Mitochondrial and bioenergetic dysfunction in human hepatic cells infected with dengue 2 virus.

Dengue virus infection affects millions of people all over the world. Although the clinical manifestations of dengue virus-induced diseases are known, the physiopathological mechanisms involved in deteriorating cellular function are not yet understood. In this study we evaluated for the first time the associations between dengue virus-induced cell death and mitochondrial function in HepG2, a human hepatoma cell line. Dengue virus infection promoted changes in mitochondrial bioenergetics, such as an increase in cellular respiration and a decrease in DeltaPsim. These alterations culminated in a 20% decrease in ATP content and a 15% decrease in the energy charge of virus-infected cells. Additionally, virus-infected cells showed several ultrastructural alterations, including mitochondria swelling and other morphological changes typical of the apoptotic process. The alterations in mitochondrial physiology and energy homeostasis preceded cell death. These results indicate that HepG2 cells infected with dengue virus are under metabolic stress and that mitochondrial dysfunction and alterations in cellular ATP balance may be related to the pathogenesis of dengue virus infection.

[1]  M. Guzmán,et al.  Dengue and dengue hemorrhagic fever in the Americas: lessons and challenges. , 2003, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[2]  Dan Huang,et al.  Analysis of intracellular nucleoside triphosphate levels in normal and tumor cell lines by high-performance liquid chromatography. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[3]  G. Brown,et al.  Control of respiration and oxidative phosphorylation in isolated rat liver cells. , 1990, European journal of biochemistry.

[4]  P. Desprès,et al.  Dengue virus M protein contains a proapoptotic sequence referred to as ApoptoM. , 2003, The Journal of general virology.

[5]  Tatiana El-Bacha,et al.  Mayaro virus infection alters glucose metabolism in cultured cells through activation of the enzyme 6-phosphofructo 1-kinase , 2004, Molecular and Cellular Biochemistry.

[6]  G. M. Walton,et al.  Adenosine Triphosphate Conservation in Metabolic Regulation , 2001 .

[7]  P. Marianneau,et al.  Report of a fatal case of dengue infection with hepatitis: demonstration of dengue antigens in hepatocytes and liver apoptosis. , 1999, Human pathology.

[8]  P. Jansen-Dürr,et al.  Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. , 2004, The Biochemical journal.

[9]  C C Howe,et al.  Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. , 1980, Science.

[10]  D. Gubler.  The global emergence/resurgence of arboviral diseases as public health problems. , 2002, Archives of medical research.

[11]  Guido Kroemer,et al.  Mitochondrial control of cell death , 2000, Nature Medicine.

[12]  Juan-Pablo Hernández,et al.  Ultrastructural studies on dengue virus type 2 infection of cultured human monocytes , 2005, Virology Journal.

[13]  C. Thompson,et al.  Mitochondrial respiratory control is lost during growth factor deprivation , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Friedman.  Protein Synthesis Directed by an Arbovirus , 1968, Journal of virology.

[15]  Zong-yin Qiu,et al.  Effect of ecdysterone on glucose metabolism in vitro. , 2006, Life sciences.

[16]  Yi-Ling Lin,et al.  Flavivirus Activates Phosphatidylinositol 3-Kinase Signaling To Block Caspase-Dependent Apoptotic Cell Death at the Early Stage of Virus Infection , 2005, Journal of Virology.

[17]  E. C. Slater.  THE RESPIRATORY CHAIN AND OXIDATIVE PHOSPHORYLATION , 1972 .

[18]  P. Marianneau,et al.  Liver histopathology and biological correlates in five cases of fatal dengue fever in Vietnamese children , 2001, Virchows Archiv.

[19]  K. Raivio,et al.  Depletion of liver adenine nucleotides induced by D-fructose. Dose-dependence and specificity of the fructose effect. , 1969, Biochemical pharmacology.

[20]  G. Jan,et al.  Study of PTPC Composition during Apoptosis for Identification of Viral Protein Target , 2003, Annals of the New York Academy of Sciences.

[21]  D. Gubler,et al.  Dengue and Dengue Hemorrhagic Fever , 1998, Clinical Microbiology Reviews.

[22]  K. Wirtz,et al.  "Wages of fear": transient threefold decrease in intracellular ATP level imposes apoptosis. , 2004, Biochimica et biophysica acta.

[23]  M. Guzmán,et al.  Oxidative stress in adult dengue patients. , 2004, The American journal of tropical medicine and hygiene.

[24]  P. Marianneau,et al.  Differing infection patterns of dengue and yellow fever viruses in a human hepatoma cell line. , 1998, Journal of Infectious Diseases.

[25]  O. Barth.  Replication of dengue viruses in mosquito cell cultures--a model from ultrastructural observations. , 1992, Memórias do Instituto Oswaldo Cruz.

[26]  G. Ronquist,et al.  Coxsackie B3 myocarditis induces a decrease in energy charge and accumulation of hyaluronan in the mouse heart , 1993, European journal of clinical investigation.

[27]  G. M. Walton,et al.  Adenosine triphosphate conservation in metabolic regulation. Rat liver citrate cleavage enzyme. , 1967, The Journal of biological chemistry.

[28]  M. Ripoli,et al.  Mitochondrial dysfunction in hepatitis C virus infection. , 2006, Biochimica et biophysica acta.

[29]  Y. L. Lin,et al.  Involvement of oxidative stress, NF-IL-6, and RANTES expression in dengue-2-virus-infected human liver cells. , 2000, Virology.

[30]  C. Ho,et al.  Inactivation of coupled respiration of mitochondria by inorganic arsenate and partial restoration by ATP. , 1972, Biochemical and biophysical research communications.

[31]  P. Desprès,et al.  Expression of dengue ApoptoM sequence results in disruption of mitochondrial potential and caspase activation. , 2003, Biochimie.

[32]  A. Kosaki,et al.  Effect of dexamethasone on the alternative splicing of the insulin receptor mRNA and insulin action in HepG2 hepatoma cells. , 1993, The Journal of biological chemistry.

[33]  T. Smith,et al.  J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. , 1991, Biochemistry.

[34]  G. McFadden,et al.  Viruses and apoptosis: meddling with mitochondria. , 2001, Virology.

[35]  M Crompton,et al.  The mitochondrial permeability transition pore and its role in cell death. , 1999, The Biochemical journal.

[36]  Marco Novelli,et al.  Mitochondrial dysfunction in a long-term rodent model of sepsis and organ failure. , 2004, American journal of physiology. Regulatory, integrative and comparative physiology.

[37]  M. Gutman,et al.  Control of succinate dehydrogenase in mitochondria. , 1971, Biochemistry.

[38]  D. E. Atkinson.  Cellular Energy Metabolism and its Regulation , 1977 .

[39]  P. Desprès,et al.  Apoptotic cell death in response to dengue virus infection: the pathogenesis of dengue haemorrhagic fever revisited. , 1998, Clinical and diagnostic virology.

[40]  G. McFadden,et al.  Apoptosis: an innate immune response to virus infection. , 1999, Trends in microbiology.

[41]  P. Desprès,et al.  Mechanisms of dengue virus-induced cell death. , 2003, Advances in virus research.

[42]  A. Mustafa,et al.  Cytokine cascade in dengue hemorrhagic fever: implications for pathogenesis. , 2000, FEMS immunology and medical microbiology.

[43]  A. Cederbaum,et al.  Oxidative stress and cytotoxicity induced by ferric-nitrilotriacetate in HepG2 cells that express cytochrome P450 2E1. , 1998, Molecular pharmacology.

[44]  G. Áñez,et al.  Short report: increased level of serum nitric oxide in patients with dengue. , 2002, The American journal of tropical medicine and hygiene.

[45]  D. Gubler,et al.  Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses , 2004, Nature Medicine.

[46]  M. Singer,et al.  Mitochondrial dysfunction in patients with severe sepsis: an EPR interrogation of individual respiratory chain components. , 2006, Biochimica et biophysica acta.