| Objective Through injecting dibutyltin dichloride(DBTC)in caudal vein of SD rats established a model of chronic pancreatitis and pancreatic fibrosis,observing intervention on chronic pancreatitis fibrosis in rats from different concentrations of Ginkgo biloba extract(GBE),detecting changes of relevant indicators,so as to explore the mechanism of the prevention and treatment of pancreatic fibrosis.Methods 60 healthy male SD rats were randomly divided into 6 groups with 10 rats in each group.One group was recorded as Normal group.the rats of this group was injected the normal saline 1.5ml/kg into the tail vein;and the rats of the other five groups were injected DBTC1.5ml/kg into the tail vein,which were manufactured to the model of pancreatic fibrosis in rats with chronic pancreatitis.Observing the rats for two days after modeling,then the five groups were randomly marked as Model group,PTX group,GbE high dose group(200mg/kg),GbE middle dose group(100mg/kg),GbE low dose group(50mg/kg).Everyday,rats of GbE high,medium and low dose groups were administered GbE high,medium and low concentration suspension 10ml/kg;rats of Normal group and Model group were gavaged with equal volume of distilled water;however,rats of PTX group received intraperitoneal injection of PTX 6mg/kg,all rats were fed with standard rat feed and freely drinking water for four consecutive weeks.Four weeks later,all rats were anesthetized and the blood was taken from the abdominal cavity for serological detection;and then they were dissected for pancreatic tissue,which were fixed in 10%neutral formalin,paraffin embedded,sliced;hematoxylin eosin(HE)staining and collagen fiber Masson staining for observing the pancreatic tissue pathology;detecting the expression of Smad protein in pancreatic tissue by immunohistochemistry.Results(1)General situation and pancreatic tissues:rats of Normal group were generally good,no pancreatic injury;rats of Model group and GbE low dose group were generally poor,severe pancreatic tissue injury;The general situation of PTX group and GbE high and middle dose groups was better than that of Model group,pancreatic tissue damage was significantly lower than that of Model group and GbE low dose group.(2)Serological determination:compared with Normal group,the serum amylase and hyaluronic acid of Model group,PTX group and each GbE groups were higher(P<0.05),C-peptide were decreased significantly(P<0.01);however Compared with Model group,the serum amylase and hyaluronic acid of PTX group and GbE high,middle dose groups were lower(P<0.05),C-peptide were higher(P<0.05),which were significant differences with statistical significance.There was no significant difference between Model group and GbE low dose group(P>0.05),also no significant differences between each two of PTX group and GbE high or medium dose group(P>0.05).(3)HE staining:pancreatic tissue framework of Normal group was normal,Model group and GbE low dose group were significantly severe,the damage on pancreatic tissue of GbE high,middle dose groups and PTX group was less than that of Model group.(4)Masson staining:a small amount of collagen fibers were observed in the pancreatic duct and the blood vessels of Normal group;in Model group and GbE low dose group,a large number of collagen fibers were found in the pancreas,which were distributed in the small leaf,between the small leaf,in the catheter and aroud the blood vessels;compared with Model group,collagen fibers which were distributed in and between the small leaf was less than that in GbE high,middle dose groups and PTX group(P<0.05).Area ratio of collagen fiber in Normal group was significantly less than other groups(P<0.05),but there was no statistical difference between every two of GbE high,middle dose groups and PTX group(P>0.05).(5)Immunohistochemistry expression:①Smad2/3 protein:The expression of Smad2/3 protein in Model group,PTX group and GbE different dose groups was higher than that in Normal group(P<0.05);the expression of PTX group and GbE high.middle dose groups was lower than that of Model group(P<0.05).There was no significant difference between GbE low dose group and Model group(P>0.05),and no difference between every two of PTX group and GbE high,middle dose groups,too(P>0.05).②Smad7 protein:The expression of Smad7 protein in Model group,PTX group and GbE different dose groups was lower than that in Normal group(P<0.05);the expression of PTX group and GbE high,middle dose groups was higher than that of Model group(P<0.05).There was no significant difference between GbE low dose group and Model group(P>0.05),and no difference between PTX group and GbE high,middle dose groups,too(P>0.05).Conclusion The rat model of chronic pancreatitis with pancreatic fibrosis was successfully established by a single injection of DBTC in the rat tail vein.GbE had anti fibrosis effect on pancreatic fibrosis rats with chronic pancreatitis,and maybe was affected by its own concentration,the mechanism may be related to inhibiting the expression of Smad2/3 protein and enhancing the expression of Smad7 protein. |
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