Promotion of mitochondrial energy metabolism during hepatocyte apoptosis in a rat model of acute liver failure

Chen,Li‑Yan; Yang,Baoshan; Zhou,Li; Ren,Feng; Duan,Zhong‑Ping; Ma,Ying‑Ji

doi:10.3892/mmr.2015.4029

Promotion of mitochondrial energy metabolism during hepatocyte apoptosis in a rat model of acute liver failure

Authors:
- Li‑Yan Chen
- Baoshan Yang
- Li Zhou
- Feng Ren
- Zhong‑Ping Duan
- Ying‑Ji Ma
View Affiliations

Affiliations: The Second Department of Infectious Diseases, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China, Beijing Artificial Liver Treatment and Training Center, Beijing Youan Hospital, Capital Medical University, Beijing 100069, P.R. China, The Fourth Department of Infectious Diseases, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: July 2, 2015 https://doi.org/10.3892/mmr.2015.4029
Pages: 5035-5041
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hepatocyte apoptosis and energy metabolism in mitochondria have an important role in the mechanism of acute liver failure (ALF). However, data on the association between apoptosis and the energy metabolism of hepatocytes are lacking. The current study assessed the activity of several key enzymes in mitochondria during ALF, including citrate synthase (CS), carnitine palmitoyltransferase‑1 (CPT‑1) and cytochrome c oxidase (COX), which are involved in hepatocyte energy metabolism. A total of 40 male Sprague‑Dawley rats were divided into five groups and administered D‑galactosamine and lipopolysaccharide to induce ALF. Hepatic pathology and terminal deoxynucleotidyl transferase‑mediated dUTP nick end labeling examinations indicated that hepatocyte apoptosis was observed at 4 h and increased 8 h after ALF. Hepatocyte necrosis appeared at 12 h and was significantly higher at 24 h with inflammatory cell invasion. The results measured by electron microscopy indicated that ultrastructural changes in mitochondria began at 4 h and the mitochondrial outer membrane was completely disrupted at 24 h resulting in mitochondrial collapse. The expression of CS, CPT‑1 and COX was measured and analyzed using assay kits. The activity and protein expression of CS, CPT‑1 and COX began to increase at 4 h, reached a peak at 8 h and decreased at 12 h during ALF. The activities of CS, CPT‑1 and COX were enhanced during hepatocyte apoptosis suggesting that these enzymes are involved in the initiation and development of ALF. Therefore, these results demonstrated that energy metabolism is important in hepatocyte apoptosis during ALF and hepatocyte apoptosis is an active and energy‑consuming procedure. The current study on how hepatocyte energy metabolism affects the transmission of death signals may provide a basis for the early diagnosis and development of an improved therapeutic strategy for ALF.

View Figures

View References

1	Liver Failure and Artificial Liver Group; Chinese Society of Infectious Diseases; Chinese Medical Association; Severe Liver Diseases and Artificial Liver Group: Chinese Society of Hepatology, Chinese Medical Association: Diagnostic and treatment guidelines for liver failure. Zhonghua Gan Zang Bing Za Zhi. 14:643–646. 2006.In Chinese.
2	Orrenius S: Reactive oxygen species in mitochondria-mediated cell death. Drug Metab Rev. 39:443–455. 2007. View Article : Google Scholar : PubMed/NCBI
3	Kushnareva Y and Newmeyer DD: Bioenergetics and cell death. Ann NY Acad Sci. 1201:50–57. 2010. View Article : Google Scholar : PubMed/NCBI
4	Ryu SY, Peixoto PM, Teijido O, Dejean LM and Kinnally KW: Role of mitochondrial ion channels in cell death. Biofactors. 36:255–263. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ohta S: A multi-functional organelle mitochondrion is involved in cell death, proliferation and disease. Curr Med Chem. 10:2485–2494. 2003. View Article : Google Scholar : PubMed/NCBI
6	Zhao BC: Role of liver in lipid metabolism. Biochemistry (Chin). Zhou AR: 18. 5th edition. People's Medical Publishing House; Beijing: pp. 3652003
7	Zhao BC: Regulation of activities of enzymes. Biochemistry (Chin). Zhou AR: 4. 5th edition. People's Medical Publishing House; Beijing: pp. 63–64. 2003
8	Wiegand G, Remington S, Deisenhofer J and Huber R: Crystal structure analysis and molecular model of a complex of citrate synthase with oxaloacetate and S-acetonyl-coenzyme. A J Mol Biol. 174:205–219. 1984. View Article : Google Scholar
9	Cheng TL, Liao CC, Tsai WH, Lin CC, Yeh CW, Teng CF and Chang WT: Identification and characterization of the mitochondrial targeting sequence and mechanism in human citrate synthase. J Cell Biochem. 107:1002–1015. 2009. View Article : Google Scholar : PubMed/NCBI
10	Donald LJ and Duckworth HW: Molecular cloning of the structural gene for Acinetobacter citrate synthase. Biochem Biophys Res Commun. 141:797–803. 1986. View Article : Google Scholar : PubMed/NCBI
11	Bloxham DP, Parmelee DC, Kumar S, Wade RD, Ericsson LH, Neurath H, Walsh KA and Titani K: Primary structure of porcine heart citrate synthase. Proc Natl Acad Sci USA. 78:5381–5385. 1981. View Article : Google Scholar : PubMed/NCBI
12	Kerner J and Hoppel C: Fatty acid import into mitochondria. Biochim Biophys Acta. 1486:1–17. 2000. View Article : Google Scholar : PubMed/NCBI
13	Ostermeier C, Iwata S and Michel H: Cytochrome c oxidase. Curr Opin Struct Biol. 6:460–466. 1996. View Article : Google Scholar : PubMed/NCBI
14	Kadenbach B, Barth J, Akgün R, Freund R, Linder D and Possekel S: Regulation of mitochondrial energy generation in health and disease. Biochim Biophys Acta. 1271:103–109. 1995. View Article : Google Scholar : PubMed/NCBI
15	Kadenbach B, Hüttemann M, Arnold S, Lee I and Bender E: Mitochondrial energy metabolism is regulated via nuclear-coded subunits of cytochrome c oxidase. Free Radic Biol Med. 29:211–221. 2000. View Article : Google Scholar : PubMed/NCBI
16	Jezek P and Hlavatá L: Mitochondria in homeostasis of reactive oxygen species in cell, tissues and organism. Int J Biochem Cell Biol. 37:2478–2503. 2005. View Article : Google Scholar : PubMed/NCBI
17	Feldstein AE, Werneburg NW, Canbay A, Guicciardi ME, Bronk SF, Rydzewski R, Burgart LJ and Gores GJ: Free fatty acids promote hepatic lipotoxicity by stimulating TNF-alpha expression via a lysosomal pathway. Hepatology. 40:185–194. 2004. View Article : Google Scholar : PubMed/NCBI
18	Malhi H, Bronk SF, Werneburg NW and Gores GJ: Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis. J Biol Chem. 281:12093–12101. 2006. View Article : Google Scholar : PubMed/NCBI
19	Turrens JF: Mitochondrial formation of reactive oxygen species. J Physiol. 552:335–344. 2003. View Article : Google Scholar : PubMed/NCBI
20	Hsieh CL, Huang CN, Lin YC and Peng RY: Molecular action mechanism against apoptosis by aqueous extract from guava budding leaves elucidated with human umbilical vein endothelial cell (HUVEC) model. J Agric Food Chem. 55:8523–8533. 2007. View Article : Google Scholar : PubMed/NCBI
21	Rana SV: Metals and apoptosis: Recent developments. J Trace Elem Med Biol. 22:262–284. 2008. View Article : Google Scholar : PubMed/NCBI
22	Giordano A, Calvani M, Petillo O, Grippo P, Tuccillo F, Melone MA, Bonelli P, Calarco A and Peluso G: tBid induces alterations of mitochondrial fatty acid oxidation flux by malonyl-CoA-independent inhibition of carnitine palmitoyl-transferase-1. Cell Death Differ. 12:603–613. 2005. View Article : Google Scholar : PubMed/NCBI
23	Harada H and Grant S: Apoptosis regulators. Rev Clin Exp Hematol. 7:117–138. 2003.
24	Deutschman CS, Haber BA, Andrejko K, Cressman DE, Harrison R, Elenko E and Taub R: Increased expression of cytokine-induced neutrophil chemoattractant in septic rat liver. Am J Physiol. 271:R593–R600. 1996.PubMed/NCBI
25	Decker K and Keppler D: Galactosamine hepatitis: Key role of the nucleotide deficiency period in the pathogenesis of cell injury and cell death. Rev Physiol Biochem Pharmacol. 71:77–106. 1974. View Article : Google Scholar : PubMed/NCBI
26	Qiu Z, Kwon AH, Tsuji K, Kamiyama Y, Okumura T and Hirao Y: Fibronectin prevents D-galactosamine/lipopolysac-charide-induced lethal hepatic failure in mice. Shock. 25:80–87. 2006. View Article : Google Scholar
27	Canbay A, Friedman S and Gores GJ: Apoptosis: The nexus of liver injury and fibrosis. Hepatology. 39:273–278. 2004. View Article : Google Scholar : PubMed/NCBI
28	Malhi H, Gores GJ and Lemasters JJ: Apoptosis and necrosis in the liver: A tale of two deaths? Hepatology. 43(2 Suppl 1): pp. S31–S44. 2006, View Article : Google Scholar
29	Vaquero J and Blei AT: Etiology and management of fulminant hepatic failure. Curr Gastroenterol Rep. 5:39–47. 2003. View Article : Google Scholar : PubMed/NCBI
30	Van Herreweghe F, Festjens N, Declercq W and Vandenabeele P: Tumor necrosis factor-mediated cell death: To break or to burst, that's the question. Cell Mol Life Sci. 67:1567–1579. 2010. View Article : Google Scholar : PubMed/NCBI
31	Davis CW, Hawkins BJ, Ramasamy S, Irrinki KM, Cameron BA, Islam K, Daswani VP, Doonan PJ, Manevich Y and Madesh M: Nitration of the mitochondrial complex I subunit NDUFB8 elicits RIP1- and RIP3-mediated necrosis. Free Radic Biol Med. 48:306–317. 2010. View Article : Google Scholar
32	Zhang DW, Shao J, Lin J, Zhang N, Lu BJ, Lin SC, Dong MQ and Han J: RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science. 325:332–336. 2009. View Article : Google Scholar : PubMed/NCBI
33	Clemmesen O: Splanchnic circulation and metabolism in patients with acute liver failure. Dan Med Bull. 49:177–193. 2002.PubMed/NCBI
34	Saibara T, Maeda T, Onishi S and Yamamoto Y: Plasma exchange and the arterial blood ketone body ratio in patients with acute hepatic failure. J Hepatol. 20:617–622. 1994. View Article : Google Scholar : PubMed/NCBI
35	Lee SJ, Kwon CH and Kim YK: Alterations in membrane transport function and cell viability induced by ATP depletion in primary cultured rabbit renal proximal tubular cells. Korean J Physiol Pharmacol. 13:15–22. 2009. View Article : Google Scholar : PubMed/NCBI
36	Zischka H, Larochette N, Hoffmann F, Hamöller D, Jägemann N, Lichtmannegger J, Jennen L, Müller-Höcker J, Roggel F, Göttlicher M, et al: Electrophoretic analysis of the mitochondrial outer membrane rupture induced by permeability transition. Anal Chem. 80:5051–5058. 2008. View Article : Google Scholar : PubMed/NCBI
37	Marín-García J, Goldenthal MJ, Damle S, Pi Y and Moe GW: Regional distribution of mitochondrial dysfunction and apoptotic remodeling in pacing-induced heart failure. J Card Fail. 15:700–708. 2009. View Article : Google Scholar : PubMed/NCBI
38	Chinta SJ, Rane A, Yadava N, Andersen JK, Nicholls DG and Polster BM: Reactive oxygen species regulation by AIF- and complex I-depleted brain mitochondria. Free Radic Biol Med. 46:939–947. 2009. View Article : Google Scholar : PubMed/NCBI
39	Puig T, Relat J, Marrero PF, Haro D, Brunet J and Colomer R: Green tea catechin inhibits fatty acid synthase without stimulating carnitine palmitoyltransferase-1 or inducing weight loss in experimental animals. Anticancer Res. 28:3671–3676. 2008.
40	Mazzarelli P, Pucci S, Bonanno E, Sesti F, Calvani M and Spagnoli LG: Carnitine palmitoyltransferase I in human carcinomas: A novel role in histone deacetylation? Cancer Biol Ther. 6:1606–1613. 2007. View Article : Google Scholar