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Expression of p53,Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease

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World Journal of Gastroenterology 2006年第38期12卷 6198-6202页

作者： anatol panasiuk Janusz Dzieciol Bozena panasiuk Danuta Prokopowicz Department of Infectious Diseases Medical University of Biaystok Poland Department of Human Anatomy Medical University of Bialystok Poland

AIM: To analyze the protein expression essential for apoptosis in liver steatosis. METHODS: The expression of proapoptotic proteins p53, Bax, and antiapoptotic Bcl-2 in hepatocytes with steatosis (SH) and without steatosis (NSH) was evaluated in 84 patients at various stages of non-alcoholic fatty liver disease (NAFLD). RESULTS: Immunohistochemical staining of liver tissue showed the activation of p53 protein in SH and NSH with increased liver steatosis, diminished Bcl-2 and slightly decreased Bax protein. Positive correlation was found between the stage of liver steatosis with p53 expression in SH (r = 0.54, P < 0.01) and NSH (r = 0.49, P < 0.01). The antiapoptotic protein Bcl-2 was diminished together with the advancement of liver steatosis, especially in non-steatosed hepatocytes (r =0.43, P < 001). CONCLUSION: Apoptosis is one of the most important mechanisms leading to hepatocyte elimination in NAFLD. The intensification of inflammation in NAFLD induces proapoptotic protein p53 with the inhibition of antiapoptotic Bcl-2.

关键词： Apoptosis Non-alcoholic liver disease p53 Bcl-2 Bax Immunohistochemistry

Effect of interferon alpha2b plus ribavirin treatment on selected growth factors in respect to inflammation and fibrosis in chronic hepatitis C

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World Journal of Gastroenterology 2005年第12期11卷 1854-1858页

作者： panasiuk anatol Flisiak Robert Prokopowicz Danuta Department of Infectious Diseases

AIM: Growth factors (GF) that participate in regeneration and apoptosis have an important role in chronic liver diseases. We analyzed serum GF concentration during antiviral treatment and correlated it with morphological liver failure in chronic hepatitis C. METHODS: The levels of GF were determined in sera by ELISA method in 0,16,32 and 48 wk of therapy in 40 patients treated with IFNα2b (9 MU sc/wk) and RBV (1.2 g/d) and in 25 healthy subjects. Blind liver biopsies were done before treatment with histological grading and staging examination. RESULTS: The hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were markedly elevated prior the treatment and decreased during the therapy, although they did not reach the normal level. In non-responding (NR) patients, HGF and EGF were higher than that in responders (R), however differences were not significant. Before the treatment thrombopoietin (TPO) level was significantly lower in R than in NR (P<0.03). Platelet-derived growth factor (PDGF) concentration was lower in chronic hepatitis C than in healthy subjects and decreased during the treatment. A significant positive correlation was observed between inflammatory activity in the liver tissue and the concentration of HGF (in R: r= 0.4, in NR: r= 0.5), TPO (R: r= 0.6), and a significant negative correlation between this activity and EGF (R: r = -0.6) and PDGF (R: r= -0.5). Serum HGF concentration was higher in more advanced fibrosis (R: r = 0.5, P<0.05; NR: r=0.4, P<0,03). CONCLUSION: The decrease in PDGF can be an effective prognostic marker of the treatment and HCV elimination. Decreasing HGF, EGF, and PDGF can influence the inhibition of inflammatory and fibrotic processes in the liver during the antiviral treatment.

关键词： Growth factor Chronic hepatitis C Liver biopsy Interferon alfa 2b Ribivirin

Expression of β2-integrin on leukocytes in liver cirrhosis

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World Journal of Gastroenterology 2006年第38期12卷 6193-6197页

作者： anatol panasiuk Janusz ■ak Elzbieta Maciorkowska Bozena panasiuk Danuta Prokopowicz Department of Infectious Diseases Medical University of Bialystok Poland Department of Pediatrics Diagnostics Medical University of Bialystok Poland Department of Pediatric Nursing Medical University of Bialystok Poland

AIM： To analyze β2-integrin expression on blood leukocytes in liver drrhosis.METHODS： In 40 patients with liver cirrhosis and 20 healthy individuals, the evaluation of expression of CD11a （LFA-1α）, CD11b （Mac-1α）, CD11c （αX） and CD49d （VLA-4α） on peripheral blood leukocytes was performed using flow cytometry. The analysis was carried out in groups of patients divided into B and C according to Child-Pugh＇s classification. RESULTS： An increased CDlla, CD11b, CD11c and CD49d integrin expression was observed on peripheral blood leukocytes in liver cirrhosis. The integrin levels were elevated as the advancement of liver failure progressed. The highest expression of integrins occurred predominantly on monocytes. A slight expression of VLA-4 was found on lymphocytes and granulocytes and it increased together with liver failure. A positive correlation was noted between median intensity of fluorescence （MIF） expression on polymorphonuclear cells of CD11a and CD11c and CD49d （r = 0.42, P 〈 0.01; r = 053, P 〈 0.01, respectively） in liver cirrhosis stage C. However, no correlation was observed between integrin expression on leukocytes. The concentrations of sICAM-1, sVCAM-1, and TNFα, were significantly elevated in liver cirrhosis. CONCLUSION： β2-integrin expression on leukocytes increases in liver cirrhosis decornpensated as the stage of liver failure increases, which is a result of permanent activation of leukooytes circulating through the inflamed liver environment.β2-integrin expression on circulating leukocytes can intensify liver cirrhosis,

关键词： β2-integrin Liver cirrhosis Flow cytometry Leukocytes

Concentrations of gastric mucosal cytokines in children with food allergy and Helicobacterpylori infection

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World Journal of Gastroenterology 2005年第43期11卷 6751-6756页

作者： El(z|.)bieta Maciorkowska anatol panasiuk Maciej Kaczmarski Department of Pediatric Nursing Department of Infectious Diseases 3rd Department of Children's Diseases

AIM： To measure the concentrations of chosen cytokines in the antrum mucosa depending on the kind of harmful pathogenic factors and to compare the concentrations with the values of controls without allergy and coexisting Helicobacter pylori （H pylori） infection. METHODS： The patients （97 children） were divided into three groups according to the data obtained from the case history, to the main cause of the disease and to the dominant clinical symptoms. Group Ⅰ： children with food allergy （Fa）; group Ⅱ： children infected with H pylori; group Ⅲ （control group）： children with functional disorders of the alimentary tract （without Fa and Hp infection）. H pylori infection was determined by the presence of anti-Hp antibodies in serum （ELISA method） and urease test performed during endoscopic examination. Cytokine concentration in homogenates of gastric mucosa was detected by ELISA method. RESULTS： The IL-2 concentration in gastric mucosa bioptates was the highest in children with Hp infection （116.5±179.5 pg/mg of the protein） and Fa and Hp infection （98.1±101.0 pg/mg）, while decreased in children with Fa （44.8±50.3 pg/mg） and controls （45.7±23.5 pg/mg）. The lowest mean concentration of IFN-y was observed in children with Hp infection （18.9±16.4 pg/mg）, with Fa and Hp infection （25.5±27.7 pg/mg）, with Fa （40.6±39.7 pg/mg） and controls （49.9±33.4 pg/mg）. The highest IL-4 concentrations were observed in children with Hp infection （35.3±52.8 pg/mg） and in children with Fa and Hp infection （37.2±51.7 pg/mg）, while lower IL-4 concentration （23.64-35.8 pg/mg） was found in children with Fa compared to the controls （22.7±13.8 pg/mg）. The analysis of IL-4 concentrations in children with Hp infection regarding the intensity of gastritis showed the highest value （62.2±61.2 pg/mg） in mild and moderate gastritis. The concentrations of IL-5 in the gastric mucosa of children with or without Fa did not differ significantly and were comparable to the control group. The highest mean IL-8 val

关键词： Mucosal cytokines Food allergy Helicobacterpylori dehydrogenase complex

Signal transduction pathways in liver and the influence of hepatitis C virus infection on their activities

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World Journal of Gastroenterology 2009年第18期15卷 2184-2189页

作者： Magdalena M Dabrowska anatol panasiuk Robert Flisiak Department of Infectious Diseases and Hepatology Medical University of Bialystok14 Zurawia Str15-540 BialystokPoland

In liver,the most intensively studied transmembrane and intracellular signal transduction pathways are the Janus kinase signal transduction pathway,the mitogen-activated protein kinases signal transduction pathway,the transforming growth factor β signal transduction pathway,the tumor necrosis factor α signal transduction pathway and the recently discovered sphingolipid signal transduction *** of them are activated by many different cytokines and growth *** regulate specific cell mechanisms such as hepatocytes proliferation,growth,differentiation,adhesion,apoptosis,and synthesis and degradation of the extracellular *** replication cycle of hepatitis C virus(HCV) is intracellular and requires signal transduction to the nucleus to regulate transcription of its ***,HCV itself,by its structural and non-structural proteins,could influence the activity of the second signal ***,the inhibition of the transmembrane and intracellular signal transduction pathways could be a new therapeutic target in chronic hepatitis C treatment.

关键词： Liver Hepatitis C virus infection Signaltransduction pathway Proliferation Apoptosis

Phagocytic and oxidative burst activity of neutrophils in the end stage of liver cirrhosis

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World Journal of Gastroenterology 2005年第48期11卷 7661-7665页

作者： anatol panasiuk Jolanta Wysocka Elzbieta Maciorkowska Bozena panasiuk Danuta Prokopowicz Janusz Zak Karol Radomski Department of Infectious Diseases Department of Pediatrics Diagnostics Department of Pediatric Nursing

AIM：To evaluate the phagocytic activity and neutrophil oxidative burst in liver ***： In 45 patients with advanced postalcoholic liver cirrhosis （aged 45±14 years） and in 25 healthy volunteers （aged 38±5 years）, the percentage of phagocytizing cells after in vitro incubation with E. coli （Phagotest Kit）, phagocytic activity （mean intensity of fluorescence, MIF） and the percentage of neutrophil oxidative burst （Bursttest Kit）, and the level of free oxygen radical production （MIF of Rodamine 123） were analyzed by flow cytometry. The levels of soluble sICAM-1, sVCAM-1, sP-selectin, sE-selectin, sL-selectin, and TNF-α were determined in blood ***： The percentage of E. coli phagocytizing neutrophils in liver cirrhosis patients was comparable to that in healthy subjects. MIF of neutrophil - ingested E. coli was higher in patients with liver cirrhosis. The oxidative burst in E. coli phagocytizing neutrophils generated less amount of active oxygen compounds in liver cirrhosis pa- tients （MIF of R123：24.7±7.1 and 29.7±6.6 in healthy, P〈0.01）. Phorbol myristate acetate （PMA） - stimulated neutrophilsproduced less reactive oxidants in liver cirrhosis patients than in healthy subjects （MIF of R123： 42.7±14.6 vs 50.2±13.3, P〈0.01）. A negative correlation was observed between oxidative burst MIF of PMA-stimulated neutrophils and ALT and AST levels （r -0.35, P〈0.05; r-0.4, P〈0.03）. sVCAM-1, sICAM-1, sE-selectin concentrations correlated negatively with the oxygen free radical production （MIF of R123） in neutrophils after PMA stimulation in liver cirrhosis patients （r-0.45, P〈0.05, r-0.41, P〈0.05; r-0.39, P〈0.05, respectively）.CONCLUSION： Neutrophil metabolic activity diminishes together with the intensification of liver failure. The metabolic potential of phagocytizing neutrophils is significantly lower in liver cirrhosis patients, which can be one of the causes of immune mechanism damage. The evaluation of oxygen metabolism of E. coli-stimulated neutrophils

关键词： Neutrophil Phagocytosis Oxidative burst Liver cirrhosis Flow cytometry

Blood platelet and monocyte activations and relation to stages of liver cirrhosis

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World Journal of Gastroenterology 2005年第18期11卷 2754-2758页

作者： anatol panasiuk Janusz Zak Edwina Kasprzycka Katarzyna Janicka Danuta Prokopowicz Department of Infectious Diseases Medical University of Bialystok Poland Department of Pediatrics Diagnostics Medical University of Bialystok Poland

AIM: Blood platelets (pIt) and monocytes are the cells that play a crucial role in the pathogenesis of liver damage and liver cirrhosis (LC). In this paper, the analysis of mutual relationship between platelets and monocytes activation in LC was conducted. METHODS: Immunofluorescent flow cytometry was used to measure the percentage of activated platelet populations (CD62P, CD63), the percentage of plt-monocyte aggregates (pma) (CD41/CD45), and activated monocytes (CD11b, CD14, CD16) in the blood of 20 volunteers and 40 patients with LC. Platelet activation markers: sP-selectin, platelet factor 4 (PF4), beta-thromboglobulin (PTG) and monocyte chemotactic peptide-1 (MCP-1) were measured and compared in different stages of LC. RESULTS: Platelet activation with the increase in both βTG serum concentration and elevation of pIt population (CD62P and CD63 as well as MIF CD62P and CD63) is elevated as LC develops and thrombocytopenia rises. There is a positive correlation between medial intensity of fluorescence (MIF) CD62P and MIF CD63 in LC. We did not show any relationship between monocyte activation and pma level. SP-selectin concentration correlates positively with pIt count and pma, and negatively with stage of pIt activation and MIF CD62P and MIF CD63. There was no correlation between MCP-1 concentration and pIt, monocyte activation as well as pma level in LC. CD16 monocytes and MIF CD16 populations are significantly higher in the end stage of LC. A positive correlation occurs between the value of CDllb monocyte population and MIF CD14 and MIF CD16 on monocytes in LC. CONCLUSION: Platelet and monocyte activation plays an important role in LC. Platelet activation stage does not influence monocyte activation and production of pIt aggregates with monocytes in LC. With LC development, thrombocytopenia may be the result of pIt consumption in platelet-monocyte aggregates.

关键词： Blood platelet Monocyte Liver cirrhosis Flow cytometry

Soluble adhesion molecules ICAM-1,VCAM-1,P-selectin in children with Helicobacter pylori infection

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World Journal of Gastroenterology 2005年第43期11卷 6745-6750页

作者： Elzbieta Maciorkowska Maciej Kaczmarski anatol panasiuk Katarzyna Kondej-Muszynska Andrzej Kemonai Department of Pediatric Nursing 3rd Department of Children's Diseases Department of Infectious Diseases Department of Pathomorphology

AIM: To assess the sICAM-1, sVCAM-1, and sP-selectin levels in children withHelicobacter pylori(H pylori)infection and to evaluate their significance for the morphological changes found in gastric ***: The study included 106 children: 59children (55.7%) with chronic gastritis and positive IgG against H pylori, 29 children (27.3%) after previous H pylori infection without the bacterium colonization but with positive IgG against H pylori, and 18 children (17%) with functional disorders of the gastrointestinal system but with normal IgG against H pylori. Endoscopic and histopathological evaluation of gastric mucosa was performed based on the Sydney System *** evaluation of sP-selectin, sIC AM-1, sVCAM-1 levels in the sera of children was carried out using ELISA ***: The assessment of gastritis activity degrees indicated statistically significant values in the antrum and corpus (P＜0.001) of children examined. Serum sVCAM-1 levels were higher in group with gastritis due to H pylori infection than in group without infection and differed statistically (P＜0.05). Serum sVCAM-1 levels proved to be the highest among other adhesive molecules in infected children and decreased after eradication of H pylori. Serum sICAM-1 levels were similar in all examined groups. Serum sP-selectin levels were similar in children with and without H pylori ***: Assessment of adhesive molecules (sPselectin, sICAM-1, sVCAM-1) in the sera of children with active H pylori infection can show the participation of sVCAM-1 in the pathogenesis of gastric mucosal inflammation, sP-selectin and sICAM-1 concentrations in the sera of children with H pylori infection after eradication cannot reveal any significant differences as compared to healthy children.

关键词： sVCAM-1 sICAM-1 sP-selectin Helicobacter pylori

Specific ssDNA concentration in liver tissue as an index of apoptosis in hepatitis C virus-infected patients

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World Journal of Gastroenterology 2005年第39期11卷 6130-6133页

作者： Tadeusz Wojciech Lapiski anatol panasiuk Jerzy Jaroszewicz Oksana Kowalczuk Robert Flisiak Magdalena Rogalska Department of Infectious Diseases Medical University of Bialystok Poland Department of Infectious Diseases Medical University of BialystokPolandDepartment of Infectious Diseases Medical University of BialystokPolandInstitute of Molecular Biology Medical University of BialystokPolandDepartment of Infectious Diseases Medical University of BialystokPolandDepartment of Infectious Diseases Medical University of BialystokPoland

AIM. To evaluate bhe activity of apoptosis in liver tissue and explore its possible association with hepatic necroinflammarion and fibrosis as well as serum hepatitis C virus （HCV） load. METHODS： The studied population included 50 chronic hepatitis C patients （20 women and 30 men, aged 18-66 years）. HCV-RNA quantification was performed by two-step real-time quantitative RT-PCR mebhod using the TaqMan technology （reagents of Applera Corporation firm, USA）. The morphology of liver tissue was assessed descriptively and scored （necroinflammatory activity and fibrosis）. The early apoptosis activity in liver tissue was examined by ssDNA apoptosis ELISA kit, （Chemicon, Germany）. RESULTS： The correlation between apoptosis and fibrosis in liver tissue was observed. High intensification of apoptosis was proportional to the increase of fibrosis （ssDNA： 16.65×10^-5μg/g; 12.71×10^-5μg/g）, however, bhis difference was not statistically significant （P〉0.05）. Activity of apoptosis in the liver tissue, expressed by ssDNA concentration did not depend on hepatic necroinflammatory changes, HCVRNA viral load, ALT, and AST activity as well as prothrombin time and INR index. CONCLUSION： Fibrosis in the tissue is closely associated with early apoptosis in HCV-infected patients.

关键词： Apoptosis Hepatitis C virus ssDNA Fibrosis