Cellular changes post hepatectomy in cirrhotic vs normal rats

ACKNOWLEDGEMENTS I would like to express my sincere gratitude to my supervisor Dr. Theresa Tan and cosupervisor Associate Professor Leow Chon Kar for their invaluable supervision, support and encouragement throughout the course of my study. I would also thank Professor Sit Kim Ping for her concern for my study. My heartfelt thanks also go to Loh Chien Yuen, Qu Bin, Sherry Ngo, Lai Liqi, Low Teck Yew, Lee Mui Khin, other lab mates and staffs of the Department of Biochemistry for their technical assistance, helpful discussions and suggestions. I also wish to thank the head of the Department of Biochemistry for allowing me to carry out my research in the department and National University of Singapore for the award of the research scholarship. Last but not the least, I would thank my husband, son and parents for their understanding and support throughout my study. CELLULAR CHANGES POST HEPATECTOMY IN CIRRHOTIC VS NORMAL RATS TABLE OF CONTENTS CHAPTER INTRODUCTION 1.1 1.1.1 1.1.2 1.2 1.2.1 1.2.2 1.3 1.3.1 1.3.2 1.3.3 1.3.4 1.3.5 1.4 1.4.1 1.4.2 1.4.2.1 1.4.2.2 1.4.2.3 1.4.2.4 1.4.2.5 1.5 1.5.1 1.5.2 1.5.3 1.5.4 1.5.4.1 1.5.5 1.6 Liver Liver regeneration Partial hepatectomy Experimental model of liver cirrhosis Liver cirrhosis Thioacetamide induced cirrhosis in rat Mitochondria oxidative phosphorylation Complex I Complex II Complex III Cytochrome c oxidase ATP synthase Oxidative stress Reactive oxygen species Antioxidants Glutathione (GSH) Superoxide dismutase (SOD) Glutathione peroxidase Glutathione reductase Catalase Liver regeneration related proteins and genes Cell cycle Priming role of TNF-α and IL-6 in liver regeneration Role of HGF and TGF-α in liver regeneration Role of cyclins, cdks and cdk inhibitors in cell cycle Structure of cdk2, cdk2-cyclin A and cdk2-cyclin A-p27 p53 Objectives of this study 16 17 21 21 21 23 24 25 26 26 27 28 29 29 33 33 34 34 35 35 36 36 37 38 39 40 41 42 CHAPTER MATERIALS AND METHODS 2.1 2.2 2.3 2.4 2.5 Chemicals Induction of cirrhosis Partial hepatectomy Preparation of mitochondria Measurement of protein concentration 44 45 45 46 46 2.6 2.7 2.8 2.9 2.10 2.11 2.12 2.13 2.14 2.15 2.16 2.17 2.17.1 2.17.2 2.18 2.18.1 2.18.2 2.19 Determination of the respiratory control and ADP/O ratios Mitochondrial NADH-cytochrome c reductase activity Mitochondrial succinate-cytochrome c reductase activity Mitochondrial cytochrome c oxidase activity Mitochondrial ATPase activity Hepatic and mitochondrial glutathione determination Mitochondrial and cytosolic superoxide dismutase activity Mitochondrial and cytosolic glutathione peroxidase activity Mitochondrial and cytosolic glutathione reductase activity Measurement of malondialdehyde Western blotting ELISA TNF-α ELISA IL-6 ELISA mRNA analysis RNA isolation RT-PCR Statistical analysis 47 47 48 48 49 49 50 50 51 51 52 54 54 55 56 56 57 59 CHAPTER INDUCTION OF CIRRHOSIS 3.1 3.2 3.3 3.4 Introduction Thioacetamide induced liver cirrhosis Effects on mitochondrial function and GSH levels Conclusions 61 62 62 62 CHAPTER MITOCHONDRIA FUNCTION 4.1 4.2 4.3 4.3.1 4.3.2 4.3.3 4.3.4 4.3.5 4.4 Introduction Respiratory enzymes activities in normal and cirrhotic livers Changes in mitochondrial respiratory enzymes and ATPase following Hepatectomy SCCR activity NCCR activity CCO activity Respiratory control and P/O ratios Mitochondrial ATPase activity Conclusions 66 68 69 69 69 72 72 72 79 CHAPTER OXIDATIVE STRESS 5.1 5.2 5.3 Introduction Mitochondrial and cytosolic antioxidant capacity in normal and cirrhotic livers Changes in mitochondrial and cytosolic antioxidant capacity following partial Hepatectomy 80 82 84 5.3.1 5.3.2 5.3.3 5.3.4 5.3.5 5.3.6 5.4 Mitochondrial and hepatic GSH level Mitochondrial and cytosolic superoxide dismutase activity Mitochondrial and cytosolic glutathione peroxidase activity Mitochondrial and cytosolic glutathione reductase activity Mitochondrial and cytosolic catalase expression Mitochondrial lipid peroxidation Conclusions 84 84 89 89 89 96 96 CHAPTER EXPRESSION PATTERNS OF CYTOKINE, GROWTH FACTOR AND CELL CYCLE-RELATED GENES 6.1 6.2 6.2.1 6.2.2 6.3 6.3.1 6.3.2 6.4 6.4.1 6.4.2 6.4.3 6.4.4 6.5 6.5.1 6.5.2 6.6 6.6.1 6.6.2 6.7 6.8 Introduction Hepatic TNF-α and IL-6 TNF-α level IL-6 level Expression of growth factors HGF mRNA TGF-α mRNA Expression of cyclins Expression of cyclin D1 Expression of cyclin D3 Expression of cyclin E Expression of cyclin A Expression of cdks Expression of cdk4 Expression of cdk2 mRNA of cdk inhibitors p21 mRNA p27 mRNA p53 mRNA Conclusions 98 100 100 100 101 101 101 102 102 102 110 110 110 110 114 114 114 114 118 118 CHAPTER DISCUSSION 7.1 7.2 7.3 7.4 7.5 7.6 Respiratory enzymes activities in normal and cirrhotic livers Changes in mitochondrial respiratory enzymes and ATPase following hepatectomy Mitochondrial and cytosolic antioxidant capacity in normal and cirrhotic livers Changes in mitochondrial and cytosolic antioxidant capacity following partial hepatectomy Expression of cyclins, cdks and cdk inhibitors in normal and cirrhotic livers Changes in expression of cyclins, cdks and cdk inhibitors after PH 120 121 122 124 126 126 7.7 7.8 7.9 Expression of cytokines and growth factors in normal and cirrhotic livers Changes in expression of cytokines and growth factors following PH Conclusions 128 129 132 Abstract For many liver malignancies, major hepatectomy is the main course of therapy. Although a normal liver has a tremendous capacity for regeneration, liver hepatectomy in humans is usually carried out on a diseased liver and in such instances, liver regeneration takes place in a cirrhotic remnant. As the demand of energy increases during liver regeneration, the mitochondria must play an important role. Cytokines and growth factors also play important roles in liver regeneration. Hepatocytes need to be primed before they are competent to respond to growth factors and proliferate. Cytokines such as TNF-α and IL6 can prime the hepatocytes, making them go from quiescent phase to G1 phase. Following this, the growth factors HGF and TGF-α act as mitogens and the hepatocytes respond by progressing through the cell cycle resulting in cell division. Cyclins such as cyclin D1, D3, E and A and their respective cyclin-dependant kinase partners play important roles at different phases of the cell cycle and regulate progress through the cell cycle. The activities of cdks can be inhibited by cdk inhibitors such as p21 and p27. This study aims to investigate how mitochondrial respiratory function, antioxidant capacity, cytokines, growth factors, cyclins, cdks and cdk inhibitors expressions change following partial hepatectomy of cirrhotic livers. Cirrhosis was induced in male WistarFurth rats by administration of thioacetamide. NADH-cytochrome c reductase activity, glutathione peroxidase activity and mitochondrial GSH levels were all significantly lowered in cirrhotic livers and in the cirrhotic remnants up to 72 h after 70% hepatectomy when compared to the corresponding controls. Lower respiratory control ratios with succinate as substrate were also observed at h till 48 h post-hepatectomy. At 24 h postPH, higher levels of lipid peroxidation were observed. Cirrhotic rats had decreased cytosolic IL-6 level till 72 h post-hepatectomy. Cirrhotic rats also had lower expressions of cyclin D1 from 24 h to 72 h; cyclin D3 at 24 h; cdk4 from h to 72 h; cylcin E till 72 h; cyclin A and cdk from 24 h to 48 h post-hepatectomy. But the mRNA levels of p21 at h and p27 from h to 48 h were up regulated in cirrhotic rats. In conclusion, when compared to the controls, cirrhotic livers have diminished oxidative phosphorylation capabilities due to changes in NADH-linked and FADH2-linked respiration as well as impaired antioxidant defenses following partial hepatectomy. Cirrhotic livers also have diminished cytokines, cyclins, cdks expression but increased cdk inhibitors expression. All these factors, if critical, could then impede liver regeneration. Keywords: partial hepatectomy, cirrhosis, mitochondria, cytokines, growth factors, cell cycle. Abbreviations ADP Adenosine 5’-diphosphate AP-1 Activated protein ATP Adenosine 5’-triphosphate CAK Cyclin-dependent kinase activation kinase CCO Cytochrome c oxidase cdk cyclin-dependent kinase Cip Cdk interacting protein Cyto Cytosolic ECM Extracellular matrix EDTA Ethylenediaminetetraacetic acid EGF Epidermal growth factor ELISA Enzyme-linked immunosorbent assay GPx Glutathione peroxidase GRd Glutathione reductase GSH Glutathione GSSG Glutathione disulfide GST Glutathione S-transferases HGF Hepatocyte growth factor HRP Horseradish peroxidase HSC Hepatic stellate cells Kip Kinase inhibitory protein IgG Immunoglobulin G IL-6 Interleukin-6 Ink4 Inhibitor of cdk4 MCB Monochlorobimane MDA Malondialdehyde Mt Mitochondrial NADH Reduced nicotinamide adenine dinucleotide NCCR NADH-cytochrome c reductase NF-κB Nuclear factor kappa B PCNA Proliferating cell nuclear antigen PH Partial hepatectomy pRb Retinoblastoma protein RCR Respiratory control ratio ROS Reactive oxygen species RT-PCR Reverse transcription-polymerase chain reaction SCCR Succinate-cytochrome c reductase SDS Sodium dodecyl sulfate SOD Superoxide dismutase STAT3 Signal transducers and activators of transcription TBA Thiobarbituric acid TBARS Thiobarbituric acid-reacting substances TBS-T Tris buffered saline-tween TEMED N, N, N’, N’-tetra-methylethylenediamine TGF Transforming growth factor TMB Tetramethyl-benzidine TNF-α Tumor necrosis factor-α TNFR Tumor necrosis factor-α receptor 10 References Abrahams JP, Leslie AG, Lutter R, Walker JE. Structure at 2.8 A resolution of F1ATPase from bovine heart mitochondria. Nature 1994; 370: 621-8. Ackrell BA. Progress in understanding structure-function relationships in respiratory chain complex II. FEBS Lett 2000; 466 (1): 1-5. Aguilar-Delfin I, Lopez-Barrera F, Hernandez-Munoz R. Selective enhancement of lipid peroxidation in plasma membrane in two experimental models of liver regeneration: partial hepatectomy and acute CCl4 administration. Hepatology 1996; 24: 657-62. Akerman P, Cote P, Yang SQ, McClain C, Nelson S, Bagby GJ, Diehl AM. Antibodies to tumor necrosis factor-alpha inhibit liver regeneration after partial hepatectomy. Am J Physiol 1992; 263: G579-85. Al-Bader A, Abul H, Hussain T, Al-Moosawi M, Mathew T, Dashti H. Selenium and liver cirrhosis. Mol Cell Biochem 1998; 185: 1-6. Albrecht JH, Rieland BM, Nelsen CJ, Ahonen CL. Regulation of G(1) cyclin-dependent kinases in the liver: role of nuclear localization and p27 sequestration. Am J Physiol 1999; 277: G1207-16. Albrecht JH, Poon RY, Ahonen CL, Rieland BM, Deng C, Crary GS. Involvement of p21 and p27 in the regulation of CDK activity and cell cycle progression in the regenerating liver. Oncogene 1998; 16: 2141-50. Albrecht JH, Meyer AH, Hu MY. Regulation of cyclin-dependent kinase inhibitor p21(WAF1/Cip1/Sdi1) gene expression in hepatic regeneration. Hepatology 1997; 25: 557-63. Albrecht JH, Hoffman JS, Kren BT, Steer CJ. Cyclin and cyclin-dependent kinase mRNA expression in models of regenerating liver and human liver diseases. Am J Physiol 1993; 265: G857-64. Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A 1993; 90: 7915-22. Arthur MJ. Collagenases and liver fibrosis. J Hepatol 1995; 22: 43-8. Baker SJ, Reddy EP. Transducers of life and death: TNF receptor superfamily and associated proteins. Oncogene 1996; 12: 1-9. 134 Balkan J, Dogru-Abbasoglu S, Kanbagli O, Cevikbas U, Aykac-Toker G, Uysal M. Taurine has a protective effect against thioacetamide-induced liver cirrhosis by decreasing oxidative stress. Hum Expt Toxicol 2001; 20: 251-4. Biagini G, Ballardini G. Liver fibrosis and extracellular matrix. J Hepatol 1989; 8: 11524. Bissell DM. Connective tissue metabolism and hepatic fibrosis: an overview. Semin Liver Dis 1990; 10: iii-iv. Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, Wolfson MF, Castner BJ, Stocking KL, Reddy P, Srinivasan S, Nelson N, Boiani N, Schooley KA, Gerhart M, Davis R, Fitzner JN, Johnson RS, Paxton RJ, March CJ, Cerretti DP. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature 1997; 385: 72933. Blindenbacher A, Wang X, Langer I, Savino R, Terracciano L, Heim MH. Interleukin is important for survival after partial hepatectomy in mice. Hepatology 2003; 38: 674-82. Boffoli D, Scacco S, Vergari R, Solarino G, Santacroce G, Papa S. (1994) Decline with age of respiratory chain activity in human skeletal muscle. Biochim Biophys Acta 1994; 1226: 73-82. Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med 1994; 331: 1286-92. Borowiak M, Garratt AN, Wustefeld T, Strehle M, Trautwein C, Birchmeier C. Met provides essential signals for liver regeneration. Proc Natl Acad Sci U S A 2004; 101: 10608-13. Bottenus R, Spach P, Filus S, Cunningham C. Effect of chronic ethanol consumption on energy-linked processes associated with oxidative phosphorylation: proton translocation and ATP-Pi exchange. Biochem Biophys Res Commun 1982; 105: 1368-73. Boyer PD. The ATP synthase--a splendid molecular machine. Annu Rev Biochem 1997; 66: 717-49. Brandt U. Proton-translocation by membrane-bound NADH:ubiquinone-oxidoreductase (complex I) through redox-gated ligand conduction. Biochim Biophys Acta. 1997; 1318 (1-2): 79-91. Broome U, Eriksson LS, Sundin U, Sundqvist KG. Decreased in vitro production of tumor necrosis factor in primary biliary cirrhosis patients. Scand J Gastroenterol 1992; 27: 124-8. 135 Cabre M, Camps J, Paternain JL, Ferre N, Joven J. Time-course of changes in hepatic lipid peroxidation and glutathione metabolism in rats with carbon tetrachloride-induced cirrhosis. Clin Exp Pharmacol Physiol 2000; 27: 694-9. Cadenas E. Biochemistry of oxygen toxicity. Annu Rev Biochem 1989; 58: 79-110. Carpenter G, Cohen S. Epidermal growth factor. J Biol Chem 1990; 265: 7709-12. Cederbaum A, Lieber C, Rubin E. Effects of chronic ethanol treatment on mitochondrial functions damage to coupling site I. Arch. Biochem Biophys 1974; 165: 560-9. Cerutti PA. Prooxidant states and tumor promotion. Science 1985; 227: 375-81. Chan SI, Li PM. Cytochrome c oxidase: understanding nature's design of a proton pump. Biochemistry 1990; 29 (1): 1-12. Cheeseman KH. Mechanisms and effects of lipid peroxidation. Mol Aspects Med 1993; 14 (3): 191-7. Chen J, Kunos G, Gao B. Ethanol rapidly inhibits IL-6-activated STAT3 and C/EBP mRNA expression in freshly isolated rat hepatocytes. FEBS Lett 1999; 457: 162-8. Chen Z, Putt DA, Lash LH. Enrichment and functional reconstitution of glutathione transport activity from rabbit kidney mitochondria: further evidence for the role of the dicarboxylate and 2-oxoglutarate carriers in mitochondrial glutathione transport. Arch Biochem Biophys 2000; 373: 193-202. Chen Z, Lash LH. Evidence for mitochondrial uptake of glutathione by dicarboxylate and 2-oxoglutarate carriers. J Pharmacol Exp Ther 1998; 285: 608-18. Chijiiwa K, Nakano K, Kameoka N, Nagai E Tanaka M. Proliferating cell nuclear antigen, plasma fibronectin, and liver regeneration rate after seventy percent hepatectomy in normal and cirrhotic rat. Surgery 1994; 116: 544-9. Cressman DE, Greenbaum LE, DeAngelis RA, Ciliberto G, Furth EE, Poli V, Taub R. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice. Science 1996; 274: 1379-83. Cressman DE, Diamond RH, Taub R. Rapid activation of the Stat3 transcription complex in liver regeneration. Hepatology 1995; 21: 1443-9. Cummings BS, Angeles R, McCauley RB, Lash LH. Role of voltage-dependent anion channels in glutathione transport into yeast mitochondria. Biochem Biophys Res Commun 2000; 276: 940-4. 136 Dashti H, Jeppsson B, Hagerstrand I, Hultberg B, Srinivas U, Abdulla M, Bengmark S. Thioacetamide- and carbon tetrachloride-induced liver cirrhosis. Eur Surg Res 1989; 21: 83-91. DeLeve LD, Kaplowitz N. Importance and regulation of hepatic glutathione. Semin Liver Dis 1990; 10: 251-66. Decker KF, Obolenskaya MY. Cytokines, nitric oxide synthesis and liver regeneration. J Gastroenterol Hepatol 1995; 10 Suppl 1: S12-7. Diehl AM, Yang SQ. Regenerative changes in C/EBP alpha and C/EBP beta expression modulate binding to the C/EBP site in the c-fos promoter. Hepatology 1994; 19: 447-56. Diehl AM, Yin M, Fleckenstein J, Yang SQ, Lin HZ, Brenner DA, Westwick J, Bagby G, Nelson S. Tumor necrosis factor-alpha induces c-jun during the regenerative response to liver injury. Am J Physiol 1994; 267: G552-61. Dyroff MC, Neal RA. Identification of the major protein adduct formed in rat liver after thioacetamide administration. Cancer Res 1981; 41: 3430-5. Edwards DR, Murphy G, Reynolds JJ, Whitham SE, Docherty AJ, Angel P, Heath JK. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J 1987; 6: 1899-904. Ehrenfried JA, Ko TC, Thompson EA, Evers BM. Cell cycle-mediated regulation of hepatic regeneration. Surgery 1997; 122: 927-35. el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B. WAF1, a potential mediator of p53 tumor suppression. Cell 1993; 75: 817-25. Estabrook R. Mitochondrial respiratory control and the polarographic measurement of ADP:O ratios. Methods Enzymol 1967; 10: 41-7. Esworthy RS, Ho YS, Chu FF. The Gpx1 gene encodes mitochondrial glutathione peroxidase in the mouse liver. Arch Biochem Biophys 1997; 340 (1): 59-63. Fausto N. Liver regeneration. J Hepatol 2000; 32: 19-31. Fausto N, Laird AD, Webber EM. Role of growth factors and cytokines in hepatic regeneration. FASEB J 1995; 9: 1527-36. Fernandez-Checa J, Kaplowitz N, Garcia-Ruiz C, Colell A, Miranda M, Mari M., Ardite E, Morales A. GSH transport in mitochondria: defense against TNF-induced oxidative stress and alcohol-induced defect. Gastrointest Liver Physiol 1997; 36: G7-17. 137 Fernandez-Checa J, Kaplowitz N. The use of monochlorobimane to determine hepatic GSH levels and synthesis. Anal Biochem 1990; 190: 212-9. FitzGerald MJ, Webber EM, Donovan JR, Fausto N. Rapid DNA binding by nuclear factor kappa B in hepatocytes at the start of liver regeneration. Cell Growth Differ 1995; 6: 417-27. Flohe L, Otting F. Superoxide dismutase assays. Methods Enzymol 1984; 105: 93-104. Flohe L, Gunzler W. Assays of glutathione peroxidase. Methods Enzymol 1984; 105: 114-21. Friedrich T, Steinmuller K, Weiss H. The proton-pumping respiratory complex I of bacteria and mitochondria and its homologue in chloroplasts. FEBS Lett 1995; 367 (2): 107-11. Franco D, Capussotti L, Smajda C, Brouzari H, Meakins J, Kemeny F, Grange D, Dellepiane M. Resection of hepatocellular carcinomas. Results in 72 European patients with cirrhosis. Gastroenterology 1990; 98: 733-8. Ganong W. Review of medical physiology. 20th edition. 2001. Garcia-Ruiz C, Morales A, Colell A, Rodes J, Yi JR, Kaplowitz N, Fernandez-Checa JC. Evidence that the rat hepatic mitochondrial carrier is distinct from the sinusoidal and canalicular transporters for reduced glutathione. Expression studies in Xenopus laevis oocytes. J Biol Chem 1995; 270: 15946-9. Gennis RB. How does cytochrome oxidase pump protons? Proc Natl Acad Sci U S A 1998; 95 (22): 12747-9. Girard F, Strausfeld U, Fernandez A, Lamb NJ. Cyclin A is required for the onset of DNA replication in mammalian fibroblasts. Cell 1991; 67: 1169-79. Gorina S, Pavletich NP. Structure of the p53 tumor suppressor bound to the ankyrin and SH3 domains of 53BP2. Science 1996; 274: 1001-5. Grana X, Reddy PE. Cell cycle control in mammalian cells: role of cyclins, cyclindependent kinases (CDKs), growth suppressor genes and cyclin dependent kinase inhibitors (CKIs). Oncogene 1995; 11: 211-9. Greenbaum LE, Li W, Cressman DE, Peng Y, Ciliberto G, Poli V, Taub R. CCAAT enhancer- binding protein beta is required for normal hepatocyte proliferation in mice after partial hepatectomy. J Clin Invest 1998; 102: 996-1007. 138 Griffith OW. Biologic and pharmacologic regulation of mammalian glutathione synthesis. Free Radic Biol Med 1999; 27: 922-35. Griffith OW, Meister A. Origin and turnover of mitochondrial glutathione. Proc Natl Acad Sci U S A 1985; 82: 4668-72. Guerrieri F, Vendemiale G, Grattagliano I, Cocco T, Pellecchia G, Altomare E. (1999) Mitochondrial oxidative alterations following partial hepatectomy. Free Rad Biol Med 1999; 26: 34-41. Guerrieri F, Muolo L, Cocco T, Capozza G, Turturro N, Cantatore P, Papa S. Correlation between rat liver regeneration and mitochondrial energy metabolism. Biochim Biophys Acta 1995; 1272(2): 95-100. Hagerhall C. Succinate: quinone oxidoreductases. Variations on a conserved theme. Biochim Biophys Acta 1997; 1320 (2): 107-41. Harada K, Shiota G, Kawasaki H. Transforming growth factor-alpha and epidermal growth factor receptor in chronic liver disease and hepatocellular carcinoma. Liver 1999; 19: 318-25. Hatefi Y. The mitochondrial electron transport and oxidative phosphorylation system. Annu Rev Biochem 1985; 54: 1015-69. Haupt Y, Maya R, Kazaz A, Oren M. Mdm2 promotes the rapid degradation of p53. Nature 1997; 387: 296-9. Hermolin J, Dmitriev OY, Zhang Y, Fillingame RH. Defining the domain of binding of F1 subunit epsilon with the polar loop of F0 subunit c in the Escherichia coli ATP synthase. J Biol Chem 1999; 274: 17011-6. Hernandez-Munoz R, Sanchez-Sevilla L, Marinez-Gomez A, Dent M. Changes in mitochondrial adenine nucleotides and in permeability transition in two models of rat liver regeneration. Hepatology 2003; 37: 842-51. Hernandez-Munoz R, Diaz-Munoz M, Lopez V, Lopez-Barrera F, Yanez L, Vidrio S, Aranda-Fraustro A, de Sanchez V. Balance between oxidative damage and proliferative potential in an experimental rat model of CCl4-induced cirrhosis: protective role of adenosine administration. Hepatology 1997; 26: 1100-10. Herwig S, Strauss M. The retinoblastoma protein: a master regulator of cell cycle, differentiation and apoptosis. Eur J Biochem 1997; 246: 581-601. Higami Y, Shimokawa I, Ando K, Tanaka K, Tsuchiya T. Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development. Cell Tissue Res 2000; 299: 363-9. 139 Higgins G, Anderson R. Experimental pathology of the liver: restoration of the white rat following partial surgical removal. Arch. Pathol 1931; 12: 186-202. Hirano T. Interleukin and its receptor: ten years later. Int Rev Immunol 1998; 16: 24984. Hortelano S, Dewez B, Genaro A, Diaz-Guerra M, Bosca L. (1995) Nitric oxide is released in regenerating liver after partial hepatectomy. Hepatology 1995; 21: 776-86. Huang ZZ, Chen C, Zeng Z, Yang H, Oh J, Chen L, Lu SC. Mechanism and significance of increased glutathione level in human hepatocellular carcinoma and liver regeneration. FASEB J 2001; 15: 19-21. Huang ZZ, Li H, Cai J, Kuhlenkamp J, Kaplowitz N, Lu SC. Changes in glutathione homeostasis during liver regeneration in the rat. Hepatology 1998; 27: 147-53. Hunter AL, Holscher MA, Neal RA. Thioacetamide-induced hepatic necrosis. I. Involvement of the mixed-function oxidase enzyme system. J Pharmacol Exp Ther 1977; 200: 439-48. Hwang C, Sinskey AJ, Lodish HF. Oxidized redox state of glutathione in the endoplasmic reticulum. Science 1992; 257: 1496-502. Inomoto T, Tanaka A, Mori S, Jin MB, Sato B, Yanabu N, Tokuka A, Kitai T, Ozawa K, Yamaoka Y. Changes in the distribution of the control of the mitochondrial oxidative phosphorylation in regenerating rabbit liver. Biochim Biophys Acta 1994; 1188: 311-7. Inoue Y, Tomiya T, Yanase M, Arai M, Ikeda H, Tejima K, Ogata I, Kimura S, Omata M, Fujiwara K. p53 May positively regulate hepatocyte proliferation in rats. Hepatology 2002; 36: 336-44. Iredale JP, Benyon RC, Arthur MJ, Ferris WF, Alcolado R, Winwood PJ, Clark N, Murphy G. Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. Hepatology 1996; 24: 176-84. Jaumot M, Estanyol JM, Serratosa J, Agell N, Bachs O. Activation of cdk4 and cdk2 during rat liver regeneration is associated with intranuclear rearrangements of cyclin-cdk complexes. Hepatology 1999; 29: 385-95. Jeffrey PD, Gorina S, Pavletich NP. Crystal structure of the tetramerization domain of the p53 tumor suppressor at 1.7 angstroms. Science 1995; 267: 1498-502. Johnson D, Lardy H. Isolation of liver or kidney mitochondria. Methods Enzymol 1967; 10: 94-6. 140 Jones PC, Fillingame RH. Genetic fusions of subunit c in the F0 sector of H+transporting ATP synthase. Functional dimers and trimers and determination of stoichiometry by cross-linking analysis. J Biol Chem 1998; 273: 29701-5. Kamiyama Y, Ozawa K, Honjo I. Changes in mitochondrial phosphorylative activity and adenylate energy charge of regenerating rabbit liver. J Biochem (Tokyo) 1976; 80: 87581. Kanematsu T, Takenaka K, Matsumata T, Furuta T, Sugimachi K, Inokuchi K. Limited hepatic resection effective for selected cirrhotic patients with primary liver cancer. Ann Surg 1984; 199 (1): 51-6. Kato A, Bamba H, Shanohara M, Yamauchi A, Ota S, Kawamoto C, Yoshida Y. Relationship between expression of cyclin D1 and impaired liver regeneration observed in fibrotic or cirrhotic rats. J Gastroenterol Hepatol 2005; 20: 1198-205. Krahenbuhl S, Reichen J. Adaptation of mitochondrial metabolism in liver cirrhosis. Different strategies to maintain a vital function. Scand J Gastroenterol Suppl 1992; 193: 90-6. Krahenbuhl S, Stucki J, Reichen J. Mitochondrial function in carbon tetrachlorideinduced cirrhosis in the rat. Qualitative and quantitative defects. Biochem Pharmacol 1989; 38: 1583-8. Kubbutat MH, Jones SN, Vousden KH. Regulation of p53 stability by Mdm2. Nature 1997; 387: 299-303. Kubin T, Gohda E, Yamamoto I. Expression of the protooncogene mdm2 markedly increases in response to carbon tetrachloride but not after partial hepatectomy in contrast to p53. Biochem Mol Biol Int 1995; 36:393-400. Kurokawa T, Nonami T, Sugiyama S, Ozawa T, Takagi H. Effects of long-acting superoxide dismutase on liver metabolism and major hepatic resection in rats with cirrhosis. Eur Surg Res 1991; 23: 65-72. LaBaer J, Garrett MD, Stevenson LF, Slingerland JM, Sandhu C, Chou HS, Fattaey A, Harlow E. New functional activities for the p21 family of CDK inhibitors. Genes Dev 1997; 11: 847-62. Lash LH, Putt DA, Matherly LH. Protection of NRK-52E cells, a rat renal proximal tubular cell line, from chemical-induced apoptosis by overexpression of a mitochondrial glutathione transporter. J Pharmacol Exp Ther 2002; 303: 476-86. 141 Lee F, Li Y, Zhu H, Yang S, Lin H, Trush M, Diehl A. Tumor necrosis factor increase mitochondrial oxidant production and induces expression of uncoupling protein-2 in the regenerating mice liver. Hepatology 1999; 29: 677-87. Lindroos PM, Zarnegar R, Michalopoulos GK. Hepatocyte growth factor (hepatopoietin A) rapidly increases in plasma before DNA synthesis and liver regeneration stimulated by partial hepatectomy and carbon tetrachloride administration. Hepatology 1991; 13: 74350. Li X, Benjamin IS, Alexander B. Reproducible production of thioacetamide-induced macronodular cirrhosis in the rat with no mortality. J Hepatol. 2002; 36: 488-93. Low TY, Leow CK, Salto-Tellez M, Chung MC. A proteomic analysis of thioacetamideinduced hepatotoxicity and cirrhosis in rat livers. Proteomics 2004; 4: 3960-74. Loyer P, Glaise D, Cariou S, Baffet G, Meijer L, Guguen-Guillouzo C. Expression and activation of cdks (1 and 2) and cyclins in the cell cycle progression during liver regeneration. J Biol Chem. 1994; 269: 2491-500. Lu XP, Koch KS, Lew DJ, Dulic V, Pines J, Reed SI, Hunter T, Leffert HL. Induction of cyclin mRNA and cyclin-associated histone H1 kinase during liver regeneration. J Biol Chem 1992; 267: 2841-4. Mann D, Lam W, Hjelm M, So N, Yeung D, Metreweli C, Lau W. Human liver regeneration: Hepatic energy economy is less efficient when the organ is diseased. Hepatol 2001; 34: 557-65. Martensson J, Lai JC, Meister A. High-affinity transport of glutathione is part of a multicomponent system essential for mitochondrial function. Proc Natl Acad Sci U S A 1990; 87: 7185-9. Masson S, Scotte M, Francois A, Coeffier M, Provot F, Hiron M, Teniere P, Fallu J, Salier JP, Daveau M. Changes in growth factor and cytokine mRNA levels after hepatectomy in rat with CCl4-induced cirrhosis. Am J Physiol 1999; 277: G838-46. Martin-Castellanos C, Moreno S. Recent advances on cyclins, CDKs and CDK inhibitors. Trends Cell Biol 1997; 7: 95-8. Massague J. Transforming growth factor-alpha. A model for membrane-anchored growth factors. J Biol Chem 1990; 265: 21393-6. Matsunami H, Kawasaki S, Ishizone S, Hashikura Y, Ikegami T, Makuuchi M, Kawarasaki H, Iwanaka T, Nose A, Takemura M. Serial changes of h-HGF and IL-6 in living-related donor liver transplantation with special reference to their relationship to intraoperative portal blood flow. Transplant Proc 1992; 24: 1971-2. 142 Meister A. Glutathione, ascorbate, and cellular protection. Cancer Res 1994; 54: 196975s. Meredith MJ, Reed DJ. Status of the mitochondrial pool of glutathione in the isolated hepatocyte. J Biol Chem 1982; 257: 3747-53. Messina J, Lawrence D. Cell cycle progression of glutathioine-depleted human peripheral blood mononuclear cells is inhibited at S phase. J Immunol 1989; 143: 1974-81. Metcalfe AM, Dixon RM, Radda GK. Wild-type but not mutant p53 activates the hepatocyte growth factor/scatter factor promoter. Nucleic Acids Res 1997; 25: 983-6. Michalopoulos GK and DeFrances MC. Liver regeneration. Science 1997; 276: 60-5. Milani S, Herbst H, Schuppan D, Grappone C, Pellegrini G, Pinzani M, Casini A, Calabro A, Ciancio G, Stefanini F, et al. Differential expression of matrixmetalloproteinase-1 and -2 genes in normal and fibrotic human liver. Am J Pathol 1994; 144: 528-37. Mitchell JB, Russo A. The role of glutathione in radiation and drug induced cytotoxicity. Br J Cancer Suppl 1987; 8: 96-104. Mitsui S, Ohuchi A, Adachi-Yamada T, Hotta M, Tsuboi R, Ogawa H. Cyclin-dependent kinase inhibitors, p21(waf1/cip1) and p27(kip1), are expressed site- and hair cycledependently in rat hair follicles. J Dermatol Sci 2001; 25: 164-9. Milani S, Herbst H, Schuppan D, Stein H, Surrenti C. Transforming growth factors beta and beta are differentially expressed in fibrotic liver disease. Am J Pathol 1991; 139: 1221-9. Miyamoto Y, Koh Y, Park Y, Fujiwara N, Sakiyama H, Misonou Y, Ookawara T, Suzuki K, Honke K, Taniguchi N. Oxidative stress caused by inactivation of glutathione peroxidase and adaptive responses. Biol Chem 2003; 384: 567-74. Moller B, Dargel R. Structural and functional impairment of mitochondria from rat livers chronically injured by thioacetamide. Acta Pharmacol Toxicol (Copenh). 1984; 55 (2): 126-32. Morgan DO. Cyclin dependent kinase: engines, clocks, and microprocessors. Annu Rev Cell Dev Biol 1997; 13: 261-91. Morimoto T, Isselhard W. Absence of mitochondrial enhancement in the remnant liver after partial hepatectomy in cirrhotic rats. Res Exp Med (Berl) 1992; 192: 89-97. 143 Muller D, Sommer M, Kretzschmar M, Zimmermann T, Buko V, Lukivskaya O, Dargel R. Lipid peroxidation in thioacetamide-induced macronodular rat liver cirrhosis. Arch Toxicol 1991; 65: 199-203. Murakami M, Hibi M, Nakagawa N, Nakagawa T, Yasukawa K, Yamanishi K, Taga T, Kishimoto T. IL-6-induced homodimerization of gp130 and associated activation of a tyrosine kinase. Science 1993; 260: 1808-10. Murray AW. Recycling the cell cycle: cyclins revisited. Cell 2004; 116: 221-34. Nakamura T, Nishizawa T, Hagiya M, Seki T, Shimonishi M, Sugimura A, Tashiro K, Shimizu S. Molecular cloning and expression of human hepatocyte growth factor. Nature 1989; 342: 440-3. Nakamura T, Nawa K, Ichihara A, Kaise N, Nishino T. Purification and subunit structure of hepatocyte growth factor from rat platelets. FEBS Lett 1987; 224: 311-6. Nakamura T, Nawa K, Ichihara A. Partial purification and characterization of hepatocyte growth factor from serum of hepatectomized rats. Biochem Biophys Res Commun 1984; 122: 1450-9. Nakanishi M, Adami GR, Robetorye RS, Noda A, Venable SF, Dimitrov D, PereiraSmith OM, Smith JR. Exit from G0 and entry into the cell cycle of cells expressing p21Sdi1 antisense RNA. Proc Natl Acad Sci U S A 1995; 92: 4352-6. Napoli J, Prentice D, Niinami C, Bishop GA, Desmond P, McCaughan GW. Sequential increases in the intrahepatic expression of epidermal growth factor, basic fibroblast growth factor, and transforming growth factor beta in a bile duct ligated rat model of cirrhosis. Hepatology 1997; 26: 624-33. Nozu F, Takeyama N, Tanaka T. Changes of hepatic fatty acid metabolism produced by chronic thioacetamide administration in rats. Hepatology 1992; 15: 1099-1106. Ohkawa H, Ohishi N, Yagi K. Assays for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351-8. Ohtsubo M, Theodoras AM, Schumacher J, Roberts JM, Pagano M. Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol 1995; 15: 261224. Ozdogan M, Ersoy E, Dundar K, Albayrak L, Devay S. Beneficial effect of hyperbaric oxygenation on liver regeneration in cirrhosis. J Surg Res 2005; 129: 260-4. Paradis V, Mathurin P, Kollinger M, Imbert-Bismut F, Charlotte F, Piton A, Opolon P, Holstege A, Poynard T, Bedossa P. In situ detection of lipid peroxidation in chronic hepatitis C: correlation with pathological features. J Clin Pathol 1997; 50 (5): 401-6. 144 Parkin DM, Bray FI, Devesa SS. Cancer burden in the year 2000. The global picture. Eur J Cancer 2001; 37 Suppl 8: S4-66. Penney C, Bolger G. A simple microassay for inorganic phosphate, II. Anal Biochem 1978; 89: 297-303. Pines J. Cyclins and cyclin-dependent kinases: a biochemical view. Biochem J 1995; 308: 697-711. Poot M, Teubert H, Rabinovitch P, Kavanagh T. De novo synthesis of glutathione is required for both entry into and progression through the cell cycle. J Cell Physiol 1995; 163: 555-60. Proctor E, Chatamra K. High yield micronodular cirrhosis in the rat. Gastroenterology 1982; 83: 1183-90. Pujol MJ, Jaime M, Serratosa J, Jaumot M, Agell N, Bachs O. Differential association of p21Cip1 and p27Kip1 with cyclin E-CDK2 during rat liver regeneration. J Hepatol 2000; 33: 266-74. Racker E. Glutathione reductase from bakers’ yeast and beef liver. J Biol Chem 1955; 217: 855-65. Rai RM, Yang SQ, McClain C, Karp CL, Klein AS, Diehl AM. Kupffer cell depletion by gadolinium chloride enhances liver regeneration after partial hepatectomy in rats. Am J Physiol 1996; 270: G909-18. Rana B, Xie Y, Mischoulon D, Bucher NL, Farmer SR. The DNA binding activity of C/EBP transcription factor is regulated in the G1 phase of the hepatocyte cell cycle. J Biol Chem 1995; 270: 18123-32. Rastogi VK, Girvin ME. Structural changes linked to proton translocation by subunit c of the ATP synthase. Nature 1999; 402: 263-8. Rininger JA, Goldsworthy TL, Babish JG. Time course comparison of cell-cycle protein expression following partial hepatectomy and WY14,643-induced hepatic cell proliferation in F344 rats. Carcinogenesis 1997; 18: 935-41. Russell WE, Carver RS. The EGF/TGFalpha family of growth factors and their receptors. In: Strain AJ, Diehl AM, editors. Liver Growth and Repair. London: Chapman & Hall; 1998. p. 185-218. Russell WE, Kaufman WK, Sitaric S, Luetteke NC, Lee DC. Liver regeneration and hepatocarcinogenesis in transforming growth factor-alpha-targeted mice. Mol Carcinog 1996; 15: 183-9. 145 Russo AA, Jeffrey PD, Patten AK, Massague J, Pavletich NP. Crystal structure of the p27Kip1 cyclin-dependent-kinase inhibitor bound to the cyclin A-Cdk2 complex. Nature 1996; 382: 325-31. Ruwart MJ, Wilkinson KF, Rush BD, Vidmar TJ, Peters KM, Henley KS, Appelman HD, Kim KY, Schuppan D, Hahn EG. The integrated value of serum procollagen III peptide over time predicts hepatic hydroxyproline content and stainable collagen in a model of dietary cirrhosis in the rat. Hepatology 1989; 10: 801-6. Sanz N, Diez-Fernandez C, Fernandez-Simon L, Alvarez A, Cascales M. Relationship between antioxidant systems, intracellular thiols and DNA ploidy in liver of rats during experimental cirrhogenesis. Carcinogenesis 1995; 16: 1585-93. Saraste M. Oxidative phosphorylation at the fin de siecle. Science 1999; 283 (5407): 1488-93. Satoh S, Tanaka A, Hatano E, Inomoto T, Iwata S, Kitai T, Shinohara H, Tsunekawa S, Chance B, Yamaoka Y. Energy metabolism and regeneration in transgenic mouse liver expressing creatine kinase after major hepatectomy. Gastroenterology 1996; 110: 116674. Schmidt C, Bladt F, Goedecke S, Brinkmann V, Zschiesche W, Sharpe M, Gherardi E, Birchmeier C. Scatter factor/hepatocyte growth factor is essential for liver development. Nature 1995; 373: 699-702. Sherr CJ. Cancer cell cycles. Science 1996; 274: 1672-7. Sherr CJ, Roberts JM. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev 1995; 9:1149-63. Sherr CJ. G1 phase progression: cycling on cue. Cell 1994; 79: 551-5. Shigenaga MK, Hagen TM, Ames BN. Oxidative damage and mitochondrial decay in aging. Proc Natl Acad Sci U S A 1994; 91: 10771-8. Shin TH, Paterson AJ, Kudlow JE. p53 stimulates transcription from the human transforming growth factor alpha promoter: a potential growth-stimulatory role for p53. Mol Cell Biol 1995; 15: 4694-701. Shurety W, Merino-Trigo A, Brown D, Hume DA, Stow JL. Localization and post-Golgi trafficking of tumor necrosis factor-alpha in macrophages. J Interferon Cytokine Res 2000; 20: 427-38. 146 Skehel JM, Fearnley IM, Walker JE. NADH:ubiquinone oxidoreductase from bovine heart mitochondria: sequence of a novel 17.2-kDa subunit. FEBS Lett 1998; 438 (3): 301-5. Soboll S, Stucki J. Regulation of the degree of coupling of oxidative phosphorylation in intact rat liver. Biochim Biophys Acta 1985; 807: 245-54. Stadtman ER. Protein oxidation and aging. Science 1992; 257: 1220-4. Streetz KL, Luedde T, Manns MP, Trautwein C. Interleukin and liver regeneration. Gut. 2000; 47: 309-12. Tashiro K, Hagiya M, Nishizawa T, Seki T, Shimonishi M, Shimizu S, Nakamura T. Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues. Proc Natl Acad Sci U S A 1990; 87: 3200-4. Taub R. Liver regeneration: from myth to mechanism. Nat Rev Mol Cell Biol. 2004; 5: 836-47. Tewari M, Dobrzanski P, Mohn KL, Cressman DE, Hsu JC, Bravo R, Taub R. Rapid induction in regenerating liver of RL/IF-1 (an I kappa B that inhibits NF-kappa B, RelBp50, and c-Rel-p50) and PHF, a novel kappa B site-binding complex. Mol Cell Biol 1992; 12: 2898-908. Tomiya T, Ogata I, Yamaoka M, Yanase M, Inoue Y, Fujiwara K. The mitogenic activity of hepatocyte growth factor on rat hepatocytes is dependent upon endogenous transforming growth factor-alpha. Am J Pathol 2000; 157: 1693-701. Tovey MG, Gugenheim J, Guymarho J, Blanchard B, Vanden Broecke C, Gresser I, Bismuth H, Reynes M. Genes for interleukin-1, interleukin-6, and tumor necrosis factor are expressed at markedly reduced levels in the livers of patients with severe liver disease. Autoimmunity 1991; 10: 297-310. Trembley JH, Kren BT, Steer CJ. Posttranscriptional regulation of cyclin B messenger RNA expression in the regenerating rat liver. Cell Growth Differ 1994; 5: 99-108. Trumpower BL. The protonmotive Q cycle. Energy transduction by coupling of proton translocation to electron transfer by the cytochrome bc1 complex. J Biol Chem 1990; 265 (20): 11409-12. Tsai J, King K, Chang C, Wei Y. Changes of mitochondrial respiratory functions and superoxide dismutase activity during liver regeneration. Biochem Internat 1992; 28: 205217. Tsukamoto H, Matsuoka M, French SW. Experimental models of hepatic fibrosis: a review. Semin Liver Dis 1990; 10: 56-65. 147 Tsukihara T, Aoyama H, Yamashita E, Tomizaki T, Yamaguchi H, Shinzawa-Itoh K, Nakashima R, Yaono R, Yoshikawa S. The whole structure of the 13-subunit oxidized cytochrome c oxidase at 2.8 A. Science 1996; 272 (5265): 1136-44. Turrens JF, Alexandre A, Lehninger AL. Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria. Arch Biochem Biophys 1985; 237 (2): 408-14. Turrens JF, Boveris A. Generation of superoxide anion by the NADH dehydrogenase of bovine heart mitochondria. Biochem J 1980; 191 (2): 421-7. Uehyara Y, Minowa O, Mori C, Shiota K, Kuno J, Noda T, Kitamura N. Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor. Nature 1995; 373: 702-5. Vendemiale G, Guerrieri F, Grattagliano I, Didonna D, Muolo L, Altomare E. Mitochondrial oxidative phosphorylation and intracellular glutathione compartmentation during rat liver regeneration. Hepatology 1995; 21 (5): 1450-4. Vermeulen K, Van Bockstaele DR, Berneman ZN. The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif 2003; 36: 131-49. Walker DH, Maller JL. Role for cyclin A in the dependence of mitosis on completion of DNA replication. Nature 1991; 354: 314-7. Walker JE. The NADH:ubiquinone oxidoreductase (complex I) of respiratory chains. Q Rev Biophys 1992; 25 (3): 253-324. Wasser S, Tan CE. Experimental models of hepatic fibrosis in the rat. Ann Acad Med Singapore 1999; 28: 109-11. Webber EM, FitsGerald MJ, Brown PI, Bartlett MH, Fausto N. Transforming growth factor-alpha expression during liver regeneration after partial hepatectomy and toxic injury, and potential interactions between transforming growth factor-alpha and hepatocyte growth factor. Hepatology 1993; 18: 1422-31. Weiner FR, Giambrone MA, Czaja MJ, Shah A, Annoni G, Takahashi S, Eghbali M, Zern MA. Ito-cell gene expression and collagen regulation. Hepatology 1990; 11: 111-7. Weber J, Senior AE. Catalytic mechanism of F1-ATPase. Biochim Biophys Acta 1997; 1319: 19-58. Wikstrom MK. Proton pump coupled to cytochrome c oxidase in mitochondria. Nature 1977; 266 (5599): 271-3. 148 Woo RA, Poon RY. Cyclin-dependent kinases and S phase control in mammalian cells. Cell Cycle 2003; 2: 316-24. Wu J, Zern MA. Hepatic stellate cells: a target for the treatment of liver fibrosis. J Gastroenterol 2000; 35: 665-72. Yamada Y, Webber EM, Kirillova I, Peschon JJ, Fausto N. Analysis of liver regeneration in mice lacking type or type tumor necrosis factor receptor: requirement for type but not type receptor. Hepatology 1998; 28: 959-70. Yamada Y, Kirillova I, Peschon JJ, Fausto N. Initiation of liver growth by tumor necrosis factor: deficient liver regeneration in mice lacking type I tumor necrosis factor receptor. Proc Natl Acad Sci USA 1997; 94: 1441-6. Yanagida H, Kaibori M, Hijikawa T, Kwon AH, Kamiyama Y, Okumura T. Administration of rhHGF-activator via portal vein stimulates the regeneration of cirrhotic liver after partial hepatectomy in rats. J Surg Res 2006; 130: 38-44. Yang SQ, Lin HZ, Yin M, Albrecht JH, Diehl AM. Effects of chronic ethanol consumption on cytokine regulation of liver regeneration. Am J Physiol 1998; 275: G696-704. Zhao G, Nakano K, Chijiiwa K, Ueda J, Tanaka M. Inhibited activities in CCAAT/enhancer-binding protein, activating protein-1 and cyclins after hepatectomy in rats with thioacetamide-induced liver cirrhosis. Biochem Biophys Res Commun 2002; 292: 474-81. Zimmermann T, Muller A, Machnik G, Franke H, Schubert H, Dargel R. Biochemical and morphological studies on production and regression of experimental liver cirrhosis induced by thioacetamide in Uje: WIST rats. Z Versuchstierkd 1987; 30: 165-80. 149 [...]... Changes in liver mitochondrial state 3 respiration of control (solid line) and cirrhotic rats (broken lines) following partial hepatectomy 74 Fig 4.2B Changes in liver mitochondrial state 4 respiration of control (solid line) and cirrhotic rats (broken lines) following partial hepatectomy 75 Fig 4.2C Changes in liver mitochondrial respiratory control ratio (RCR) of control (solid line) and cirrhotic rats. .. (broken lines) following partial hepatectomy 76 Fig 4.2D Changes in liver mitochondrial ADP/O ratio of control (solid line) and cirrhotic rats (broken lines) following partial hepatectomy 77 Fig 4.3 Mitochondrial ATPase activities from control (solid line) and cirrhotic 11 rats (broken lines) after partial hepatectomy 78 Fig 5.1A Mitochondrial glutathione (GSH) content in control (solid line) and cirrhotic. .. PH on cyclin D3 expression in healthy rats (white bars) and cirrhotic rats (black bars) 109 Fig 6.8 Effect of PH on cyclin E expression in healthy rats (white bars) and cirrhotic rats (black bars) 111 Fig 6.9 Effect of PH on cyclin A expression in healthy rats (white bars) and cirrhotic rats (black bars) 112 Fig 6.10 Effect of PH on cdk4 expression in healthy rats (white bars) and cirrhotic rats (black... cirrhotic rats (broken lines) after partial hepatectomy 70 Fig 4.1B Time-dependent changes of NADH-cytochrome c reductase (NCCR) activities from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 71 Fig 4.1C Time-dependent changes of cytochrome c oxidase (CCO) activities from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 73 Fig 4.2A Changes. .. expression in normal and cirrhotic rats after partial hepatectomy 95 Fig 5.6 Mitochondrial TBARS level from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 97 Fig 6.1 Hepatic TNF-α concentration after PH on healthy rats (white bars) and cirrhotic rats (black bars) 103 Fig 6.2 Hepatic IL-6 concentration after PH on healthy rats (white bars) and cirrhotic rats (black... capacity was also observed in cirrhotic rats with lower proliferating cell nuclear antigen labeling index and expression of G1 regulatory cell cycle-related proteins (Yanagida H et al, 2005; Ozdogan M et al, 2005; Kato A et al, 2005) In this study, we aim to examine the differences in the responses of normal and cirrhotic livers to partial hepatectomy in relation to the factors influencing liver regeneration... reductase (GRd) activity from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 92 Fig 5.4B Cytosolic glutathione reductase (GRd) activity from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 93 Fig 5.5A Western blot analysis of mitochondrial catalase expression in normal and cirrhotic rats after partial hepatectomy 94 Fig 5.5B Western blot analysis... activity from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 88 Fig 5.3A Mitochondrial glutathione peroxidase (GPx) activity from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 90 Fig 5.3B Cytosolic glutathione peroxidase (GPx) activity from control (solid line) and cirrhotic rats (broken lines) after partial hepatectomy 91 Fig 5.4A Mitochondrial... Amino acids Lipids Fatty acids Cholesterol Lipidproteins Fat-soluble vitamins Water-soluble vitamins Inactivation of various substances Toxins Steroids Other hormones Synthesis of plasma proteins Acute-phase proteins Albumin Clotting factors Steroid-binding and other hormone-binding proteins Immunity Kupffer cells 18 Growth Factors (HGF, TGFα) S Cytokines TNF IL-6 G0 G1 G2 M Fig 1.1 Multistep model of... (M) phase During interphase, the cell synthesizes RNA, produces proteins and grows in size Interphase can be divided into Gap 0 (G0), Gap 1 (G1), synthesis (S) phase and Gap 2 (G2) In G0, the cell leaves the cycle and quits dividing In G1, the cell increases in size, produces RNA and synthesizes proteins There is a checkpoint in G1 to ensure that the cell is ready for DNA synthesis During S phase, . their understanding and support throughout my study. 1 CELLULAR CHANGES POST HEPATECTOMY IN CIRRHOTIC VS NORMAL RATS TABLE OF CONTENTS CHAPTER 1 INTRODUCTION . capacity following partial hepatectomy 124 7.5 Expression of cyclins, cdks and cdk inhibitors in normal and cirrhotic livers 126 7.6 Changes in expression of cyclins, cdks and cdk inhibitors. (solid line) and cirrhotic rats (broken lines) following partial hepatectomy 75 Fig 4.2C Changes in liver mitochondrial respiratory control ratio (RCR) of control (solid line) and cirrhotic rats