Effect Of Probucol On Pancreatic Fibrosis In Rats

OBJECTIVE:To establish the rats model of chronic fibrosing pancreatitis and to prove the anti-fibrotic effect of probucol in chronic pancreatitis with fibrosis.METHODS:Part 1:48 male Sprague-Dawley Rats were put into 2 groups:congtrol group and model group,each group had 24 rats.Model group rats received intraperitoneal injection of 500mg/kg BW of DDC twice a week.Control group rats received intraperitoneal inje- ction of 0.5ml/kg BW of saline solution twice a week.From the end of the last intrap- eritoneal injection,4 rats of each group were killed at the 24hour ,48hour ,1week ,2w- eek,3 and 4 week. some of the pancreatic fragments were immediately fixed in 10% buffered formalin,processed for paraffin-embedded sectioning,and stained with hem- atoxylin and eosin.the results of microscopic finddings were scored by the pathologist with expertise in pancreatic pathology who was blinded to which experimental proce- dures were performed on the specimen under observation. Serum and panceraticlevels of Maleic Dialdehyde (MDA) and Superoxide Dismutase(SOD) were determined. Histopathological alterations were studied by optical microscopy. Expression of coll- agen was also examined while transforming growth factorbeta-1 (TGF-β₁),α-smooth muscle actin(α-SMA),Collagenâ… (Collagenâ… ) were localized by immunoche- mistry,fibrisis were scoring.Part 2 :Fifty rats were randomly divided into five groups, 10 rats in each group. Intr- aperitoneal injection of 500mg/kg BW of DDC twice a week.to induce chronic pancr- eatitis in rats (except fornormal group).probucol-treated rats were fed with different doses of probucol (75,150 and 300 mg/kg body weight) for 4 weeks, while normal group and control group received 0.9% sodium chloride solution. Serum and pancr- eatic levels of Maleic Dialdehyde (MDA) and Superoxide Dismutase(SOD) were de- termined. Histopathological alterations were studied by optical microscopy. Expre- ssion of collagen was also examined while transforming growth factor beta-1(TGF-β₁) were localized by immunochemistry,fibrisis were scoring.RESULTS :Part1:1. weight :In The first 1 weeks,model group rats grew slowly in contrast to control group(P>0.05).From the 1 week,rats in model group began to lose weight,there was a signiificant difference between the weight of the two group (P < 0.05);2.The expression of TGF-β₁,α-SMA,Collagenâ… : The expression of TGF-β₁,α-SMA,Collagenâ… were significantly higher than that of control group.Fibrosis appeared from the the first week in model group rats'pancreas,while there was little change in control group's.The difference was significant(P < 0.05).follow time passing,the The expression of TGF-β₁,α-SMA,Collagenâ… and Fibrosis were Rising as the 24hour<48 hour<1week<2 week<3 week<4 week.3. Serum and pancreatic levels of Maleic Dialdehyde (MDA) and Superoxide Dismutase(SOD): Serum and pancreatic levels of Maleic Dialdehyde (MDA) began to rise and Serum and pancreatic levels of Superoxide Dismutase(SOD) began to decline from the the first week in model group rats'pancreas,while there was little change in control group's.The difference was significant(P < 0.05).CONCLUTION 1:1.It takes 4 weeks to creat an animal model of chronic pancreatitis by intraperitoneal injection of 500mg/kg BW of DDC to male SD rats twice a week.2.pancreatic fibrosis and the expression of TGF-β₁were relation with oxidative Stress, oxidative Stress play a important role in the process of chronic pancreatitis. Part 21. weight :In The first 1 weeks, model group rats grew slowly in contrast to control group(P>0.05).after probucol treated, the chornic pancreatitis rats'weight there was rise ,there was a signiificant difference between the weight of the groups each other (P < 0.01);2. Fibrosis and the expression of TGF-β₁: while contrast with the model group,the Fibrosis and the expression of TGF-β₁ were less more.3. Maleic Dialdehyde (MDA): Serum and pancreatic levels of Maleic Dialdehyde (MDA) began to rise from the the model was induced in the model group rats',while there was contrary change in probucol-treatedl group's.The difference was significant(P < 0.01). described as model group(B group)>probucol low dose group(C group)>probucol middle dose group(D group)>probucol highe dose group(E group)>control group(A group).4. Superoxide Dismutase(SOD): Serum and pancreatic levels of and pancreatic levels of Superoxide Dismutase(SOD) began to decline from the the model was induced in the model group rats',while there was contrary change in probucol-treatedl group's.The difference was significant(P < 0.01) .described as model group(B group)1 .the correlation coefficient was : model group(B group)=0.860,P < 0.01;probucol low dose group(C group)=0.791,P < 0.05;probucol middle dose group(D group)=0.836,P < 0.05;probucol highe dose group(E group)=0.861,P < 0.01.6. Pancreatic MDA has a correlation with the extent of the expression of TGFβ₁ except probucol low dose group(C group).the correlation coefficient was : model group(B group)=0.718,P < 0.05;probucol low dose group(C group)=0.689,P > 0.05;probucol middle dose group(D group)=0.861,P < 0.01;probucol highe dose group(E group)=0.750,P < 0.05.7. Serum SOD has a correlation with the extent of the expression of TGFβ₁ .the correlation coefficient was : model group(B group)=-0.804,P < 0.05;probucol low dose group(C group)=-0.819,P < 0.05;probucol middle dose group(D group)=-0.805,P < 0.05;probucol highe dose group(E group)=-0.837,P < 0.01.8. Pancreatic SOD has a correlation with the extent of the expression of TGFβ₁ except probucol low dose group(C group).the correlation coefficient was : model group(B group)=-0.737,P < 0.05;probucol low dose group(C group)=-0.687,P > 0.05;probucol middle dose group(D group)=-0.781,P < 0.05;probucol highe dose group(E group)=-0.846,P < 0.01.9. CONCLUTION 2:Probucol has an anti-fibrotic effect on pancreatic fibrosis in rats. Because of its Elimination oxygen free radical effect and the expression of TGFβ₁ significant.it could be a potential drug for the treatment of chronic pancreatitis.