Pancreatic Histological Changes Research Articles

Frequency of detection and load of amastigotes in the pancreas of Leishmania infantum-seropositive dogs: clinical signs and histological changes

BackgroundZoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and is highly lethal in humans and dogs if left untreated. The frequency of this parasite and associated histological changes in the pancreas of dogs are poorly studied. Therefore, the objectives of this study were to evaluate the frequency of detection and load of amastigotes in the pancreas of L. infantum-seropositive dogs and to identify the clinical signs and histological changes associated with parasitism of this organ.MethodsOne hundred forty-three dogs from an endemic area in Brazil that tested seropositive for L. infantum were studied. The dogs were clinically examined, killed, and necropsied between 2013 and 2014. One fragment of the pancreas was randomly collected for histopathology and immunohistochemistry, and spleen and bone marrow were collected for culture.ResultsLeishmania amastigotes were detected in the pancreas of 22 dogs (15.4%) by immunohistochemistry, all exhibiting L. infantum parasitism in the spleen and/or bone marrow. Poor body condition and cachexia were only associated with infection of the pancreas with Leishmania spp. (p = 0.021) and were found in 40.9% of dogs with pancreatic infection. Anorexia, vomiting, and/or diarrhea were observed in 9.2% of dogs with pancreatitis. The median parasite load in the pancreas was 1.4 infected macrophages/mm2. Pancreatic histological changes and their frequencies were: granulomatous pancreatitis (28.0%), lymphoplasmacytic pancreatitis (23.8%), acinar cell degeneration (6.3%), fibrosis (5.6%), hemorrhage (2.1%), eosinophilic pancreatitis (0.7%), suppurative pancreatitis (0.7%), and necrosis (0.7%).ConclusionsThe present results demonstrate that L. infantum is one of the etiological agents of chronic pancreatitis in dogs; however, the frequency of detection and parasite load are low in this organ. The lack of an association of poor body condition and cachexia with pancreatitis and the low frequency of clinical signs commonly associated with pancreatitis suggest that a significant portion of the organ is not affected by this parasite. On the other hand, the association of poor body condition and cachexia with concomitant infection of the pancreas, spleen, and/or bone marrow with this parasite suggests that these manifestations are the result of a more advanced stage of canine visceral leishmaniasis.Graphic abstract

The Pancreatic Histological Changes of Morizella® Juice

Background Morizella juice is a nutritional supplement juice prepared from Moringa oleifera leaves and Hibiscus sabdariffa (HS) calyces. The juice is believed to have stabilizing effect on blood pressure (BP), anxiety, and to control glucose levels in some cases of diabetes. However, it is not recommended for consumption by infants and people with low blood pressure. Broad objective The aim of the current study was to evaluate the pancreatic histological changes in rats fed with Morizella juice. Materials and Methods A total of 18 healthy male and female Wistar rats, weighting between 183 and 224mg of 2-3 months old were used. The experiments were divided into short-term, mid-term and long-term test of six animals per group. Animals were sacrificed, the pancreatic tissues were harvested and histologically processed. Sections were examined and photographed using light microscope with in-built camera, and the number of Islet cells were measured in 40 high-power fields. Results Microscopic investigation of the pancreatic tissue in both treated groups revealed morphological changes in hormone-secreting endocrine tissues as evidenced by putative proliferation of Islets of Langerhans when compared with the control group. The number of Islet cells was significantly higher (p < 0.05) in all treated groups at the dosages of 175 and 350 mg/kg body weight. It is important to note that our results also showed the increase in number of intra-islets capillaries in all treated rats. Conclusions The beneficial effect of Morizella juice was observed in rats in all doses studied herein. It was found to increase the number of pancreatic endocrine cells as evidenced by pancreatic tissue proliferation with significant increase in intra-islet vascularization. Recommendations Further research aiming at the evaluation of histological changes of Morizella juice by using special stains for detection of the proliferation of islet cells and to recognize the ?-cells of islets is recommended. Keywords: Morizella juice, Moringa oleifera, Proliferation, Hibiscus sabdariffa, Tanzania.

Pancreatic histological changes in adult female albino rats treated with Orlistat and the possible protective role of B- carotene.

Orlistat is a pancreatic lipase inhibitor licensed for obesity treatment. It is considered to be a safe drug for long term use but many adverse effects were observed such as pancreatitis. B-carotene is a precursor of vit. A which has antioxidant, anti-inflammatory and immune-enhancing effects. Objective: This study aims to clarify the pancreatic structural changes after Orlistat administration and the possible protective role of B- carotene in adult female albino rats.Materials & Methods: Fifty adult female albino rats were divided into four groups: Group I (10rats) was control. Group II (10rats) was administered B-carotene at a dose of 0.52 mg/kg/day, orally for 5 weeks. Group III(20rats) was administered a therapeutic dose of Orlistat (32 mg/kg/day) dissolved in 1 ml distilled water orally for 5 weeks then 10 rats were scarified(subgroup III a) and the others 10 rats left without intervention for another 5 weeks(subgroup III b). Group IV (10 rats) was administered B-carotene as group II one hour before the administration of orlistat as group III a for 5 weeks. At the end of the study, each rat was weighed, blood samples were obtained for estimation of blood glucose level and pancreas were dissected and prepared for histological study.Results: Orlistat treated rats showed a highly significant decrease in body weight and a highly significant increase in blood glucose level. There were signs of B cells inhibition in anti-insulin immunostaining. Destructive changes of acinar cells with marked reduction of secretory granules and strong positive reaction for iNOS reactions were observed. These changes were confirmed by electron microscopic examination. These changes were reversible in withdrawal group. B- carotene coadministration showed amelioration of the histological changes.Conclusion:B- carotene supplementation has ameliorating effect on pancreas against the deleterious effects of orlistat.

TThe effect of Atorvastatin on the pancreas of adult male albino rats and the possible protective role of Resveratrol (Histological, Immunohistochemical and Biochemical study)

Introduction: Hyperlipidaemia is a dangerous condition that may lead to atherosclerosis and various cardiovascular diseases. Different drugs are taken to reduce hyperlipidaemia such as Statins. Recently various side effects for statins have been detected clinically that actually restrict their prescription. Resveratrol is a natural phenol and is produced naturally by several plants and proved to have both antioxidant & anti-inflammatory actions.Aim of the Study: This research has been aimed to investigate the effect of Atorvastatin on the pancreas and the probable protective action of Resveratrol.Materials and Methods: Forty adult male albino rats were divided into four equal groups and were received drugs orally as a single daily dose for one week. Control group (I): received 1ml normal saline. Resveratrol group (II): received dose 30 mg/kg. Atorvastatin group (III): received dose 80 mg/Kg. Group IV: received Resveratrol concomitant with Atorvastatin with the same doses as pervious groups.All rats were sacrificed one day following the last dose and blood samples were gathered for biochemical study of serum amylase and lipase. Pancreas specimens were obtained and processed for histological and immunohistochemical study.Results: Atorvastatin-treated rats showed degeneration of pancreatic acini. The acinar cells showed cytoplasmic vacuoles with dilated RER and depleted zymogen granules. Also, inflammatory cell infiltration, dilated congested blood vessels with hemorrhage and marked deposition of collagen fibers were observed. The islets of Langerhans showed apparent reduced cells number. α cells and to more extent β cells revealed degeneration. The immunohistochemical results revealed marked increase in vascular endothelial growth factor (VEGF) and caspase-3 expressions. The biochemical study revealed remarkable elevation in the serum amylase and lipase. The concomitant administration of Resveratrol with Atorvastatin minimizes these changes. Conclusion: Atorvastatin has been proved to induce pancreatic histological changes and these changes can be attenuated by Resveratrol when given concomitantly with it.

Fucoxanthin ameliorates hyperglycemia, hyperlipidemia and insulin resistance in diabetic mice partially through IRS-1/PI3K/Akt and AMPK pathways

Fucoxanthin has potential anti-diabetic activities, but its precise effects and underlying mechanisms remain unclear. In this study, C57BL/KsJ-db/db mice were fed diets containing fucoxanthin (0.2% and 0.4%, w/w) for 6 weeks. Results showed dietary fucoxanthin significantly improved glucose/lipid metabolism and insulin resistance, and prevented pancreatic histological changes in db/db mice. Moreover, fucoxanthin markedly lowered blood glucose by regulating glucokinase and phosphoenolpyruvate carboxykinase mRNA expressions in liver, promoted glycogen synthesis by regulating glucose transporter 4, glycogen synthase and glycogen synthase kinase-3 beta protein expressions in skeletal muscle, and lowered blood lipid levels by regulating peroxisome proliferator-activated receptor alpha, p-acetyl CoA carboxylase, carnitine palmitoyltransferase 1 and fatty acid synthetase protein expressions in liver. Mechanistic studies indicated that fucoxanthin effectively regulates the expression of IRS-1/PI3K/AKT and AMPK signaling proteins in liver and skeletal muscle. All results suggested that fucoxanthin is a potential functional food or effective drug for type 2 diabetes mellitus.

Combination of Obestatin and Bone Marrow Mesenchymal Stem Cells Prevents Aggravation of Endocrine Pancreatic Damage in Type II Diabetic Rats.

One of the new promising therapies in treatment of diabetes mellitus is mesenchymal stem cells (MSCs) which have an interesting therapeutic potentiality based on their paracrine effect and transdifferentiation potentiality. Also obestatin improves the generation of functional β cells/islet-like cell clusters in vitro, suggesting implications for cell-based replacement therapy in diabetes. So the aim of this study was to evaluate the effect of combination of both MSCs and obestatin on an experimental model of type II diabetes mellitus (T2DM). Sixty male rats were divided into; group I (control group), group II (T2DM group) induced by administration of high fat diet (HFD) and injection of streptozotocin (STZ) in low dose, group III (T2DM treated with MSCs), group IV (T2DM treated with obestatin), group V (T2DM treated with MSCs and obestatin). Fasting blood glucose, C-peptide, insulin and lipid profile were measured. HOMA-IR and HOMA-β were calculated. Pancreatic expression of insulin, glucagon like peptide -1 (GLP-1) and pancreatic duodenal homeobox 1 (Pdx1) mRNA levels were measured. In addition pancreatic histological changes, insulin and Bax were analyzed by immunohistochemical examination of islets of Langerhans. Diabetic rats showed significant increase in HOMA-IR, serum glucose and lipid profile levels with significant decrease in insulin, HOMA-β, GLP-1 and Pdx1 levels. MSCs and obestatin caused significant improvement in all parameters with more significant improvement in combined therapy. The protective effects afforded by MSCs and obestatin may derive from improvement of the metabolic profile, antiapoptosis and by increase in pancreatic GLP-1and Pdx1 gene expression.

Jiang Tang Xiao Ke Granule Play an Anti-diabetic Role in Diabetic Mice Pancreatic Tissue by Regulating the mRNAs and MicroRNAs Associated with PI3K-Akt Signaling Pathway.

Purpose: To investigate the effect of JTXK granule on the expression pattern of miRNA in pancreatic tissue of KKAy diabetic mice, and to explore the molecular mechanism and pathways of JTXK granule in anti-diabetic effect.Methods: We used high fat diet (HFD) to induce the KKAy diabetic mice and screened the differentially expressed miRNAs (DEMs) between JTXK-treated group (n = 6) and the diabetic group (n = 6) using MicroRNA (miRNA) Microarray. C57BL/6J mice were given a normal diet as the control group (n = 6). Subsequently, miRNA target gene prediction, GO and Pathway analysis were used to explore the function of DEMs. Finally, the mechanism of anti-diabetic effects of JTXK granule was tested by in vitro INS-1 pancreatic β-cell experiment.Results: The blood glucose and body weight of JTXK-treated group was significantly lower compared with the model group. Moreover, a total of 45 miRNAs with significant differences were detected in the model group and the JTXK-treated group (P ≤ 0.05, Fold Change > 2). Further, miRNA-mRNA analysis showed that the differential expression of mmu-miR-192-5p, mmu-miR-291a-3p, mmu-miR-320-3p, mmu-miR-139-5p and mmu-miR-378a-3p are closely related to pancreatic histological changes. In addition, pathway analysis showed that the DEMs were closely related to PI3K-Akt Signaling Pathway. Furthermore, the levels of serine/threonine-protein kinase (Akt), phosphorylated Akt (p-Akt) and phosphorylated forkhead transcription factor O1 (p-Foxo1) in INS-1-FOXO1 overexpressing model cells were lower than those in normal group, while JTXK granules could increase the expression of Akt, p-Akt and p-Foxo1.Conclusions: The results showed that JTXK granule could play an anti-diabetic role by regulating the mRNA and miRNAs associated with PI3K-Akt pathway in diabetic mice pancreatic tissue.

Changes in pancreatic histology, insulin secretion and oxidative status in diabetic rats following treatment with Ficus deltoidea and vitexin

BackgroundThe potential application of Ficus deltoidea and vitexin for the management of symptomatologies associated with diabetes mellitus (DM) has gained much attention. However, less firm evidence comes from data to augment our understanding of the role of F. deltoidea and vitexin in protecting pancreatic β-cells. The aim of this study was to assess histological and oxidative stress changes in the pancreas of streptozotocin (STZ)-induced diabetic rats following F. deltoidea extract and vitexin treatment.MethodsF. deltoidea and vitexin was administrated orally to six-weeks STZ-induced diabetic rats over 8 weeks period. The glucose and insulin tolerances were assessed by intraperitoneal glucose (2 g/kg) tolerance test (IPGTT) and intraperitoneal insulin (0.65 U/kg) tolerance test (IPITT), respectively. Subsequently, insulin resistance was assessed by homeostasis assessment model of insulin resistance (HOMA-IR), quantitative insulin sensitivity check index (QUICKI) and the insulin/triglyceride-derived McAuley index. The histological changes in the pancreas were then observed by hematoxylin-eosin (H&E) staining. Further, the pattern of fatty acid composition and infrared (IR) spectra of the serum and pancreas were monitored by gas chromatography (GC) method and Fourier Transform Infrared (FT-IR) spectroscopy.ResultsF. deltoidea and vitexin increased pancreatic antioxidant enzymes and promoted islet regeneration. However, a significant increase in insulin secretion was observed only in rats treated with F. deltoidea. More importantly, reduction of fasting blood glucose is consistent with reduced FT-IR peaks at 1200-1000 cm−1.ConclusionsThese results accentuate that F. deltoidea and vitexin could be a potential agent to attenuate pancreatic oxidative damage and advocate their therapeutic potential for treating DM.

Incretin Therapies Do Not Expand β-Cell Mass or Alter Pancreatic Histology in Young Male Mice.

The impact of incretins upon pancreatic β-cell expansion remains extremely controversial. Multiple studies indicate that incretin-based therapies can increase β-cell proliferation, and incretins have been hypothesized to expand β-cell mass. However, disagreement exists on whether incretins increase β-cell mass. Moreover, some reports indicate that incretins may cause metaplastic changes in pancreatic histology. To resolve these questions, we treated a large cohort of mice with incretin-based therapies and carried out a rigorous analysis of β-cell turnover and pancreatic histology using high-throughput imaging. Young mice received exenatide via osmotic pump, des-fluoro-sitagliptin, or glipizide compounded in diet for 2 weeks (short-term) on a low-fat diet (LFD) or 4.5 months (long-term) on a LFD or high-fat diet (HFD). Pancreata were quantified for β-cell turnover and mass. Slides were examined for gross anatomical and microscopic changes to exocrine pancreas. Short-term des-fluoro-sitagliptin increased serum insulin and induced modest β-cell proliferation but no change in β-cell mass. Long-term incretin therapy in HFD-fed mice resulted in reduced weight gain, improved glucose homeostasis, and abrogated β-cell mass expansion. No evidence for rapidly dividing progenitor cells was found in islets or pancreatic parenchyma, indicating that incretins do not induce islet neogenesis or pancreatic metaplasia. Contrasting prior reports, we found no evidence of β-cell mass expansion after acute or chronic incretin therapy. Chronic incretin administration was not associated with histological abnormalities in pancreatic parenchyma; mice did not develop tumors, pancreatitis, or ductal hyperplasia. We conclude that incretin therapies do not generate β-cells or alter pancreatic histology in young mice.

Evaluation of fish oil-rich in MUFAs for anti-diabetic and anti-inflammation potential in experimental type 2 diabetic rats.

The advantages of monounsaturated fatty acids (MUFAs) on insulin resistance and type 2 diabetes mellitus (T2DM) have been well established. However, the molecular mechanisms of the anti-diabetic action of MUFAs remain unclear. This study examined the anti-hyperglycemic effect and explored the molecular mechanisms involved in the actions of fish oil- rich in MUFAs that had been acquired from hybrid catfish (Pangasius larnaudii×Pangasianodon hypophthalmus) among experimental type 2 diabetic rats. Diabetic rats that were fed with fish oil (500 and 1,000 mg/kg BW) for 12 weeks significantly reduced the fasting plasma glucose levels without increasing the plasma insulin levels. The diminishing levels of plasma lipids and the muscle triglyceride accumulation as well as the plasma leptin levels were identified in T2DM rats, which had been administrated with fish oil. Notably, the plasma adiponectin levels increased among these rats. The fish oil supplementation also improved glucose tolerance, insulin sensitivity and pancreatic histological changes. Moreover, the supplementation of fish oil improved insulin signaling (p-AktSer473 and p-PKC-ζ/λThr410/403), p-AMPKThr172 and membrane GLUT4 protein expressions, whereas the protein expressions of pro-inflammatory cytokines (TNF-α and nuclear NF-κB) as well as p-PKC-θThr538 were down regulated in the skeletal muscle. These data indicate that the effects of fish oil-rich in MUFAs in these T2DM rats were partly due to the attenuation of insulin resistance and an improvement in the adipokine imbalance. The mechanisms of the anti-hyperglycemic effect are involved in the improvement of insulin signaling, AMPK activation, GLUT4 translocation and suppression of pro-inflammatory cytokine protein expressions.

Blockade of Multidrug Resistance-Associated Proteins Aggravates Acute Pancreatitis and Blunts Atrial Natriuretic Factor's Beneficial Effect in Rats: Role of MRP4 (ABCC4).

We previously reported that atrial natriuretic factor (ANF) stimulates secretin-evoked cAMP efflux through multidrug resistance-associated protein 4 (MRP4) in the exocrine pancreas. Here we sought to establish in vivo whether this mechanism was involved in acute pancreatitis onset in the rat. Rats pretreated with or without probenecid (MRPs general inhibitor) were infused with secretin alone or with ANF. A set of these animals were given repetitive cerulein injections to induce acute pancreatitis. Plasma amylase and intrapancreatic trypsin activities were measured and histological examination of the pancreas performed. Secretin alone activated trypsinogen but induced no pancreatic histological changes. Blockade by probenecid in secretin-treated rats increased trypsin and also induced vacuolization, a hallmark of acute pancreatitis. ANF prevented the secretin response but in the absence of probenecid. In rats with acute pancreatitis, pretreatment with secretin aggravated the disease, but ANF prevented secretin-induced changes. Blockade of MRPs in rats with acute pancreatitis induced trypsinogen activation and larger cytoplasmic vacuoles as well as larger areas of necrosis and edema that were aggravated by secretin but not prevented by ANF. The temporal resolution of intracellular cAMP levels seems critical in the onset of acute pancreatitis, since secretin-evoked cAMP in a context of MRP inhibition makes the pancreas prone to injury in normal rats and aggravates the onset of acute pancreatitis. Present findings support a protective role for ANF mediated by cAMP extrusion through MRP4 and further suggest that the regulation of MRP4 by ANF would be relevant to maintain pancreatic acinar cell homeostasis.

Pancreatic fatty degeneration and fibrosis as predisposing factors for the development of pancreatic ductal adenocarcinoma.

Knowledge of risk factors for development of pancreatic ductal adenocarcinoma (PDAC) is limited. To clarify the background condition of the pancreas for the development of PDAC, we analyzed pancreatic histological changes in noncancerous lesion specimens after pancreatectomy in PDAC patients. Seventy-six patients with PDAC were enrolled in this study. The PDAC was in the pancreatic head in 37 patients, in the body in 31, and in the tail in 8. No patients had a history of clinical chronic pancreatitis. As controls, 98 patients without PDAC were enrolled. The following parameters were examined: fibrosis, fatty degeneration, and inflammatory cell infiltration. More than 5% of fatty degeneration in the specimen, more than 10% of fibrosis, and more than 5% of inflammatory cell infiltration were considered positive changes. Pancreatectomy specimens showed a higher ratio of positive change in fibrosis (86% vs 42%), fatty degeneration (72% vs 44%), and inflammatory cell infiltration (14% vs 3%) than control samples. Multivariate analyses demonstrated that each histological change was a significant, independent determinant for PDAC. Our study demonstrated that cryptogenic pancreatic inflammation with fatty changes represents an important predisposing factor for PDAC. Screening for subclinical chronic pancreatitis in healthy populations may enable the detection of PDAC at an early stage.

Abstract 244: SPINK1 insufficiency induces impaired autophagy resulting in chronic pancreatitis

Abstract Background&amp; Objective: Several studies reported that chronic pancreatitis is a risk factor for pancreatic ductal adenocarcinoma (PDAC). Serine protease inhibitor Kazal type 1 (SPINK1), also known as pancreatic secretory trypsin inhibitor (PSTI), was originally isolated from the pancreas. In mice, the homologous gene is designated as Spink3. SPINK1/Spink3 covalently can bind to activated trypsin within the pancreatic acinar cells to form an inactive and stable complex and to prevent acute pancreatitis, a major inflammatory disorder of the pancreas. The mutations of SPINK1 gene are associated with various forms of idiopathic pancreatitis. We previously showed that deletion of Spink3 causes pancreatitis-like changes in the mouse. A careful molecular and pathological analysis of this genetically engineered mouse model will reveal the chronic of nflammation in the pancreas. The aim of this study was to rescue the Spink3-/- phenotype by generating Spink3-/- mice with knocked-in SPINK1, and to analyze molecular mechanisms that initiate pancreatitis or drive its progression from acute to chronic disease in this genetic model. Methods: We placed SPINK1 minigene (KI) into mouse Spink3 locus, using the Cre-loxP technology and generated chimeric mice (Spink3KI/+). Spink3KI/+ mice were crossed to Spink3+/- mice, thus generating Spink3KI/-. Mice were followed up to 8 weeks old. Time-dependent changes in pancreatic histology, stellate cell ctivation, and serum amylase were measured. Results: Spink3KI/KI mice showed no abnormalities in pancreas or any other organ during the 8 weeks age of observation, indicating that human SPINK1 can rescue Spink3 deficiency in physiological condition. Spink3KI/- mice showed slight growth etardation. The pancreas of Spink3KI/- mice at birth contained normal acinar cells. The Spink3KI/- mice time-dependently developed pathologic features of chronic tissue damage, including loss of acinar cells, intralobular fibrosis with activated pancreatic stellate cells. Interlobular ducts were dilated and contained protein plugs, which resembled chronic pancreatitis in human. In the pancreas of Spink3KI/- mice, we found that trypsin activity increased, and large vacuoles, derived from autophagy, appeared in acinar cells. p62, which is a selective substrate for autophagy aggregated in acinar cells, suggested that autophagy is impaired in Spink3KI/- acinar cells. Conclusions: We demonstrated that SPINK1 insufficiency caused chronic pancreatitis in vivo. Impaired autophagy and high trypsin activity may contribute to cause chronic pancreatitis in human.We established chronic pancreatitis model in human SPINK1 knockin mice and identified shared pathological alterations in human and mouse chronic pancreatitis.Thus,this mouse model provides a suitable tool to analyze the oncogenesis from chronic pancreatitis. Citation Format: Kazuya Sakata, Masaki Ohmuraya, Katsunobu Taki, Daisuke Hashimoto, Satoshi Ida, Hidetoshi Nitta, Hiromitu Hayashi, Akira Chikamoto, Tooru Beppu, Hideo Baba. SPINK1 insufficiency induces impaired autophagy resulting in chronic pancreatitis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 244. doi:10.1158/1538-7445.AM2014-244

Therapeutic effect of sodium aescinate on severe acute pancreatitis in rats

Objective To investigate the therapeutic effect of sodium aescinate (SA) on severe acute pancreatitis (SAP) in rats.Methods SAP model was induced by intraductal administration of 5％ taurocholic acid sodium salt.SD rats were then randomly divided into 3 groups (n =16),namely sham,model and SA group (5 mg/kg).Drugs or saline(10 ml/kg) were administered via caudal vein 30 min post-operation.The serum amylase was measured with spectrophotometer and pancreatic histological changes were observed under microscope 6 h and 12 h post-operation,respectiveiy.Results Pancreatic histological grades in the model group were (9.42 ± 1.06) and (18.30± 2.18) 6 h and 12 h post-operation,respectively,which showed significant differences (P＜0.01) compared with the sham group [(0.55 ±0.20) and 0,respectively].SA evidently reduced the severity of pancreatic pathology and improved the tissue inflammation in rats with SAP.Pancreatic histological grades were (7.85±1.33) and (12.75±1.69)in the SA group 6 h and 12 h post-operation,respectively,which had significant differences compared with the model group (P＜0.05).Serum amylase levels in model group were (2612 ± 59) U/L and(3004±687)U/L6 h and 12 h post-operation,respectively,which showed significant differences (P ＜ 0.01) compared with the sham group [(928 ± 271) and (890± 295) U/L,respectively].Serum amylase levels were reduced to (1790± 336) U/L and (2093 ± 298) U/L in the SA group 6 h and 12 h post-operation,respectively.SA markedly inhibited the level of serum amylase compared to the model group (P＜0.01).Conclusion SA could protect rats with SAP effectively. Key words: Sodium aescinate; Severe acute pancreatitis; Rats

Abstract 5188: Chronic pancreatitis induces the cancer-related gene expressions .

Abstract Background &amp; Aims: Several studies reported that chronic pancreatitis (CP) is a risk factor for pancreatic ductal adenocarcinoma (PDAC). The loss-of-function mutations of serine protease inhibitor Kazal type 1 (SPINK1) gene are associated with various forms of hereditary pancreatitis.We previously showed that deletion of Spink3,the mouse homologue of human SPINK1, causes pancreatitis-like changes in the mouse. A careful molecular and pathological analysis of evolving PDAC has revealed a characteristic pattern of genetic lesions. The field is now faced with the challenge of understanding how these signature genetic lesions _ mutations of cancer-related genes _ contribute to the biological characteristics and evolution of this disease. The aim of this study was to rescue the Spink3−/- phenotype by generating Spink3−/- mice with knocked-in SPINK1, and to characterize time-dependent changes of cancer-related genes in the pancreas in this genetic model. Methods: We placed CAG promoter-SPINK1 minigene(SP1) into diaphanous homolog 2 (Diap2) locus, which is located on X chromosome, using the Cre-Lox technology. X-inactivation is a process whereby one of the two copies of the X chromosome present in female mammals is inactivated. By utilizing X-inactivation,we were able to create mice in which SPINK level was partially, but not completely, reduced compared with the wild type. The SP1 knock-in mice(male YXSP1 and female XSP1/SP1 or XSP1/+) were crossed to Spink3+/- mice, thus generating Spink3−/−SP1 knock-in mice. Mice were followed up for up to 6 months.Time-dependent changes in pancreatic histology, stellate cell activation, and serum amylase were measured. Results: Spink3−/−YXSP1,as well as Spink3−/−XSP1/SP1 mice in which SP1 transgene was present on both X chromosomes, showed no abnormalities in pancreas or any other organ during the 6 months of observation, indicating that human Spink1 rescues Spink3 deficiency in mice. Spink3−/−XSP1/+ mice in which SP1 transgene is present on only one of the two X chromosomes, showed slight growth retardation but were healthy and fertile. Pancreas of Spink3−/−XSP1/+ mice at birth contained both normal and degenerated acinar cells, with accumulation of autophagic vacuoles. The Spink3−/−XSP1/+ mice time-dependently developed pathologic features of chronic tissue damage, including loss of acinar cells, intralobular fibrosis with activated stellate cells. Interlobular ducts were dilated and contained protein plugs, which resembled chronic pancreatitis in human. Older mice displayed acinar-ductal metaplasia and prominent expression of proto-oncogenes Egfr, Her2, and Ras. Conclusions: CP may contribute to carcinogenesis by preventing the senescence barrier characteristic of normal pancreas. Antiinflammatory treatment of CP patients may reduce their risk of developing PDAC. Citation Format: Kazuya Sakata, Masaki Ohmuraya, Daisuke Hashimoto, Hidetoshi Nitta, Katunori Imai, Hiromitu Hayashi, Yoshiaki Ikuta, Akira Chikamoto, Tooru Beppu, Hideo Baba. Chronic pancreatitis induces the cancer-related gene expressions . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5188. doi:10.1158/1538-7445.AM2013-5188

Inflexinol Reduces Severity of Acute Pancreatitis by Inhibiting Nuclear Factor-κB Activation in Cerulein-Induced Pancreatitis

To examine the effect of inflexinol on the development of acute pancreatitis (AP) and to investigate the mechanisms responsible for the protective effect against AP. Acute pancreatitis was induced in mice by intraperitoneal injection of cerulein. Inflexinol was administered intraperitoneally 4 times every 6 hours from 1 hour before the first cerulein injection. Serum amylase activity and histology of the pancreas were measured. Determination of pancreatic nuclear factor-κB (NF-κB) p65 expression was conducted by Western blotting and immunohistochemistry to investigate the mechanisms responsible for the inflexinol effects. Serum amylase activity in the cerulein group was significantly higher than that in the control group (P < 0.05). Pancreatic histology revealed marked inflammatory changes in the cerulein group such as interstitial edema, vacuolization, necrosis, and infiltration of inflammatory cells; and Western blotting and immunohistochemistry showed marked NF-κB p65 expression. Treatment with inflexinol significantly attenuated the inflammatory changes in pancreatic histology at 24, 48, and 72 hours (P < 0.05). Pancreatic NF-κB p65 expression decreased significantly after inflexinol treatment (P < 0.05). Inflexinol reduced the severity of cerulein-induced AP by inhibiting NF-κB activation.

Studies in rodents with the dipeptidyl peptidase‐4 inhibitor vildagliptin to evaluate possible drug‐induced pancreatic histological changes that are predictive of pancreatitis and cancer development in man

The present report summarizes rodent studies with vildagliptin, relevant to predicting pancreatitis or pancreatic cancer in man. As part of the regulatory development program for vildagliptin, a rodent toxicity program included two 104-week rodent (mouse and rat) carcinogenicity studies that were conducted according to guidelines assigned in Food and Drug Administration's Draft Guidance for Industry. Vildagliptin exposure in animals was evaluated for its effects on endocrine and exocrine pancreas. Two-year carcinogenicity studies were conducted in rats at oral doses up to 900 mg/kg (approximately 200 times the human exposure at the maximum recommended dose) and in mice at oral doses up to 1000 mg/kg (up to 240 times the human exposure at the maximum recommended dose). The results from these studies show the expected preservation and growth of the endocrine β-cells with no significant findings in the exocrine acinar pancreas. There was no evidence of inflammatory infiltrates characteristic of pancreatitis, no palpable mass detection based on gross examination or any microscopic findings indicative of pancreatic islet cell (endocrine), acinar cell (exocrine) or ductal (exocrine) neoplasia in rat or mouse. Evaluation of vildagliptin in 2-year preclinical carcinogenicity studies in both rats and mice indicates that while vildagliptin results in pharmacological benefits to the endocrine pancreas, this was not associated with any evidence of pancreatitis, pancreatic islet cell, acinar cell or ductal neoplasia. These data predict no increased risk of pancreatic cancer in man.

Su1237 Pancreatic Enzyme Supplementation for Treatment of Pain in Adult Patients With Chronic Pancreatitis: A Meta-Analysis and Systematic Review

Background & Aims: Chronic pancreatitis (CP) is characterized by chronic inflammation and progressive pancreatic fibrosis leading eventually to the loss of exocrine and endocrine pancreas functions. The loss-of-function mutations of serine protease inhibitor Kazal type 1 (SPINK1) gene are associated with various forms of human CP. We previously showed that deletion of Spink3, the mouse homologue of SPINK1, causes pancreatitis-like changes in the mouse. Spink3-/mice die within 2 weeks after birth, making it impossible to monitor long-term effects of Spink3 deficiency. The aim of this study was to rescue the Spink3-/phenotype by generating Spink3-/mice with knocked-in SPINK1, and to characterize timedependent changes in the pancreas in this geneticmodel. Methods:We placedCAG promoterSPINK1 minigene (CAG-SP1) into diaphanous homolog 2 (Diap2) locus, which is located on X chromosome, using the Cre-Lox technology. X-inactivation is a process whereby one of the two copies of the X chromosome present in female mammals is inactivated. By utilizing X-inactivation, we were able to create mice in which SPINK level was partially, but not completely, reduced compared with the wild type. The CAG-SP1 knock-in mice were crossed to Spink3+/mice, thus generating Spink3-/-; CAG-SP1 knock-in mice. Mice were followed up for up to 6 months. Time-dependent changes in pancreatic histology, autophagy, inflammatory infiltration, acinar cell death, stellate cell activation, fibrosis, as well as trypsinogen activation and serum amylase were measured. Results: Spink3-/-;YXCAG-SP1, as well as Spink3-/-;XCAG-SP1/CAG-SP1 mice in which CAG-SP1 was present on both X chromosomes, showed no abnormalities in pancreas or any other organ during the 6 months of observation, indicating that human Spink1 rescues Spink3 deficiency in mice. Spink3-/-; XCAG-SP1/wild mice in which CAG-SP1 is present on only one of the two X chromosomes, showed slight growth retardation but were healthy and fertile. Pancreas of Spink3-/-; XCAGSP1/wildmice at birth contained both normal and degenerated acinar cells, with accumulation of autophagic vacuoles. The Spink3-/-;XCAG-SP1/wild mice time-dependently developed pathologic features of CP, including loss of acinar cells, intralobular fibrosis with activated stellate cells, and neutrophilic infiltration. In the interlobular areas, fibrosis was not evident, but instead prominent lipomatosis was observed. Interlobular ducts were dilated and contained protein plugs, which resembled CP in human. Older mice displayed acinar-ductal metaplasia and prominent expression of proto-oncogenes Egfr, Her2, and Ras, but did not develop pancreatic intraepithelial neoplasia. Conclusions: The results re-inforce the role of SPINK1/Spink3 deficiency in the development of CP and indicate that CP conditions trigger factors promoting pancreatic cancer development.

Su1239 A Prospective Trial of Secretin Infusion for Refractory Type B Pain in Chronic Pancreatitis

Background & Aims: Chronic pancreatitis (CP) is characterized by chronic inflammation and progressive pancreatic fibrosis leading eventually to the loss of exocrine and endocrine pancreas functions. The loss-of-function mutations of serine protease inhibitor Kazal type 1 (SPINK1) gene are associated with various forms of human CP. We previously showed that deletion of Spink3, the mouse homologue of SPINK1, causes pancreatitis-like changes in the mouse. Spink3-/mice die within 2 weeks after birth, making it impossible to monitor long-term effects of Spink3 deficiency. The aim of this study was to rescue the Spink3-/phenotype by generating Spink3-/mice with knocked-in SPINK1, and to characterize timedependent changes in the pancreas in this geneticmodel. Methods:We placedCAG promoterSPINK1 minigene (CAG-SP1) into diaphanous homolog 2 (Diap2) locus, which is located on X chromosome, using the Cre-Lox technology. X-inactivation is a process whereby one of the two copies of the X chromosome present in female mammals is inactivated. By utilizing X-inactivation, we were able to create mice in which SPINK level was partially, but not completely, reduced compared with the wild type. The CAG-SP1 knock-in mice were crossed to Spink3+/mice, thus generating Spink3-/-; CAG-SP1 knock-in mice. Mice were followed up for up to 6 months. Time-dependent changes in pancreatic histology, autophagy, inflammatory infiltration, acinar cell death, stellate cell activation, fibrosis, as well as trypsinogen activation and serum amylase were measured. Results: Spink3-/-;YXCAG-SP1, as well as Spink3-/-;XCAG-SP1/CAG-SP1 mice in which CAG-SP1 was present on both X chromosomes, showed no abnormalities in pancreas or any other organ during the 6 months of observation, indicating that human Spink1 rescues Spink3 deficiency in mice. Spink3-/-; XCAG-SP1/wild mice in which CAG-SP1 is present on only one of the two X chromosomes, showed slight growth retardation but were healthy and fertile. Pancreas of Spink3-/-; XCAGSP1/wildmice at birth contained both normal and degenerated acinar cells, with accumulation of autophagic vacuoles. The Spink3-/-;XCAG-SP1/wild mice time-dependently developed pathologic features of CP, including loss of acinar cells, intralobular fibrosis with activated stellate cells, and neutrophilic infiltration. In the interlobular areas, fibrosis was not evident, but instead prominent lipomatosis was observed. Interlobular ducts were dilated and contained protein plugs, which resembled CP in human. Older mice displayed acinar-ductal metaplasia and prominent expression of proto-oncogenes Egfr, Her2, and Ras, but did not develop pancreatic intraepithelial neoplasia. Conclusions: The results re-inforce the role of SPINK1/Spink3 deficiency in the development of CP and indicate that CP conditions trigger factors promoting pancreatic cancer development.

Is Embolization of the Pancreas Safe? Pancreatic Histological Changes after Selective Transcatheter Arterial Embolization with N-Butyl Cyanoacrylate in a Swine Model

This study was designed to evaluate the safety of selective transcatheter arterial embolization (TAE) with N-butyl cyanoacrylate (NBCA) in a swine model in terms of histological changes in the pancreas. Three groups of two female swine (58-64kg) per group underwent TAE of the dorsal pancreatic artery, under anesthesia, with 1:1, 1:4, and 1:9 mixtures of NBCA and iodized oil. Blood parameters were evaluated at days 1, 4, and 10 after TAE, after which the animals were sacrificed and pancreatic tissues were examined under light microscopy. All of the animals were asymptomatic and survived for 10days. Cone beam computed tomographic angiography revealed occlusion of the dorsal pancreatic artery and no enhancement in the embolized area. The white blood cell count and C-reactive protein level were elevated slightly on day 1 after TAE (mean±SD: 252.7±27.8×10(2)/μl and 0.15±0.07mg/l, respectively), but they normalized or remained near the upper normal limit thereafter. The serum amylase and lipase levels also were elevated on day 1 (8831.7±2169.2 U/l and 130±53.4 U/l, respectively) but normalized thereafter. Histologically, necrosis and fibrosis were noted only in the embolized segment, and necrosis and acute inflammatory reactions were absent in the nonembolized segment. The border between both segments was well defined. Lymphocytic infiltration and foreign body reaction were noted around the embolized vessels. Selective TAE with NBCA in the pancreas caused localized ischemic necrosis without clinically significant pancreatitis; therefore, this procedure is tolerable in swine.