Derangements of liver tissue bioenergetics in Concanavalin A-induced hepatitis

BMC Gastroenterology

Abdul-Kader Souid¹  Eric PK Mensah-Brown²  Allen Shahin³  Mariam Al-Shamsi³ 

[1] Department of Pediatrics, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, United Arab Emirates (UAE;Department of Anatomy, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, United Arab Emirates (UAE;Department of Immunology, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, United Arab Emirates (UAE

关键词: IFN-γ;    ALT;    AST;    Caspase;    Hepatitis;    Concanavalin A (ConA);   

Others  :  858242
DOI  :  10.1186/1471-230X-13-6

 received in 2012-07-02, accepted in 2012-12-13,  发布年份 2013

【 摘 要 】

Background

A novel in vitro system was employed to investigate liver tissue respiration (mitochondrial O₂ consumption) in mice treated with concanavalin A (Con A). This study aimed to investigate hepatocyte bioenergetics in this well-studied hepatitis model.

Methods

C57Bl/6 and C57Bl/6 IFN-γ^−/− mice were injected intravenously with 12 mg ConA/kg. Liver specimens were collected at various timepoints after injection and analyzed for cellular respiration and caspase activation. Serum was analyzed for interferon-gamma (IFN-γ) and aminotransferases. Fluorescence activated cell sorting analysis was used to determine the phenotype of infiltrating cells, and light and electron microscopy were used to monitor morphological changes. Phosphorescence analyzer that measured dissolved O₂ as function of time was used to evaluate respiration.

Results

In sealed vials, O₂ concentrations in solutions containing liver specimen and glucose declined linearly with time, confirming zero-order kinetics of hepatocyte respiration. O₂ consumption was inhibited by cyanide, confirming the oxidation occurred in the respiratory chain. Enhanced liver respiration (by ≈68%, p<0.02) was noted 3 hr after ConA treatment, and occurred in conjunction with limited cellular infiltrations around the blood vessels. Diminished respiration (by ≈30%, p=0.005) was noted 12 hr after ConA treatment, and occurred in conjunction with deranged mitochondria, areas of necrosis, and prominent infiltrations with immune cells, most significantly, CD3⁺NKT⁺ cells. Increases in intracellular caspase activity and serum IFN-γ and aminotransferase levels were noted 3 hr after ConA treatment and progressed with time. The above-noted changes were less pronounced in C57Bl/6 IFN-γ^−/− mice treated with ConA.

Conclusions

Based on these results, liver tissue bioenergetics is increased 3 hr after ConA exposure. This effect is driven by the pathogenesis of the disease, in which IFN-γ and other cytokines contribute to. Subsequent declines in liver bioenergetics appear to be a result of necrosis and active caspases targeting the mitochondria within hepatocytes.

【 授权许可】

   
2013 Al-Shamsi et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Tiegs G, Hentschel J, Wendel JA: T-Cell-dependent experimental liver injury in mice inducible by concanavalin A. J Clin Invest 1992, 90:196-203.
• [2]Tiegs G: Experimental hepatitis and the role of cytokines. Acta Gastroenterol Belg 1997, 60:176-179.
• [3]Takeda K, Hayakawa Y, Van Kaer L, Matsuda H, Yaqita H, Okumura K: Critical contribution of liver natural killer T-cells to a murine model of hepatitis. Proc Natl Acad Sci USA 2000, 97:5498-5503.
• [4]Leist M, Wendel A: A novel mechanism of murine hepatocyte death inducible by concanavalin A. Hepatology 1996, 25:948-959.
• [5]Margalit M, Abu Ghazala S, Alper R, Elinav E, Klein A, Doviner V, Sherman Y, Thalenfeld B, Engelhardt D, Rabbani E, Ilan Y: Glucocerbroside treatment ameliorates ConA hepatitis by inhibition of NK T lymphocytes. Am J Physiol 2005, 289:G917-G925.
• [6]Rahman TM, Hodgson HJ: Animal models of acute hepatic failure. Int J Exp Path 2000, 81:145-157.
• [7]Tao Z, Penefsky HS, Goodisman J, Souid AK: Caspase activation by cytotoxic drugs (the caspase storm). Mol Pharm 2007, 4:583-585.
• [8]Al-Shamsi M, Al-Samri M, Al-Salam S, Conca W, Benedict S, Sudhadevi M, Biradar A, Asefa T, Souid A: Biocompatibility study of mesoporous silicate particles with cellular bioenergetics in murine tissues. Chem Res Toxicol 2010, 11:1796-1805.
• [9]Al Samri MT, Al Shamsi M, Al-Salam S, Marzouqi F, Al Mansouri A, Al-Hammadi S, Balhaj G, Al Dawaar SK, Al Hanjeri RS, Benedict S, Sudhadevi M, Conca W, Penefsky HS, Souid AK: Measurement of oxygen consumption by murine tissues in vitro. J Pharmacol Toxicol Meth 2011, 63:196-204.
• [10]Shaban S, Marzouqi F, Al Mansouri A, Penefsky H, Souid AK: Oxygen measurements via phosphorescence. Computer Meth Programs Biomed 2010, 100:265-268.
• [11]Lo LW, Koch CJ, Wilson DF: Calibration of oxygen-dependent quenching of the phosphorescence of Pd-meso-tetra (4-carboxyphenyl) porphine: A phosphor with general application for measuring oxygen concentration in biological systems. Anal Biochem 1996, 236:153-160.
• [12]Knolle PA, Gerke G, Löser E, Dienes HP, Gantner F, Tiegs G, Meyer zum Büschenfelde K, Lohse AW: Role of sinusoidal endothelial cells of the liver in Concanavalin A-induced hepatic injury in mice. Hepatology 1996, 24:824-829.
• [13]Sass G, Heinlein S, Agli A, Bang R, Schumann J, Tiegs G: Cytokine expression in three mouse models of experimental hepatitis. Cytokine 2002, 19:115-120.
• [14]Gottlier E, Van der Heiden MG, Thompson CB: Bcl-xL prevents the initial decrease in mitochondrial membrane potential and subsequent reactive oxygen species production during TNF α-induced apoptosis. Mol Cell Biol 2000, 20:5680-5689.
• [15]Tagawa YI, Sekikawa K, Iwakura Y: Suppression of ConA-induced hepatitis in IFN-γ−/− mice, but not in TNFα−/− mice. J Immunol 1997, 159:1418-1428.
• [16]Siebler J, Wirtz S, Klein S, Protschka M, Blessing M, Galle PR, Neurath MF: A key pathogenic role for the STAT1/T-bet signaling pathway in T-cell-mediated liver inflammation. Hepatology 2003, 38:1573-1580.
• [17]Jaruga B, Hong F, Kim WH, Gao B: IFN-γ/STAT1 acts as a pro-inflammatory signal in T-cell-mediated hepatitis via induction of multiple chemokines and adhesion molecules: A critical role of IRF-1. Am J Physiol 2004, 287:G1044-G1052.