Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

ACC 2024 Congress

Catch up on all the top stories and latest evidence in cardiology research with our ACC 2024 Congress hub page.
ADVANCED SEARCH
Log in
Top
BMC Gastroenterology
Published in: BMC Gastroenterology 1/2014

Open Access 01-12-2014 | Research article

Hyperammonia induces specific liver injury through an intrinsic Ca2+-independent apoptosis pathway

Authors: Jingjing Li, Zujiang Yu, Qiongye Wang, Duolu Li, Bin Jia, Yubing Zhou, Yanwei Ye, Shen Shen, Yanfang Wang, Shasha Li, Lu Bai, Quancheng Kan

Published in: BMC Gastroenterology | Issue 1/2014

Login to get access

Abstract

Background

Numerous pathological processes that affect liver function in patients with liver failure have been identified. Among them, hyperammonia is one of the most common phenomena.The purpose of this study was to determine whether hyperammonia could induced specific liver injury.

Methods

Hyperammonemic cells were established using NH4Cl. The cells were assessed by MTT, ELISA, and flow cytometric analyses. The expression levels of selected genes and proteins were confirmed by quantitative RT-PCR and western blot analyses.

Results

The effects of 20 mM NH4Cl pretreatment on the cell proliferation and apoptosis of primary hepatocytes and other cells were performed by MTT assays and flow cytometric analyses. Significant increasing in cytotoxicity and apoptosis were only observed in hepatocytes. The cell damage was reduced after adding BAPTA-AM but unchanged after adding EGTA. The expression levels of caspase-3, cytochrome C, calmodulin, and inducible nitric oxide synthase were increased and that of bcl-2 was reduced. The Na+-K+-ATPase activities in hyperammonia liver cells was no signiaficant difference compaired with the control group, but was decreased in astrocytes. NH4Cl pretreatment of primary hepatocytes promoted the activation of mitochondrial permeability transition pores and the mitochondria swelled irregularly.

Conclusions

Hyperammonia induces specific liver injury through an intrinsic Ca2+-independent apoptosis pathway.
Appendix
Available only for authorised users
Literature
1.
Ellis A, Wendon J: Circulatory, respiratory, cerebral, and renal derangements in acute liver failure: pathophysiology and management. Semin Liver Dis. 1996, 16: 379-388. 10.1055/s-2007-1007251.CrossRefPubMed
2.
Sandgren EP, Palmiter RD, Heckel JL, Sandgren EP, Palmiter RD, Heckel JL, Daugherty CC, Brinster RL, Degen JL: Complete hepatic regeneration after somatic deletion of an albumin-plasminogen activator transgene. Cell. 1991, 66: 245-256. 10.1016/0092-8674(91)90615-6.CrossRefPubMed
3.
Overturf K, Al-Dhalimy M, Finegold M, Grompe M: The repopulation potential of hepatocyte populations differing in size and prior mitotic expansion. Am J Pathol. 1999, 155: 2135-2143. 10.1016/S0002-9440(10)65531-9.CrossRefPubMedPubMedCentral
4.
Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Murase N, Boggs SS, Greenberger JS, Goff JP: Bone marrow as a potential source of hepatic oval cells. Science. 1999, 284: 1168-1170. 10.1126/science.284.5417.1168.CrossRefPubMed
5.
Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, Osborne L, Wang X, Finegold M, Weissman IL, Grompe M: Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med. 2000, 6: 1229-1234. 10.1038/81326.CrossRefPubMed
6.
7.
Taub R: Liver regeneration: from myth to mechanism. Nat Rev Mol Cell Biol. 2004, 5: 836-847. 10.1038/nrm1489.CrossRefPubMed
8.
Lange CM, Bojunga J, Hofmann WP, Wunder K, Mihm U, Zeuzem S, Sarrazin C: Severe lactic acidosis during treatment of chronic hepatitis B with entecavir in patients with impaired liver function. Hepatology. 2009, 50: 2001-2006. 10.1002/hep.23346.CrossRefPubMed
9.
Widmer R, Kaiser B, Engels M, Wunder K, Mihm U, Zeuzem S, Sarrazin C: Hyperammonemia causes protein oxidation and enhanced proteasomal activity in response to mitochondria-mediated oxidative stress in rat primary astrocytes. Arch Biochem Biophys. 2007, 464: 1-11. 10.1016/j.abb.2007.03.027.CrossRefPubMed
10.
Felipo V, Butterworth RF: Neurobiology of ammonia. Prog Neurobiol. 2002, 67: 259-279. 10.1016/S0301-0082(02)00019-9.CrossRefPubMed
11.
Drolz A, Jäger B, Wewalka M, Saxa R, Horvatits T, Roedl K, Perkmann T, Zauner C, Kramer L, Ferenci P, Fuhrmann V: Clinical impact of arterial ammonia levels in ICU patients with different liver diseases. Intensive Care Med. 2013, 39: 1227-1237. 10.1007/s00134-013-2926-8.CrossRefPubMed
12.
Skowronska M, Albrecht J: Alterations of blood brain barrier function in hyperammonemia: an overview. Neurotox Res. 2012, 21: 236-244. 10.1007/s12640-011-9269-4.CrossRefPubMed
13.
Rao KV, Norenberg MD: Cerebral energy metabolism in hepatic encephalopathy and hyperammonemia. Metab Brain Dis. 2001, 16: 67-78. 10.1023/A:1011666612822.CrossRefPubMed
14.
Rao VL, Murthy CR, Butterworth RF: Glutamatergic synaptic dysfunction in hyperammonemic syndromes. Metab Brain Dis. 1992, 7: 1-20. 10.1007/BF01000437.CrossRefPubMed
15.
Hazell AS, Butterworth RF: Hepatic encephalopathy: An update of pathophysiologic mechanisms. Proc Soc Exp Biol Med. 1999, 222: 99-112. 10.1046/j.1525-1373.1999.d01-120.x.CrossRefPubMed
16.
Kim IK, Niemi AK, Krueger C, Bonham CA, Concepcion W, Cowan TM, Enns GM, Esquivel CO: Liver transplantation for urea cycle disorders in pediatric patients: a single-center experience. Pediatr Transplant. 2013, 17: 158-167. 10.1111/petr.12041.CrossRefPubMed
17.
Adeva MM: Ammonium metabolism in humans. Metabolism. 2012, 61: 1495-1511. 10.1016/j.metabol.2012.07.007.CrossRefPubMed
18.
Oka M, Maeda S, Koga N, Kato K, Saito T: A modified colorimetric MTT assay adapted for primary cultured hepatocytes: application to proliferation and cytotoxicity assays. Biosci Biotechnol Biochem. 1992, 56: 1472-1473. 10.1271/bbb.56.1472.CrossRefPubMed
19.
Li Volti G, Wang J, Traganos F, Kappas A, Abraham NG: Differential effect of heme oxygenase-1 in endothelial and smooth muscle cell cycle progression. Biochem Biophys Res Commun. 2002, 296: 1077-1082. 10.1016/S0006-291X(02)02054-5.CrossRefPubMed
20.
Chastre A, Jiang W, Desjardins P, Butterworth RF: Ammonia and proinflammatory cytokines modify expression of genes coding for astrocytic proteins implicated in brain edema in acute liver failure. Metab Brain Dis. 2010, 25: 17-21. 10.1007/s11011-010-9185-y.CrossRefPubMed
21.
Jayakumar AR, Panickar KS, Murthy CR, Norenberg MD: Oxidative stress and mitogen-activated protein kinase phosphorylation mediate ammonia-induced cell swelling and glutamate uptake inhibition in cultured astrocytes. J Neurosci. 2006, 26: 4774-4784. 10.1523/JNEUROSCI.0120-06.2006.CrossRefPubMed
22.
23.
Hassell T, Glevae S, Butler M: Growth inhibition in animal cell culture. Appl Biochem Biotechnol. 1991, 30: 29-41. 10.1007/BF02922022.CrossRefPubMed
24.
Brusilow SW, Koehler RC, Traystman RJ, Cooper AJ: Astrocyte glutamine synthetase: importance in hyperammonemic syndromes and potential target for therapy. Neurotherapeutics. 2010, 7: 452-470. 10.1016/j.nurt.2010.05.015.CrossRefPubMedPubMedCentral
25.
Garcovich M, Zocco MA, Roccarina D, Ponziani FR, Gasbarrini A: Prevention and treatment of hepatic encephalopathy: focusing on gut microbiota. World J Gastroenterol. 2012, 18: 6693-6700. 10.3748/wjg.v18.i46.6693.CrossRefPubMedPubMedCentral
26.
Jones EA, Mullen KD: Theories of the pathogenesis of hepatic encephalopathy. Clin Liver Dis. 2012, 16: 7-26. 10.1016/j.cld.2011.12.010.CrossRefPubMed
27.
Atluri DK, Prakash R, Mullen KD: Pathogenesis, diagnosis, and treatment of hepatic encephalopathy. J Clin Exp Hepatol. 2011, 1: 77-86. 10.1016/S0973-6883(11)60126-6.CrossRefPubMedPubMedCentral
28.
Kala G, Kumarathasan R, Peng L, Leenen FH, Hertz L: Stimulation of Na+, K + -ATPase activity, increase in potassium uptake, and enhanced production of ouabain-like compounds in ammonia-treated mouse astrocytes. Neurochem Int. 2000, 36: 203-211. 10.1016/S0197-0186(99)00117-5.CrossRefPubMed
29.
Xue Z, Li B, Gu L, Hu X, Li M, Butterworth RF, Peng L: Increased Na, K-ATPase alpha2 isoform gene expression by ammonia in astrocytes and in brain in vivo. Neurochem Int. 2010, 57: 395-403. 10.1016/j.neuint.2010.04.014.CrossRefPubMed
30.
Jia B, Zujiang Y, Zhenfeng D, Lü XQ, Li JJ, Liu XR, Sun R, Gao XJ, Wang YF, Yan JY, Kan QC: Hyperammonaemia induces hepatic injury with alteration of gene expression profiles. Liver Int. 2014, 34 (5): 748-758. 10.1111/liv.12365.CrossRefPubMed
31.
BERNARDI P, FORTE M: The mitochondrial permeability transition pore. Novartis Found Symp. 2007, 287: 157-164. discussion 64-9CrossRefPubMed
32.
33.
Julie NL, Julie IM, Kende AI, Wilson GL: Mitochondrial dysfunction and delayed hepatotoxicity: another lesson from troglitazone. Diabetologia. 2008, 51: 2108-2116. 10.1007/s00125-008-1133-6.CrossRefPubMed
34.
Kessova IG, Cederbaum AI: Mitochondrial alterations in livers of Sod1-/- mice fed alcohol. Free Radic Biol Med. 2007, 42: 1470-1480. 10.1016/j.freeradbiomed.2007.01.044.CrossRefPubMedPubMedCentral
35.
Kim JH, Choi S, Jung JE, Roh EJ, Kim HJ: Capacitative Ca2+ entry is involved in regulating soluble amyloid precursor protein (sAPPalpha) release mediated by muscarinic acetylcholine receptor activation in neuroblastoma SH-SY5Y cells. J Neurochem. 2006, 97: 245-254. 10.1111/j.1471-4159.2006.03734.x.CrossRefPubMed
36.
Jang YJ, Ryu HJ, Choi YO, Kim C, Leem CH, Park CS: Improvement of insulin sensitivity by chelation of intracellular Ca(2+) in high-fat-fed rats. Metabolism. 2002, 51: 912-918. 10.1053/meta.2002.33351.CrossRefPubMed
37.
38.
Lowenstein CJ, Dinerman JL, Snyder SH: Nitric oxide: a physiologic messenger. Ann Intern Med. 1994, 120: 227-237. 10.7326/0003-4819-120-3-199402010-00009.CrossRefPubMed
39.
Yamaguchi M: The anti-apoptotic effect of regucalcin is mediated through multisignaling pathways. Apoptosis. 2013, 18: 1145-1153. 10.1007/s10495-013-0859-x.CrossRefPubMedPubMedCentral
40.
Huttemann M, Pecina P, Rainbolt M, Sanderson TH, Kagan VE, Samavati L, Doan JW, Lee I: The multiple functions of cytochrome c and their regulation in life and death decisions of the mammalian cell: From respiration to apoptosis. Mitochondrion. 2011, 11: 369-381. 10.1016/j.mito.2011.01.010.CrossRefPubMedPubMedCentral
41.
Wang X: The expanding role of mitochondria in apoptosis. Genes Dev. 2001, 15: 2922-2933.PubMed
42.
Green DR: Apoptotic pathways: paper wraps stone blunts scissors. Cell. 2000, 102: 1-4. 10.1016/S0092-8674(00)00003-9.CrossRefPubMed
43.
Li K, Li Y, Shelton JM, Richardson JA, Spencer E, Chen ZJ, Wang X, Williams RS: Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell. 2000, 101: 389-399. 10.1016/S0092-8674(00)80849-1.CrossRefPubMed
Metadata
Title
Hyperammonia induces specific liver injury through an intrinsic Ca2+-independent apoptosis pathway
Authors
Jingjing Li
Zujiang Yu
Qiongye Wang
Duolu Li
Bin Jia
Yubing Zhou
Yanwei Ye
Shen Shen
Yanfang Wang
Shasha Li
Lu Bai
Quancheng Kan
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Gastroenterology / Issue 1/2014
Electronic ISSN: 1471-230X
DOI
https://doi.org/10.1186/1471-230X-14-151

Other articles of this Issue 1/2014

BMC Gastroenterology 1/2014 Go to the issue

Case report

Lymphomatoid granulomatosis associated with azathioprine therapy in Crohn disease

Case report

Intestinal obstruction due to dual gastrointestinal atresia in infants: diagnosis and management of 3 cases

Case report

Severe cholestatic jaundice after a single administration of ajmaline; a case report and review of the literature

Research article

NOX2-generated oxidative stress is associated with severity of ultrasound liver steatosis in patients with non-alcoholic fatty liver disease

Research article

Fluorescence imaging in vivo visualizes delayed gastric emptying of liquid enteral nutrition containing pectin

Research article

Trends in gastroenteritis-associated mortality in the United States, 1985–2005: variations by ICD-9 and ICD-10 codes

ACC 2024 Congress Coverage

Cardiology Video

Highlights from the ACC 2024 Congress

Watch now

Watch official videos from the ACC congress summarizing key highlights from the last year in heart failure, cardiomyopathies, valvular disease, pulmonary vascular disease, and pediatric cardiology.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits