对乙酰氨基酚诱导的大鼠胆酸盐蓄积性肝损伤机制研究

doi:10.5246/jcps.2018.07.051

中国药学(英文版) ›› 2018, Vol. 27 ›› Issue (7): 498-509.DOI: 10.5246/jcps.2018.07.051

对乙酰氨基酚诱导的大鼠胆酸盐蓄积性肝损伤机制研究

靳永文¹, 饶志¹, 武艳芳^1,2, 张国强¹, 石阿茜¹, 魏玉辉¹, 武新安^1*

1. 兰州大学第一医院药剂科, 甘肃兰州 730000
2. 兰州大学药学院, 甘肃兰州 730000

收稿日期:2018-03-27 修回日期:2018-05-01 出版日期:2018-07-25 发布日期:2018-05-16
通讯作者: Tel.: +86-931-8616392, E-mail: xinanwu6511@163.com
基金资助:
The National Natural Science Foundation of China (Grant No. 81373494 and 81041086) and Tianqing Liver Disease Rearch Fund (Grant No. TQGB20180088).

Acetaminophen-induced hepatotoxicity in rats is correlated with the accumulation of bile acids: an underlying mechanism

Yongwen Jin¹, Zhi Rao¹, Yanfang Wu^1,2, Guoqiang Zhang¹, Axi Shi¹, Yuhui Wei¹, Xin’an Wu^1*

1. Department of Pharmacy, the First Hospital of Lanzhou University, Lanzhou 730000, China
2. College of Pharmaceutical Sience, Lanzhou University, Lanzhou 730000, China

Received:2018-03-27 Revised:2018-05-01 Online:2018-07-25 Published:2018-05-16
Contact: Tel.: +86-931-8616392, E-mail: xinanwu6511@163.com
Supported by:
The National Natural Science Foundation of China (Grant No. 81373494 and 81041086) and Tianqing Liver Disease Rearch Fund (Grant No. TQGB20180088).

摘要/Abstract

摘要：

通过测定血浆和肝组织中胆酸盐(BAs)含量的变化及肝组织中转运体表达水平的变化, 初步探究对乙酰氨基酚(APAP)致大鼠的肝损伤的机制。SD大鼠42只, 随机分为6-h对照组、APAP 6-h组、12-h对照组、APAP 12-h组、24-h对照组和APAP 24-h组。运用LC-MS/MS测定血浆和肝组织中BAs含量; Western blotting评价Bsep、Mrp2、Mrp4、Ntcp和Oatp2表达水平。APAP 6-h组, 与6-h对照组相比, 血浆和肝组织中BAs含量及肝组织中转运蛋白的表达水平无显著差异。APAP 12-h组, 与12-h对照组比较, 血浆中BAs含量明显降低(P<0.05), 肝组织中BAs含量明显升高(P<0.05); Bsep和Mrp2表达水平显著降低(P<0.05)。APAP 24-h组, 与24-h对照组比较, 血浆和肝组织中BAs含量均明显升高(P<0.05); Mrp4和Oatp2表达水平显着降低(P<0.05)。APAP诱导的大鼠肝损伤机制可能与其在不同时间抑制Bsep, Mrp2, Mrp4和Oatp2表达水平导致BAs蓄积有关。

关键词: 对乙酰氨基酚, 肝损伤, 胆酸盐, 肝组织转运体

Abstract:

In the present study, we aimed to investigate the underlying mechanism of acetaminophen (APAP)-induced hepatotoxicityby measuring the expression levels of liver transporters and concentrations of bile acids (BAs) in rat plasma and liver. SD rats (42)were randomly assigned into six groups, including 6-h control group, APAP 6-h group, 12-h control group, APAP 12-h group, 24-h control group and APAP 24-h group. The estimation study of BAs in plasma and liver was performed on LC-MS/MS.The levels of bile salt export pump (Bsep), multidrug resistant protein 2 (Mrp2), multidrug resistant protein 4 (Mrp4), Na⁺/taurocholate cotransporting polypeptide (Ntcp) and organic anion transporting polypeptide 2 (Oatp2) in the liver were analyzed by Western blotting analysis. Compared with the corresponding control groups, no difference was found in the BA levels and the expressions of BA transporters in the plasma and liver after 6 h of APAP administration. While BA levels were significantly decreased in the plasma and increased in the liver after 12 h of APAP administration (P<0.05); and the expressions of Bsep and Mrp2 were significantly reduced (P<0.05). After 24 h of APAP administration, BA levels were both greatly increased in the plasma and liver (P<0.05); and the expressions of Mrp4 and Oatp2 were significantly decreased (P<0.05). In response to over-dose APAP, Bsep, Mrp2, Mrp4 and Oatp2 levels were reduced at different time points, causing the accumulation of BAs, and such accumulation may ultimately lead to the severe liver injury, which could be an underlying mechanism of the APAP-induced hepatotoxicity.

Key words: Acetaminophen, Hepatotoxicity, Bile acids, Hepatobiliary transporter

中图分类号:

R963

Supporting:

靳永文, 饶志, 武艳芳, 张国强, 石阿茜, 魏玉辉, 武新安. 对乙酰氨基酚诱导的大鼠胆酸盐蓄积性肝损伤机制研究[J]. 中国药学(英文版), 2018, 27(7): 498-509.

Yongwen Jin, Zhi Rao, Yanfang Wu, Guoqiang Zhang, Axi Shi, Yuhui Wei, Xin’an Wu. Acetaminophen-induced hepatotoxicity in rats is correlated with the accumulation of bile acids: an underlying mechanism[J]. Journal of Chinese Pharmaceutical Sciences, 2018, 27(7): 498-509.

参考文献

[1] Danan, G.; Teschke, R. RUCAM in Drug and Herb Induced Liver Injury: The Update. Int. J. Mol. Sci. 2015, 17, 14.

[2] Shah, F.; Medvedev, A.; Wassermann, A.M.; Brodney, M.; Zhang, L.; Makarov, S.; Stanton, R.V. The Identification of Pivotal Transcriptional Factors Mediating Cell Responses to Drugs With Drug-Induced Liver Injury Liabilities. Toxicol. Sci. 2018, 162, 177-188.

[3] Schuster, D.; Laggner, C. Why drugs fail-a study on side effects in new chemical entities. Curr. Pharm. Des. 2005, 11, 3545-3559.

[4] Wagner, M.; Zollner, G. New molecular insights into the mechanisms of cholestasis. J. Hepatol. 2009, 51, 565-580.

[5] Anderson, N.; Borlak, J. Molecular mechanisms and therapeutic targets in steatosis and steatohepatitis. Toxicol. Sci. 2008, 60, 311-357.

[6] Jaeschke, H.; Gores, G.J. Mechanisms of hepatotoxicity. Toxicol. Sci. 2002, 65, 166-176.

[7] Jaeschke, H.; McGill, M.R. Oxidant stress, mitochondria, and cell death mechanisms in drug-induced liver injury: lessons learned from acetaminophen hepatotoxicity. Drug Metab. Rev. 2012, 44, 88-106.

[8] van Beusekom, C.D.; van den Heuvel, J.J. The feline bile salt export pump: a structural and functional comparison with canine and human Bsep/BSEP. BMC Vet. Res. 2013, 9, 259.

[9] Perez, M.J.; Briz, O. Bile-acid-induced cell injury and protection. World J. Gastroenterol. 2009, 15, 1677-1689.

[10] Beuers, U.; Hohenester, S. The biliary HCO(3)(-) umbrella: a unifying hypothesis on pathogenetic and therapeutic aspects of fibrosing cholangiopathies. Hepatology. 2010, 52, 1489-1496.

[11] Hohenester, S.; Wenniger, L.M. A biliary HCO3- umbrella constitutes a protective mechanism against bile acid-induced injury in human cholangiocytes. Hepatology. 2012, 55, 173-183.

[12] Shah, A.D.; Wood, D.M. Understanding lactic acidosis in paracetamol (acetaminophen) poisoning. Br. J. Clin. Pharmacol. 2011, 71, 20-28.

[13] Bunchorntavakul, C.; Reddy, K.R. Acetaminophen-related hepatotoxicity. Clin. Liver Dis. 2013, 17, 587-607.

[14] Ostapowicz, G.; Fontana, R.J. Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States. Ann. Intern. Med. 2002, 137, 947-954.

[15] Tan, S.C.; New, L.S. Prevention of acetaminophen (APAP)-induced hepatotoxicity by leflunomide via inhibition of APAP biotransformation to N-acetyl-p-benzoquinone imine. Toxicol. Lett. 2008, 180, 174-181.

[16] Dahlin, D.C.; Miwa, G.T. N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen. Proc. Natl. Acad. Sci. USA. 1984, 81, 1327-1331.

[17] Chen, W.; Koenigs, L.L. Oxidation of acetaminophen to its toxic quinone imine and nontoxic catechol metabolites by baculovirus-expressed and purified human cytochromes P450 2E1 and 2A6. Chem. Res. Toxicol. 1998, 11, 295-301.

[18] Aleksunes, L.M.; Slitt, A.L. Induction of Mrp3 and Mrp4 transporters during acetaminophen hepatotoxicity is dependent on Nrf2. Toxicol. Appl. Pharmacol. 2008, 226, 74-83.

[19] Ghanem, C.I.; Ruiz, M.L. Shift from biliary to urinary elimination of acetaminophen-glucuronide in aceta-minophen-pretreated rats. J. Pharmacol. Exp. Ther. 2005, 315, 987-995.
[20] Aleksunes, L.M.; Slitt, A.M. Differential expression of mouse hepatic transporter genes in response to aceta-minophen and carbon tetrachloride. Toxicol. Sci. 2005, 83, 44-52.

[21] Aleksunes, L.M.; Scheffer, G.L. Coordinated expression of multidrug resistance-associated proteins (Mrps) in mouse liver during toxicant-induced injury. Toxicol. Sci. 2006, 89, 370-379.

[22] Jin, Y.W.; Rao, Z. Development of a LC-MS/MS Method for Simultaneous Determination of Bile Acids and their Conjugates in Hepatocytes, Tissue and Fluids in Rat. Curr. Pharm. Anal. 2018, 14, 331-341.

[23] McJunkin, B.; Barwick, K.W. Fatal massive hepatic necrosis following acetaminophen overdose. JAMA. 1976, 236, 1874-1875.

[24] Larson, P.A.; Janower, M.L. The nighthawk: bird of paradise or albatross? J. Am. Coll. Radiol. 2005, 2, 967-970.

[25] Alkiyumi, S.S.; Abdullah, M.A. Ipomoea aquatica extract shows protective action against thioacetamide-induced hepatotoxicity. Molecules. 2012, 17, 6146-6155.

[26] James, A.P.; Slivkoff-Clark, K. Ipomoea aquatica extract shows protective action against thioacetamide-induced hepatotoxicity. Asia Pac. J. Public. Health. 2003, 15, 37-40.

[27] Jaeschke. H. Are cultured liver cells the right tool to investigate mechanisms of liver disease or hepatotoxicity? Hepatology. 2003, 38, 1053-1055.

[28] Hinson, J.P.; Khan, M. Dehydroepiandrosterone sulphate (DHEAS) inhibits growth of human vascular endothelial cells. Endocr. Res. 2004, 30, 667-671.

[29] Kon, T.; Tanigawa, T. Singlet oxygen quenching activity of human serum. Redox Rep. 2004, 9, 325-330.

[30] Chen, C.; Krausz, K.W. Serum metabolomics reveals irreversible inhibition of fatty acid beta-oxidation through the suppression of PPARalpha activation as a contributing mechanism of acetaminophen-induced hepatotoxicity. Chem. Res. Toxicol. 2009, 22, 699-707.

[31] Hylemon, P.B.; Zhou, H. Bile acids as regulatory molecules. J. Lipid Res. 2009, 50, 1509-1520.

[32] Perez, M.J.; Briz, O. Bile-acid-induced cell injury and protection. World J. Gastroenterology. 2009, 15, 1677-1689.

[33] Copple, B.L.; Jaeschke, H. Oxidative stress and the pathogenesis of cholestasis. Semin. Liver Dis. 2010, 30, 195-204.

[34] Luo. L.; Schomaker, S. Evaluation of serum bile acid profiles as biomarkers of liver injury in rodents. Toxicol. Sci. 2014, 137, 12-25.

[35] Alnouti, Y.; Csanaky, I.L. Quantitative-profiling of bile acids and their conjugates in mouse liver, bile, plasma, and urine using LC-MS/MS. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2008, 873, 209-217.

[36] Deo, A.K.; Bandiera, S.M. Biotransformation of lithocholic acid by rat hepatic microsomes: metabolite analysis by liquid chromatography/mass spectrometry. Drug Metab. Dispos. 2008, 36, 442-451.

[37] Rolo, A.P.; Oliveira, P.J. Bile acids affect liver mitochondrial bioenergetics: possible relevance for cholestasis therapy. Toxicol. Sci. 2000, 57, 177-185.

[38] Javitt, N.B.; Emerman, S. Effect of sodium taurolithocholate on bile flow and bile acid exeretion. J. Clin. Invest. 1968, 47, 1002.

[39] Fischer, C.; Cooper, N.; Rothschild, M.; Mosbach, E. Effect of dietary chenodeoxycholic acid and lithocholic acid in the rabbit. Dig. Dis. Sci. 1974, 19, 877-886.

[40] Delzenne, N.M.; Calderon, P.B. Comparative hepatotoxicity of cholic acid, deoxycholic acid and lithocholic acid in the rat: in vivo and in vitro studies. Toxicol. Lett. 1992, 61, 291-304.

[41] Palmer, A. Heywood, R. Pathological changes in the rhesus fetus associated with the oral administration of chenodeoxycholic acid. Toxicology. 1974, 2, 239-246.

[42] Dyrszka, H.; Salen, G. Pathological changes in the rhesus fetus associated with the oral administration of chenodeoxycholic acid. Gastroenterology. 1976, 70, 93.

[43] Song, P.; Zhang, Y. Dose-response of five bile acids on serum and liver bile Acid concentrations and hepatotoxicty in mice. Toxicol. Sci. 2011, 123, 359-367.

[44] Woolbright, B.L.; McGill, M.R. Acute Liver Failure Study Group. Glycodeoxycholic acid levels as prognostic biomarker in acetaminophen-induced acute liver failure patients. Toxicol. Sci. 2014, 142, 436-444.

[45] Rodrigues, C.M.; Stieers, C.L. Tauroursodeoxycholic acid partially prevents apoptosis induced by 3-nitropropionic acid: evidence for a mitochondrial pathway independent of the permeability transition. J. Neurochem. 2000, 75, 2368-2379.

[46] Trauner, M.; Boyer, J.L. Bile salt transporters: molecular characterization, function, and regulation. Physiol. Rev. 2003, 83, 633-671.

[47] Lorbek, G.; Lewinska, M. Cytochrome P450s in the synthesis of cholesterol and bile acids--from mouse models to human diseases. FEBS J. 2012, 279, 1516-1533.
[48] Trottier, J.; Białek, A. Profiling circulating and urinary bile acids in patients with biliary obstruction before and after biliary stenting. PLoS One. 2011, 6, e22094.

对乙酰氨基酚诱导的大鼠胆酸盐蓄积性肝损伤机制研究

Acetaminophen-induced hepatotoxicity in rats is correlated with the accumulation of bile acids: an underlying mechanism

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