD rats were randomly divided into the normal control group 10 which to be disposable tail vein injection saline(1.5ml/kg),and the other 30 rats were prepared for the pancreatic fibrosis model group,which injected via the tail vein of the same volume of DBTC(1.5ml/kg).Observed two days later,and then the survived pancreatic fibrosis rats were randomly divided into the model group,the TCM group and the pentoxifylline(PTX)group.After grouping,the TCM group were fed traditional Chinese medicine(10ml/kg)every day,the normal group and the model group were fed the same volume of distilled water(10ml/kg)every day,the PTX group were injected the PTX solution 6mg(kg/d).After four weeks,anesthetized the rats and drew blood from the abdominal aorta for serologic test,took the pancreatic tissue and fixed in 10%neutral formalin solution,embedded in paraffin,sections for hematoxylin-eosin(HE)staining,Masson staining and immunohistochemistry for histological observation and diagnosis.Results(1)the general conditions and the rats pancreatic tissues in general:the rats of the normal control group were generally good,the pancreatic tissue injury was not obvious;The model group were generally poor and the pancreatic tissue injury was more serious;the TCM group and PTX group were better than the model group,the pancreatic tissue injury was significantly reduced.(2)Determination of serum Hyaluronic acid and C-peptide:Compared with the normal control group,the serum hyaluronic acid of the model group,the TCM group and the PTX was significantly higher(P<0.05),the C-peptide was Significantly lower(P<0.05).When compared with the model group,the serum hyaluronic acid level of the TCM group and the PTX was reduced(P<0.05),C-peptide was increased(P<0.05).But the serological markers between the TCM group and the PTX group had no significant difference(P>0.05).(3)HE staining and pathological score:The basic structure of pancreatic tissue of the normal control group were normal,the model group were damaged severely,the TCM group and the PTX group were better than the model group.The pathological scores of the model group,the TCM group and the PTX group was significantly higher than the normal control group(P<0.05),the TCM group and the PTX group were lower than the model group(P<0.05),but the pathological scores between the TCM group and the PTX group had no significant difference(P>0.05).(4)Masson staining and determination of fibrous area:The normal control group showed that a small amount of fibrous distributed in the pancreatic duct tissue and perivascular,while in the model group there were a lot of fibrous distributing in interlobular and intralobular pancreas.The fibrous area of the TCM group and the PTX group were less than that in the model group(P<0.05),but that between the TCM group and the PTX group had no significant difference(P>0.05).(5)Immunohistochemical expression of Smad2/3 and Smad7:The pancreatic tissue expression of Smad2/3 in the model group,the TCM group and PTX group were higher than the normal control group(P<0.05).Compared with the model group,the TCM group and the PTX group were decreased(P<0.05).but there were no significant difference between the TCM group and the PTX group(P>0.05).The pancreatic tissue expression of Smad7 in the normal control group were higher than the other groups(P<0.05).Drug intervention groups were higher than the model group(P<0.05),but there were no significant difference between the TCM group and the PTX group(P>0.05).Conclusion(1)The model of the chronic pancreatitisand pancreatic fibrosis can be successfully established by disposable rat tail vein injection DBTC.(2)The treatment of smoothing liver tonifying spleen and dredging collaterals can change pathology of the pancreatic fibrosis,reduce the levels of serum hyaluronic acid and increase the levels of C-peptide.(3)It can play a role of anti-fibrosis in rats with chronic pancreatitis and pancreatic fibrosis through inhibiting the expression of Smad2/3 and promoting the expression of Smad7.