Agnieszka Madro

Can we expect progress in the treatment of fibrosis in the course of chronic pancreatitis

Chronic pancreatitis (CP) is a necroinflammatory process characterized by loss of both exocrine and endocrine function. To date, the disease has been treated symptomatically. Real advances in CP management can be expected once the pathophysiology of the disease is elucidated and individual stages of its development are properly managed. A key role in the CP pathogenesis is played by activation of pancreatic stellate cells (PSCs) that cooperate with the remaining pancreatic cells. All these cells produce cytokines, growth factors, angiotensin and other substances, which paracrinally or autocrinally induce further, persistent activation of PSCs. The activated PSCs are capable of producing and modifying the extracellular matrix. An optimal therapeutic preparation should exert beneficial effects on all the above-mentioned phenomena observed in CP. The most promising treatment modalities include blocking of the renin-angiotensin system (RAS), activation of peroxisome proliferator-activated receptors gamma (PPAR-γ), influence on the remaining PSC signaling pathways, blocking of substances produced by activated PSCs, and antioxidants. The findings of many recent experimental studies are highly encouraging; however, their efficacy should be confirmed in well-designed clinical trials.

Activated and inactivated PPARs-gamma modulate experimentally induced colitis in rats

Summary Background This study sought to define the mechanism by which PPAR-γ ligands affect the course of experimentally induced colitis in rats. Material/Methods Inflammation was induced in Wistar rats by a single rectal administration of 2,4,6,-trinitrobenzene sulfonic acid (TNBS). The antagonist of PPARγ antagonist, bisphenol A diglycidyl ether (BADGE), was administrated intraperitoneally 120 mg/kg 4 times every other day. Rosiglitazone 8 mg/kg was administrated by gastric tube 4 times. Body weight was measured daily. After killing, the large intestinal tissue was weighed and collected for histopathologic and immunoenzymatic tests. Levels of IL-6, IL-10, and myeloperoxidase (MPO) were determined in serum and in intestinal homogenates. Results Rats receiving rosiglitazone had higher body weight, whereas large intestine weight/length ratio was lower; histology showed fewer inflammatory markers. Rats receiving TNBS and TNBS along with BADGE had more intensive inflammatory changes. Rosiglitazone alone decreased expression of IL-6; used with TNBS it decreased expression of MPO in intestinal tissue, yet did not increase the expression of IL-10. Decreased levels of MPO indicate reduced neutrophil-dependent immune response. The antagonist of PPAR-γ increased IL-6 in serum and decreased IL-10 in intestinal homogenates. Bisphenol A diglycidyl ether administrated to healthy animals increases serum IL-6 levels. Conclusions Rosiglitazone inhibits experimental inflammation; administration of its selective antagonist abolishes this protective influence. Rosiglitazone inhibits expression of proinflammatory IL-6 and does not affect IL-10. Agonists of PPARs-γ are possibilities for inflammatory bowel disease prevention. Exogenous substances blocking PPARs-γ may contribute to development or relapse of nonspecific inflammatory bowel diseases.

Angiogenesis-related biomarkers in patients with alcoholic liver disease: their association with liver disease complications and outcome.

Angiogenesis is believed to be implicated in the pathogenesis of alcoholic liver disease (ALD). We aimed to explore the usefulness and accuracy of plasma angiogenic biomarkers for noninvasive evaluation of the severity of liver failure and ALD outcome. One hundred and forty-seven patients with ALD were prospectively enrolled and assessed based on their (1) gender, (2) age, (3) severity of liver dysfunction according to the Child-Turcotte-Pugh and MELD scores, and (4) the presence of ALD complications. Plasma levels of vascular endothelial growth factor (VEGF-A) and angiopoietins 1 and 2 (Ang1 and Ang2) were investigated using ELISAs. Multivariable logistic regression was applied in order to select independent predictors of advanced liver dysfunction and the disease complications. Significantly higher concentrations of Ang2 and VEGF-A in ALD patients as compared to controls were found. There was no difference in Ang1 levels in both groups. A positive correlation of Ang2 levels with INR (Rho 0.66; P < 0.0001) and its inverse correlation with plasma albumin levels (Rho –0.62; P < 0.0001) were found. High Ang2 concentrations turned out to be an independent predictor of severe liver dysfunction, as well as hepatic encephalopathy and renal impairment. Ang2 possessed the highest diagnostic and prognostic potential among three studied angiogenesis-related molecules.

The decrease of serum MMP-2 activity corresponds to alcoholic cirrhosis stage

Because of numerous limitations for liver biopsy, a noninvasive marker of liver cirrhosis is sought. Promising indicators seem to be matrix metalloproteinases (MMPs) that are responsible for degradation of extracellular matrix. The aim of the study was to evaluate the gelatinase activities (MMP-2 and MMP-9) in patients with different stages of alcoholic cirrhosis. Sixty-seven outpatients who presented various stages of alcoholic cirrhosis according to Child-Turcotte-Pugh criteria and 26 healthy control subjects were enrolled. Blood samples were collected for MMP-2 and MMP-9 activities. A significant decrease of serum MMP-2 activity was noted in stages B and C of cirrhosis in comparison with control. Serum MMP-9 activity did not depend on the stage of cirrhosis. The MMP-2 levels, but not those of MMP-9, may be of value in understanding the pathogenesis and progression of alcoholic cirrhosis.

The Genetic Predisposition and Its Impact on the Diabetes Mellitus Development in Patients with Alcoholic Chronic Pancreatitis

The most common cause of chronic pancreatitis (CP) is alcohol abuse. The aim of the present study was to identify patients with genetic predisposition to CP abusing alcohol. The question posed was whether CP manifests at a younger age and diabetes mellitus develops earlier in individuals with genetic predisposition. The study encompassed 79 patients with alcoholic chronic pancreatitis (ACP) and control group (100 persons). The following mutations were determined: R122H and N29I of PRSS1 and N34S of SPINK1 as well as E366K and E288V of SERPINA 1. No R122H and N291 mutations were observed in the group of ACP patients and in controls. Moreover, there was no E288V mutation. In 79 ACP patients, six SPINK 1 (N34S/wt) mutations were observed. In the control group, one heterozygous SPINK 1N34S gene mutation was found (P = 0.0238). Two PiZ mutations were identified in patients with ACP and one analogical mutation in controls. Amongst patients with ACP as well as SPINK1 and PiZ mutations, the onset of disease was observed earlier and developed earlier. The prevalence of SPINK1 mutation is higher in patients with ACP than in healthy populations. This mutation together with the effects of alcohol accelerates the development of ACP and of diabetes mellitus.

Tu1665 Role of Ghrelin and Melatonin in Pathogenesis of Clinical and Experimental Inflammatory Bowel Disease

model of tumourogenesis in SPARC knockout (KO) and wild-type (WT) mice. Methods: Colitis-associated tumourgenesis was promoted in SPARC-KO and WT mice by a single intraperitoneal injection of azoxymethane prior to three cycles of 7 days of dextran sodium sulphate. Tumour size, number and overall tumour burden was assessed endoscopically and histologically. Immunofluorescent (IF) assessment of CD4, CD8a, CD68, F4/80 and Ki-67 were performed on tumours. Tumours of similar size were harvested from both models at week 11 and processed for microsarray analysis. Whole-genome microarray (Illumina BeadChip, MouseRef 8 v2) compared gene expression in tumours from both genotypes. Results: All tumours were dysplastic tubular adenomas with focal high-grade dysplasia but SPARC-KO mice had a lower tumour burden than WT animals. IF assessment of the inflammatory cell infiltrates showed CD4-positive, CD8apositive, CD68-positive and F4/80-positive cells in all tumours without any significant differences between the models. KO tumours had higher numbers of KI-67+ve cells but this was not significant. Microarray analysis identified 6 differentially regulated genes between KO and WT mice (p < 0.05), 4 were decreased in KO tumours and were involved in the regulation of the immune response (Ifit2, Ido), cell growth and proliferation (Areg, Hoxa7). Two genes were increased in KO tumours with one associating with apoptosis (Erdr1) and one with an unknown function (Hamp2). Conclusions: Differences in gene expression were identified between SPARC-KO and WT tumours and these may impact on differences in tumour development and burden but not inflammatory cell infiltration.

The Influence of Serum MMP-2 Activity on the Development of Liver Fibrosis in Patients With Alcoholic Liver Cirrhosis

Background and Aims: Fatigue is a common symptom of chronic liver diseases (CLD), including non-alcoholic fatty liver disease (NAFLD) and chronic hepatitis C (CH-C). The mechanisms or correlates of fatigue in these patients have not been well studied. We aimed to determine if there is a correlation between self-reports of physical activity associated fatigue (peripheral fatigue) or more global lack of energy and motivation (central fatigue); with serum markers of inflammation, or with abnormalities of glucose and lipid metabolism. Methods: 31 untreated patients (age 52.5 ±6.8 years, 66.7% male, BMI 32.4 ± 5.5, 26.7% DM, 0% cirrhosis) with CLD (biopsy proven NAFLD or CH-C with viremia) participated in the study. Fasting blood samples were obtained and were assessed for levels of cytokines (IL-6, IL-8, and TNF-α), serotonin, C-peptide insulin, liver enzymes (AST, ALT), glucose, and lipids (triglycerides, total cholesterol, HDL, LDL, and the non-HDL fraction). Cytokines, serotonin, and C-peptide insulin were measured by ELISA following the manufacturers protocols. The remaining parameters were measured by the Cholestech LDX system. Selfreports included standardized and valid measures of depression (short form of CES-D), vitality/energy (vitality subscale of the SF36) and level of activity (Human Activity Profile). Patients were then divided into tertiles by MET value. The middle third was omitted from further analysis; the remaining top third (those with MET >8.8, representing strenuous activity) and bottom third (those with MET 7 and a transformed vitality index score <45. Only patients meeting these criteria were defined as having central fatigue. Groupwise comparisons were made by Mann-Whitney test, and correlations were assessed by Spearman Rho. Results: In comparison to CLD patients without peripheral fatigue, CLD patients who had peripheral fatigue (n=23) had significantly elevated serum levels of IL-6 (7.2±13.5 pg/mL vs. 1.6±0.74 pg/mL, p<0.01) and IL-8 (22.8±11.2 vs. 15.7±6.8 pg/mL, p<0.05); respectively. In terms of central fatigue, the ratio of AST/ALT was significantly lower in CLD patients with central fatigue than those CLD patients without central fatigue (0.862±0.166 vs. 1.118±0.283, p= 0.004). Conclusions: The current study demonstrates that a substantial majority of patients with CLD report significant peripheral fatigue. This type of fatigue is linked to elevated serum levels of IL-6 and IL-8, implying an inflammatory component present in patients with peripheral fatigue but not in those with central fatigue. Further study into the nature and extent of fatigue associated with CLD is warranted.