ABSTRACT
PURPOSE: This study aimed to elucidate the molecular mechanisms of the anti-pancreatic fibrosis effects of matrine in rats. MATERIALS AND METHODS: Trinitrobenzene sulfonic acid was administrated to rats to establish a pancreatic fibrosis model. Rats were divided into four groups: Control, Sham, Model, and Matrine (n=8). Hematoxylin-eosin staining, Masson staining, and Azan staining were performed to evaluate pancreatic fibrosis. Expression of transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), and collagen I in pancreatic tissues was evaluated by immunohistochemical staining. mRNA and protein levels of TGF-β receptor 1 (TβR1), TβR2, and Smad2 in pancreatic tissues were determined by RT-PCR and Western blot, respectively. RESULTS: In the model group, hyperplasia of glandules around the glandular ducts, mitochondrial swelling of acinous cells, and severe fibrosis were found. Interestingly, in the Matrine group, mitochondrial swelling was only found in a small number of acinous cells, and the fundamental structures of pancreatic tissues were intact. Moreover, pancreatic fibrosis was markedly alleviated. Comparing to the Sham group, expression of α-SMA, TGF-β1, and collagen I was sharply elevated in the Model group (p < 0.05); however, their expressions were much lower in the Matrine group, compared to the Model group (p < 0.05). Compared with the Sham group, mRNA and protein levels of Smad2, TβR1, and TβR2 in the Model group were notably raised (p < 0.05). However, their high expression was significantly downregulated in the Matrine group (p < 0.05). CONCLUSION: Matrine suppressed pancreatic fibrosis by regulating TGF-β/Smad signaling in rats.
Subject(s)
Animals , Rats , Acinar Cells , Actins , Blotting, Western , Collagen , Fibrosis , Hyperplasia , Mitochondrial Swelling , RNA, Messenger , Signal TransductionABSTRACT
PURPOSE: A soft texture of the pancreas is one of the most important predisposing factors for a pancreatic fistula. Thus, in a porcine model, we investigated a method to harden the pancreas locally by directly injecting an artificial material. METHODS: During the laparotomy, 51 samples from 17 pigs, including 13 survival models, were randomly divided into 3 groups and either received a direct injection into the pancreas of MHL (1:4 mixture of histoacryl [n-butyl cyanoacrylate] and lipiodol) (group E) or saline (group C) or only received a pinprick into the pancreas without injecting a substance (sham). We measured the change in the pancreatic hardness after the injection using a durometer and examined the histological change of the pancreas using the fibrosis grade in the survival model. RESULTS: The postinjection hardness of the pancreas was significantly increased in group E compared to group C and the sham group (P < 0.001). Pathologically, all cases in group E showed a severe fibrotic change, whereas the other groups demonstrated mild to no fibrosis (P < 0.001). The fibrosis in group E was localized to the area of the injection, while the surrounding areas were preserved. CONCLUSION: The direct injection of MHL could induce focal hardening and fibrotic changes in the pancreas of the porcine model.
Subject(s)
Causality , Cyanoacrylates , Enbucrilate , Ethiodized Oil , Fibrosis , Hardness , Laparotomy , Methods , Pancreas , Pancreatic Fistula , SwineABSTRACT
Objective: To investigate the effect of oxymatrine (OM) on the expression of Gli2 in LTC-14 cells and PANC-1 cells induced by TGF-β1. Methods: LTC-14 cells and PANC-1 cells were divided into four groups:control group, TGF-β1 group, TGF-β1 + OM group, and OM group. The protein was extracted after 12 h, and the expressions of related proteins were detected by Western blot. Results: Compared with control group, the expression of Gli2 in LTC-14 cells was significantly decreased induced by TGF-β1, while the expressions of α-SMA, FN, and CoL-I were significantly increased. The expressions of Gli2, FN, CoL-I, and α-SMA were increased significantly induced by TGF-β1when compared with those of control group in PANC-1 cells, and the expression of Gli2 was inhibited by OM pretreatment in TGF-β1 + OM group compared with TGF-β1 group. OM pretreatment can increase the expression of Gli2 before stimulating with TGF-β1 in TGF-β1 + OM group in comparison with TGF-β1 group. Conclusion: OM plays an protected role in pancreatic fibrosis through promoting the expression of Gli2 in LTC-14 cells.
ABSTRACT
Objective To investigate the risk factors of postoperative pancreatic fistula after pancreaticoduodenectomy (PD).Methods The retrospective case-control study was adopted.The clinicopathological data of 196 patients with PD who were admitted to First Affiliated Hospital of Dalian Medical University from September 2014 to July 2016 were collected.All the patients underwent PD.Observation indicators:(1) intra-and postoperative situations;(2) follow-up;(3) analysis of risk factors of pancreatic fistula after PD.All patients were followed up by outpatient examination and telephone interview to detect pancreatic fistula and peritoneal fluid collection up to March 2017.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed by t test.Measurement data with skewed distribution were represented as median (range).Count data and univariate analysis were done using the chi-square test.Logistic regression model was used for multivariate analysis.Results (1) Intra-and post-operative situations:all the 196 patients underwent surgeries successfully.The operation time,volume of intraoperative blood loss,number of intraoperative blood transfusion and non intraoperative blood transfusion were (439± 136) minutes,(686±280) mL,45 and 151 cases,respectively.Time to initial anal exsufflation,time of initial defecation and time for first diet after operation were (4.1 ±2.1) days,(5.1± 2.9) days and (3.1 ± 2.0) days.Of 76 patients,the content of diastase in the i ntraperitoneal drainage was 614 U/L (31-30 215 U/L) at postoperative day 1 and level of serum procalciton in was (0.7±0.4) ng/mL at postoperative day 3.Time for drainage tube removal of 196 patients was (14.6±7.1)days.Fifty four of 196 patients with postoperative complications were improved by symptomatic treatment,including 15 with intestinal obstruction,12 with delayed gastric emptying,11 with abdominal infection,9 with incision infection,7 with bleeding.Duration of postoperative hospital stay was (17.1 ±4.2)days.Results of pathological diagnosis of 196 patients showed 121 cases of pancreatic cancer,50 of intraductal papillary mucinous tumors of the pancreas,7ampullary carcinoma,15 of carcinoma of the lower end of the bile duct,and 3 of duodenum cancer.Pancreatic findings:pancreatic texture:95 cases were with soft pancreas and 101 with hard pancreas.Diameter of main pancreatic duct duct:101 cases had diameter of pancreatic duct duct ≥3 mm and 95 cases ＜3 mm.(2)Followup:all the 196 patients were followed up for 4-30 months,with a median follow-up time of 18 months.During follow-up time,the grade B/or C pancreatic fistula occurred in 37 cases.Of 16 patients with pancreatic fistularalated ascites,10 had readmission and were improved by symptomatic treatment.(3) Analysis of risk factors of pancreatic fistula after PD:the results of univariate analysis showed that the content of diastase in the intraperitoneal drainage at postoperative day 1,level of serum procalcitonin at postoperative day 3 and pancreatic texture were related factors affecting the pancreatic fistula after PD (x2 =6.569,5.902,13.517,P＜0.05).The results of multivariate analysis showed that the content of diastase in the intraperitoneal drainage at postoperative day 1 ≥600 U/L was an independent risk factor affecting the pancreatic fistula after PD (OR =9.135,95％confidence interval:2.247-37.130,P＜0.05).Conclusion The content of diastase in the intraperitoneal drainage at postoperative day 1 ≥ 600 U/L is an independent risk factor affecting the pancreatic fistula after PD.
ABSTRACT
Objective To investigate the risk factors of postoperative pancreatic fistula after pancreaticoduodenectomy (PD).Methods The retrospective case-control study was adopted.The clinicopathological data of 196 patients with PD who were admitted to First Affiliated Hospital of Dalian Medical University from September 2014 to July 2016 were collected.All the patients underwent PD.Observation indicators:(1) intra-and postoperative situations;(2) follow-up;(3) analysis of risk factors of pancreatic fistula after PD.All patients were followed up by outpatient examination and telephone interview to detect pancreatic fistula and peritoneal fluid collection up to March 2017.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed by t test.Measurement data with skewed distribution were represented as median (range).Count data and univariate analysis were done using the chi-square test.Logistic regression model was used for multivariate analysis.Results (1) Intra-and post-operative situations:all the 196 patients underwent surgeries successfully.The operation time,volume of intraoperative blood loss,number of intraoperative blood transfusion and non intraoperative blood transfusion were (439± 136) minutes,(686±280) mL,45 and 151 cases,respectively.Time to initial anal exsufflation,time of initial defecation and time for first diet after operation were (4.1 ±2.1) days,(5.1± 2.9) days and (3.1 ± 2.0) days.Of 76 patients,the content of diastase in the i ntraperitoneal drainage was 614 U/L (31-30 215 U/L) at postoperative day 1 and level of serum procalciton in was (0.7±0.4) ng/mL at postoperative day 3.Time for drainage tube removal of 196 patients was (14.6±7.1)days.Fifty four of 196 patients with postoperative complications were improved by symptomatic treatment,including 15 with intestinal obstruction,12 with delayed gastric emptying,11 with abdominal infection,9 with incision infection,7 with bleeding.Duration of postoperative hospital stay was (17.1 ±4.2)days.Results of pathological diagnosis of 196 patients showed 121 cases of pancreatic cancer,50 of intraductal papillary mucinous tumors of the pancreas,7ampullary carcinoma,15 of carcinoma of the lower end of the bile duct,and 3 of duodenum cancer.Pancreatic findings:pancreatic texture:95 cases were with soft pancreas and 101 with hard pancreas.Diameter of main pancreatic duct duct:101 cases had diameter of pancreatic duct duct ≥3 mm and 95 cases ＜3 mm.(2)Followup:all the 196 patients were followed up for 4-30 months,with a median follow-up time of 18 months.During follow-up time,the grade B/or C pancreatic fistula occurred in 37 cases.Of 16 patients with pancreatic fistularalated ascites,10 had readmission and were improved by symptomatic treatment.(3) Analysis of risk factors of pancreatic fistula after PD:the results of univariate analysis showed that the content of diastase in the intraperitoneal drainage at postoperative day 1,level of serum procalcitonin at postoperative day 3 and pancreatic texture were related factors affecting the pancreatic fistula after PD (x2 =6.569,5.902,13.517,P＜0.05).The results of multivariate analysis showed that the content of diastase in the intraperitoneal drainage at postoperative day 1 ≥600 U/L was an independent risk factor affecting the pancreatic fistula after PD (OR =9.135,95％confidence interval:2.247-37.130,P＜0.05).Conclusion The content of diastase in the intraperitoneal drainage at postoperative day 1 ≥ 600 U/L is an independent risk factor affecting the pancreatic fistula after PD.
ABSTRACT
Objective: To investigate the effect of oxymatrine (OM) on expression of related molecules in Smad3, Gli1 signaling pathway in PANC-1 cells induced by TGF-β1. Methods: TGF-β1-induced PANC-1 cells were used to establish the pancreatic fibrosis model in vitro, and observe the effects of OM pretreatment on the related molecular expression of Smad3/Gli1. Gli1 and Smad3 RNA interference plasmids were transfected into PANC-1 cells. The protein expression levels of Smad3, Gli1 and α-SMA were measured by Western blotting. The levels of fibronectin (FN) and type I collagen (CoL-I) in the supernatant of cell culture were detected by ELISA. Results: Compared with the control group, the protein expressions of Smad3, Gli1 and α-SMA increased significantly in PANC-1 cells after treated with TGF-β1. The expressions of Gli1, α-SMA, FN, and CoL-I in PANC-1 cells decreased significantly after Gli1 RNA interference plasmid transfection compared with TGF-β1 induced group. The expression of Smad3, Gli1, α-SMA, FN, and CoL-I also decreased significantly in PANC-1 cells after Smad3 RNA interference plasmid transfection compared with TGF-β1 group. Conclusion: OM could prevent pancreatic fibrosis by regulating TGF-β1/Smad3/Gli1 signaling pathway in PANC-1 cells.
ABSTRACT
[ ABSTRACT] AIM:To explore the role of superoxide dismutase ( SOD) and malondialdehyde ( MDA) in chronic pancreatitis ( CP) induced by dibutyltin dichloride ( DBTC) combined with ethanol, and the mechanisms for prevention and treatment of pancreatic fibrosis by Chaihushugansan.METHODS: The KM mice were randomly divided into control group, CP group ( DBTC combined with ethanol) and Chaihushugansan group ( CP+Chaihushugansan) .Except for control group, the mice in other groups were intravenously injected in tail with DBTC (8 mg/kg) and drank 10% ethanol.The mice in Chaihushugansan group were administered intragastrically with Chaihushugansan (6 g· kg-1 · d-1 ) at the follow-ing experimenal period.Before modeling and 1 week, 2 weeks, 4 weeks and 8 weeks after modeling, the mice were anes-thetized and sacrificed.The activity of amylase and the content of hyaluronic acid in the serum were measured.The mor-phology and the degree of fibrosis in the pancreas were observed by HE staining.The activity of SOD and the level of MDA in the pancreas homogenate were analyzed.The protein of pancreas was extracted to detect the expression of type I collagen by Western blotting.RESULTS:DBTC combined with ethanol induced CP with increased serum amylase and hyaluronic acid levels, while the serum amylase and hyaluronic acid levels in Chaihushugansan group were significantly lowered ( P<0.05).In 1 week, 2 weeks, 4 weeks and 8 weeks, the pancreas were obviously injured and appeared different degrees of fibrosis.The content of MDA and the expression of type I collagen in the increased significantly, but the SOD was de-creased.In Chaihushugansan group, the pathological damage and the degree of fibrosis of the pancreas were improved.The level of MDA and type I collagen expression in the pancreas were significantly reduced, but the SOD was increased.CON-CLUSION:The oxidative stress may take part in the development of CP.Inhibition of oxidative stress in the pancreas is one of the mechanisms that Chaihushugansan attenuates the development of CP.
ABSTRACT
Objective To observe the effect of Xiaotan Hezhong Recipe on pancreatic fibrosis forming and the expression of related proteins in model rats of chronic pancreatitis. Methods Twenty-four Wistar rats were randomly divided into normal group, model group, pancreatin group and Chinese medicine group (n=6). Rats model were made by DDC solution intraperitoneal injection. Pancreatin group and Chinese medicine group were treated by intragastric administration with pancreatin and Xiaotan Hezhong Recipe, respectively. Body weight of rats was measured regularly. The rats were killed after 4 weeks, with pancreatic pathological detection and laminin, matrix metalloproteinase-1 detection. Results After intraperitoneal injection of DDC solution, pancreatic tissue fibrosis in different degree and inflammatory cell infiltration were found. Chinese medicine group was better than the model group in general conditions, body weight change, pancreatic pathological score and expression of laminin, matrix metalloproteinase-1, the differences were statistically significant (P<0.05). Conclusion Xiaotan Hezhong Recipe can improve chronic pancreatitis rats generally and inhibit pancreatic fibrosis, its mechanism may be related with decreasing laminin expression and increasing the expression of matrix metalloproteinase-1.
ABSTRACT
BACKGROUND/AIMS: High-fat diets contribute to pancreatic fibrogenesis, but the pathogenesis remains unclear. This study investigated the role of nuclear factor kappa B (NF-kappaB) in high-fat diet-induced pancreatic fibrosis in rats. METHODS: Male Wistar rats were fed a high-fat diet or standard normal chow for 20 weeks. Pancreatic fibrosis was determined by Sirius red staining. Immunohistochemical staining, reverse transcription-polymerase chain reaction and Western blotting were used to identify NF-kappaB-associated genes or protein expressions. RESULTS: Inflammation, fat deposition, pancreatic stellate cell activation and fibrosis were observed in the pancreases of the high-fat diet group. NF-kappaB subunit p65 (NF-kappaB/p65) expression was localized to the nucleus, and intercellular adhesion molecule 1 (ICAM-1) was over-expressed. Pancreatic gene expression levels of NF-kappaB/p65, ICAM-1 and tumor necrosis factor alpha were all elevated significantly in rats fed a high-fat diet compared with control rats. Western blotting also revealed significantly increased levels of ICAM-1 and nuclear NF-kappaB/p65 in rats fed high-fat diets comparison with control rats. CONCLUSIONS: NF-kappaB is involved in high-fat diet-related pancreatic fibrosis.
Subject(s)
Animals , Humans , Male , Rats , Blotting, Western , Diet, High-Fat , Fibrosis , Gene Expression , Inflammation , Intercellular Adhesion Molecule-1 , NF-kappa B , Pancreas , Pancreatic Stellate Cells , Rats, Wistar , Tumor Necrosis Factor-alphaABSTRACT
Objective To evaluate the diagnostic effect of intro-operative tissue puncture biopsy and find its influential factors.Methods The clinical data of 94 patients with pancreatic mass treated in our hospital from July 1994 to December 2007 and undergoing intro-operative tissue puncture biopsy were retrospectively analyzed.Results The sensitivity,the specificity,the positive predictive value and the negative predictive value were 74.6%,93.8%,98.0%and 46.9%,respectively.The single factor analysis showed that the size of pancreatic mass,the number of puncture and complicating with pancreatic fibrosis or not were related to the diagnosis of tissue puncture.The multiple factor analysis showed that the size of pancreatic mass(P=0.014)and the number of puncture(P=0.020)were crucial to the diagnosis of puncture.The sensitivity and specificity of intro-operative tissue puncture biopsy for the pancreatic mass less than 25 mm were lower than that for the mass larger than 25 mm (P=0.000).The sensitivity and specificity would be increased as the number of puncture increased (P=0.000).For the mass less than 25 mm,increasing the humber of puncture would improve the sensitivity(P=0.002).Conclusion Intro-operative tissue puncture biopsy is a simple and accurate procedure for differentiating the pancreatic mass.The sensitivity and specificity could be improved by increasing the number of puncture,especially for the patients with pancreatic mass less than 25 mm.
ABSTRACT
Objective To introduce the role of pancreatic stellate cells in pancreatic fibrosis and the progress in treatment of pancreatic fibrosis. Methods Relevant literatures were collected and reviewed. Results Pancreatic stellate cells activation was closely related to pancreatic fibrosis. Inhibition of pancreatic stellate cells activation could provide a new approach in clinical treatment of chronic pancreatitis. Conclusion Pancreatic stellate cells are the key to pancreatic fibrosis,which are becoming the target for anti-fibrosis of the pancreas and treatment of chronic pancreatitis.
ABSTRACT
AIM To investigate the effects and possible mechanism s of Glycyrrhizin on rat pancreatic fibrosis induced by TNBS (trinitrobenzenesulfonic acid, TNBS ). METHODS Chronic pancreatitis model was induced in male Sprague -Dawley rats by injection of 2% TNBS into bile duct. All the rats were randomly divided into two groups. The rats in Glycyrrhizin intervention group were treat ed with Glycyrrhizin 8 mg?kg -1 by injection into tail vein from day 3 to day 28, while the rats in control group were administrated with same volume of saline vehicle. Ten rats in the Glycyrrhizin intervention group and eight rats in the control group wer e sacrificed on day 29, the blood was collected to determine amylase and hyaluro nic acid by enzyme dynamic and RIA method. The histological change of pancreatic tissue was evaluated by H&E stain and modified Van-Gieson stain. Mast cell in pancreas was stained by thionine blue. Expression of TGF-? 1,Collagen Ⅰ and ?-SMA in pancreas were assessed by immunohistochemistry and western blot. RESULTS In the Glycyrrhizin intervention group, the mast cell number and the percentage of degranulation decreased significantly, and the expression of ?-SMA protein also decreased compared to the control group, but there was no difference in amylase or hyaluronic acid between the treatment group and the control group. In the Glycyrrhizin intervention group, inflammation and fibrosis were ameliorated and expression of collageⅠ and TGF-? 1 was also decreased significantly compared to the control group. CONCLUSION Glycyrrhizin inhibits pancreatic fibrosis in chronic pancreatitis rats induced by TNBS. This action might be related to protecting pancreatic acinus cells from being destructed by mast cell activation and inhibiting extracellular matrix synthesis stimulated by pancreatic stellate cell.
ABSTRACT
Objectives To analyze the role and mechanisms of mast cells in the inflammation and fibrosis of 2 ,4, 6-trinitrobenzene sulfonic acid (TNBS)-induced rat pancreatic fibrosis. Methods Rats were received the aseptic instillation of TNBS in ethanol via bilo-pancreatic duct, and then injected with nedocromil sodium, a mast cell stabilizer, and compound 18/80, a mast cell activator, or saline. Rats were sacrificed respectively on 3, 7, 14, 21 or 28 days. Pancreatic inflammation and fibrosis were assessed by gross and histopathological evaluation. Pancreatic fibrosis were observed by Van Gieson. Pancreatic mast cells distribution, number and their state of activation (toluidine blue) were evaluated. The activation of pancreatic stellate cells (PSCs) were assessed by the expression of a-smooth muscle actin (?-SMA) through immunohistochemistry. The expression of angiotensin Ⅱ AT1 and AT2 receptors and transforming growth factor (TGF) ? 1 raRNA, which were the factors of fibrogenesis, were also assessed. Results Typical pancreatic fibrosis changes occurred in the model of rats received TNBS at 4th week by morphological evaluation. The positive expression of ?-SMA and TGF?1 in the pancreatic tissues were detected at day 3, especially at 4th week. The expression of angiotensin Ⅱ AT1 and AT2 receptors mRNA increased gradually in all the three groups, also especially at 4th week. Compared to the control group, there were more higher expression of ?-SMA, TGF?1, angiotensin Ⅱ AT1 and AT2 receptor in the compound 48/80 group, while there were lower expression of these proteins in the nedocromil group. Conclusions Mast cells are involved in TNBS-induced pancreatic inflammation and fibrosis in rats. After being activated, mast cells will promote the activation and proliferation of PSCs and upregulate the expression of angiotensin Ⅱ AT1 receptor and AT2 receptor, and then lead to pancreatic fibrosis gradually.
ABSTRACT
Objective To observe the activation of pancreatic stellate cells (PSC) during the formation of pancreatic fibrosis induced by the pancreatic injection of trinitrobenzene sulfonic acid (TNBS). Meanwhile, the effects of PSC-related factors, such as transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 on the pathogenesis of pancreatic fibrosis in rats were also evaluated. Methods Pancreatic fibrosis model in rats was induced by the injection of 2% TNBS in ethanolate-phosphate buffer solution into the pancreatic duct. The rats were sacrificed and the pancreata were removed at the 72nd hour, 3rd week, 4th week, 5th week, 6th week and 7th week after the operation respectively. Expressions of ?-smooth muscle actin (?-SMA), transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 were determined by either immunohistochemistry or RT-PCR, or Western blot respectively. The ultrastructure of pancreas was studied by electron microscope at different time points. Results The inflammation, swelling and necrosis were the major pathological changes of the pancreas at the early stage after the injection of 2% TNBS. Subsequently, the fibrotic manifestations such as proliferation of the fibrosis, atrophy of vesicles, deposition of collagen because prominent at the 3rd week after the operation, which peaked at 4th week. The expression of TGF-? 1 was increased significantly at the 3rd week after the operation and reached maximum at the 4th week. The expression of ?-SMA, which indicated the activation of PSC, could be detected at the 3rd week and also reached the peak value at the 4th week. After wards, it was decreased gradually. During the first 72 hours, the expression of MMP-2 mRNA was increased significantly and then was fluctuated but still higher than that in normal rats. The deposition of type Ⅰ collagen was increased in the areas of fibrotic tissues. Conclusions PSC might involve in the courses of the development and progression of TNBS induced pancreatic fibrosis in rats. This action was achieved via the activation of PSC by TGF-? 1, the production of those extracellular matrix metabolic associated enzymes such as the synthesis of collagen Ⅰ and the secretion of MMP-2.