- Email: [email protected]
997 HESPERIDIN ATTENUATES LIPOPOLYSACCHARIDE-INDUCED FULMINANT HEPATIC FAILURE IN D-GALACTOSAMINE-SENSITIZED MICE
POSTERS phosphodiesterases were assessed in Sk-ChA-1 cells by Quantitative Real-Time PCR, western blot and immunohistochemistry. Results: Cyst fluid treatment of human cholangiocytes raises intracellular cAMP levels and cholangiocyte proliferation, which are significantly reduced by lanreotide treatment. We found that somatostatin receptor 5 is the most abundantly expressed somatostatin receptor in human cholangiocytes and induced by lanreotide treatment. There is a high basal expression of the cAMP metabolizing enzyme PDE4D in cholangiocytes which is significantly induced by lanreotide treatment. Blocking of SST5 with 8 nM BIM23056 or inhibition of PDE4D activity with 10 mM rolipram reverses the beneficial effect of lanreotide. Conclusions: Lanreotide significantly reduces PCLD patient cyst fluid-induced proliferation in a mechanism that depends on the somatostatin receptor 5 and the phosphodiesterase PDE4D. Therefore, reducing intracellular cAMP levels and cholangiocyte proliferation by the induction of PDE4D enzyme activity is an attractive novel target for the treatment of polycystic liver disease. 997 HESPERIDIN ATTENUATES LIPOPOLYSACCHARIDEINDUCED FULMINANT HEPATIC FAILURE IN D-GALACTOSAMINE-SENSITIZED MICE J.-Y. Wan1 , X. Gong2 , F.-L. Luo3 , H.-Z. Li4 , L. Zhang5 . 1 Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology, 2 Department of Anatomy, 3 Department of Pharmacy, The first Affiliated Hospital of Chongqing Medical University, 4 Department of Pharmacology, 5 Department of Pathophysiology, Chongqing Medical University, Chongqing, China E-mail: [email protected] Background and Aims: Fulminant hepatic failure (FHF) remains an extremely poor prognosis and high mortality; better treatments are urgently needed. Hesperidin, a flavanone glycoside isolated from Poncirus trifoliate, has been described to exhibit anti-inflammatory activities in several inflammatory models. However, the effects of Hesperidin on FHF are poorly understood. The present study was undertaken to investigate whether hesperidin is efficacious against Lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced FHF in mice and its potential mechanisms. Methods: Hesperidin (30, 100 and 150 mg/kg/d) was pretreated orally once daily for 3 days before LPS/D-GalN injected in mice. The mortality, hepatic tissue histology, plasma levels of Tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT) and aspartate aminotransferase (AST), hepatic tissue TNFalpha, myeloperoxidas (MPO) activity, caspase-3 activity and hepatocellular apoptosis were measured. Western blotting analysis of phosphor-inhibit kappa B (I-kappaB), phospho-p38 mitogenactivated protein kinase (phospho-p38 MAPK), phospho-c-jun N-terminal kinase (phospho-JNK) and phospho-extracellular signal regulated kinase (phospho-ERK) were determined. Besides, heme oxygenase-1 (HO-1) protein and activity in liver tissues was assayed. Results: Our data showed that administering hesperidin to mice reduced mortality and improved liver injury induced by LPS/D-GalN in a dose-dependent manner. In addition, TET dose-dependently inhibited LPS/D-GalN-induced NF-kappaB and MAPKs activation, TNF-alpha production, myeloperoxidas (MPO) activity, caspase-3 activation and hepatocellular apoptosis. Moreover, hesperidin dosedependently increased HO-1 protein expression and activity. Further, these protective effects of hesperidin against LPS/D-GalNinduced FHF were blocked by ZnPP IX, a HO-1 inhibitor. Conclusions: These results suggest that hesperidin has remarkable hepatoprotective effects on LPS/D-GalN-induced FHF and the possible mechanism is related to up-regulation of protein, which lead to inhibiting NF-kappaB and MAPKs, decreasing TNF-alpha production.
998 CENTROMERE PROTEIN A IS OVEREXPRESSED IN HEPATOCELLULAR CARCINOMA AND INVOLVED IN CELL CYCLE REGULATION Y. Li, Z. Zhu, S. Zhang, D. Yu, H. Yu, L. Liu, X. Cao, L. Wang, H. Gao, M. Zhu. Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai, China E-mail: [email protected] Background and Aims: Centromere protein A (CENP-A) plays important roles in cell cycle regulation and genetic stability. The association of CENP-A with hepatocarcinogenesis remains unknown. The aim of this study is to further confirm the status of CENP-A in liver cancer cells and to investigate the role of CENP-A in cell cycle regulation in hepatocarcinogenesis. Methods: Plasmids expressing the full length CENP-A or short interfering RNA targeting CENP-A were constructed and transfected into HepG2 cells. The biological activities, the expression of genes associated with cell cycle and apoptosis in these transfectants were analyzed by a series of assays. Additionally, the expression of CENP-A and clinicopathological features of hepatocellular carcinomas were analyzed. Results: CENP-A was overexpressed at mRNA and protein levels in hepatoma HepG2 cells and human hepatocellular carcinoma tissues. Small RNA interference targeting CENP-A partially inhibited HepG2 growth in vitro and in vivo by blocking cell cycle at G0-G1 phase through mediation of P21WAF1 and SKP-2, and by promoting apoptosis of HepG2 cells as indicated by the changes of MDM2 and Bcl-2/Bax. CENP-A overexpression was positively correlated with a high Ki-67 index, P53 protein, histological grade in human hepatocellular carcinomas. Conclusions: To the best of our knowledge, this is the first study showing overexpression of CENP-A in hepatoma cell lines and human HCC tissues. CENP-A might serve as a cell proliferation marker in cell cycle regulation and a potential therapeutic target for hepatocellular carcinoma. 999 HUMAN MONOCYTE SUBSETS ACCUMULATE INTRAHEPATICALLY IN LIVER CIRRHOSIS AND DIFFERENTIALLY INTERACT WITH HEPATIC STELLATE CELLS H.W. Zimmermann1 , S. Seidler1 , J. Nattermann2 , N. Gassler3 , C. Hellerbrand4 , R. Weiskirchen5 , C. Trautwein1 , F. Tacke1 . 1 Medizinische Klinik III, Universit¨ atsklinikum Aachen, Aachen, 2 Universit¨ at Bonn, Bonn, 3 Institut f¨ ur Pathologie, Universit¨ atsklinikum Aachen, Aachen, 4 Universit¨ at Regensburg, Regensburg, 5 Institut f¨ ur Klinische Chemie und Pathobiochemie, Universit¨ atsklinikum Aachen, Aachen, Germany E-mail: [email protected] Background and Aims: Monocyte-derived macrophages critically perpetuate inflammatory responses after liver injury as a prerequisite for organ fibrosis. Experimental murine models identified an essential role for the CCR2-dependent infiltration of classical Gr1/Ly6C+ monocytes in hepatic fibrosis. The relevance of monocyte subpopulations for human liver fibrosis is unknown. We therefore addressed the role of circulating and intrahepatic CD14++ CD16− and CD14+ CD16+ monocytes in patients with chronic liver diseases. Materials and Methods: Circulating blood monocyte subsets were studied by FACS in 226 chronic liver disease patients and 184 healthy controls. Liver specimens were analyzed by FACS and immunofluorescence for the presence and activation of monocyte/macrophage subsets. Moreover, isolated monocyte subsets were functionally characterized in vitro as well as cocultured with primary human hepatic stellate cells (HSC) to elucidate possible interactions between both cell types.
Journal of Hepatology 2010 vol. 52 | S319–S457
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998 CENTROMERE PROTEIN A IS OVEREXPRESSED IN HEPATOCELLULAR CARCINOMA AND INVOLVED IN CELL CYCLE REGULATION Y. Li, Z. Zhu, S. Zhang, D. Yu, H. Yu, L. Liu, X. Cao, L. Wang, H. Gao, M. Zhu. Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai, China E-mail: [email protected] Background and Aims: Centromere protein A (CENP-A) plays important roles in cell cycle regulation and genetic stability. The association of CENP-A with hepatocarcinogenesis remains unknown. The aim of this study is to further confirm the status of CENP-A in liver cancer cells and to investigate the role of CENP-A in cell cycle regulation in hepatocarcinogenesis. Methods: Plasmids expressing the full length CENP-A or short interfering RNA targeting CENP-A were constructed and transfected into HepG2 cells. The biological activities, the expression of genes associated with cell cycle and apoptosis in these transfectants were analyzed by a series of assays. Additionally, the expression of CENP-A and clinicopathological features of hepatocellular carcinomas were analyzed. Results: CENP-A was overexpressed at mRNA and protein levels in hepatoma HepG2 cells and human hepatocellular carcinoma tissues. Small RNA interference targeting CENP-A partially inhibited HepG2 growth in vitro and in vivo by blocking cell cycle at G0-G1 phase through mediation of P21WAF1 and SKP-2, and by promoting apoptosis of HepG2 cells as indicated by the changes of MDM2 and Bcl-2/Bax. CENP-A overexpression was positively correlated with a high Ki-67 index, P53 protein, histological grade in human hepatocellular carcinomas. Conclusions: To the best of our knowledge, this is the first study showing overexpression of CENP-A in hepatoma cell lines and human HCC tissues. CENP-A might serve as a cell proliferation marker in cell cycle regulation and a potential therapeutic target for hepatocellular carcinoma. 999 HUMAN MONOCYTE SUBSETS ACCUMULATE INTRAHEPATICALLY IN LIVER CIRRHOSIS AND DIFFERENTIALLY INTERACT WITH HEPATIC STELLATE CELLS H.W. Zimmermann1 , S. Seidler1 , J. Nattermann2 , N. Gassler3 , C. Hellerbrand4 , R. Weiskirchen5 , C. Trautwein1 , F. Tacke1 . 1 Medizinische Klinik III, Universit¨ atsklinikum Aachen, Aachen, 2 Universit¨ at Bonn, Bonn, 3 Institut f¨ ur Pathologie, Universit¨ atsklinikum Aachen, Aachen, 4 Universit¨ at Regensburg, Regensburg, 5 Institut f¨ ur Klinische Chemie und Pathobiochemie, Universit¨ atsklinikum Aachen, Aachen, Germany E-mail: [email protected] Background and Aims: Monocyte-derived macrophages critically perpetuate inflammatory responses after liver injury as a prerequisite for organ fibrosis. Experimental murine models identified an essential role for the CCR2-dependent infiltration of classical Gr1/Ly6C+ monocytes in hepatic fibrosis. The relevance of monocyte subpopulations for human liver fibrosis is unknown. We therefore addressed the role of circulating and intrahepatic CD14++ CD16− and CD14+ CD16+ monocytes in patients with chronic liver diseases. Materials and Methods: Circulating blood monocyte subsets were studied by FACS in 226 chronic liver disease patients and 184 healthy controls. Liver specimens were analyzed by FACS and immunofluorescence for the presence and activation of monocyte/macrophage subsets. Moreover, isolated monocyte subsets were functionally characterized in vitro as well as cocultured with primary human hepatic stellate cells (HSC) to elucidate possible interactions between both cell types.
Journal of Hepatology 2010 vol. 52 | S319–S457
S385