Protective effects of compound ammonium glycyrrhizin, L‑arginine, silymarin and glucurolactone against liver damage induced by ochratoxin A in primary chicken hepatocytes

Yu,Zugong; Wu,Feng; Tian,Jing; Guo,Xuewen; An,Ran

doi:10.3892/mmr.2018.9285

Protective effects of compound ammonium glycyrrhizin, L‑arginine, silymarin and glucurolactone against liver damage induced by ochratoxin A in primary chicken hepatocytes

Authors:
- Zugong Yu
- Feng Wu
- Jing Tian
- Xuewen Guo
- Ran An
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Affiliations: Laboratory of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China
Published online on: July 16, 2018 https://doi.org/10.3892/mmr.2018.9285
Pages: 2551-2560
Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ochratoxin A (OTA) is a mycotoxin that is produced by fungi in improperly stored food and animal feed. It exhibits nephrotoxic, hepatotoxic, embryotoxic, teratogenic, neurotoxic, immunotoxic and carcinogenic effects in laboratory and farm animals. In the present study, the hepatotoxicity of OPA was investigated in chicken primary hepatocytes. On this basis, the cytoprotective effects of compound ammonium glycyrrhizin (CAG), L‑arginine (L‑Arg), silymarin (Sil) and glucurolactone (GA) were investigated in vitro. Hepatocytes were treated with OTA, which resulted in a significant decrease in cell viability and increases in serum aspartate transaminase and alanine transaminase activities, as determined by an MTT assay and commercial kits, respectively. Furthermore, following OTA treatment, the levels of hepatic antioxidants, such as superoxide dismutase and glutathione, were decreased, and the lipid peroxidation product malondialdehyde was increased, compared with the control group. However, pretreatment with CAG, L‑Arg, Sil and GA significantly ameliorated these alterations and Sil exerted the optimum hepatoprotective effect. The apoptotic rates were measured by flow cytometry and the results revealed that OTA increased cell apoptosis. The four types of hepatoprotective compounds employed in the present study decreased the apoptosis rate and significantly reversed OTA‑induced increases in the mRNA expression levels of caspase‑3, which was determined by reverse transcription‑quantitative polymerase chain reaction. Furthermore, B‑cell lymphoma‑2 (Bcl‑2) mRNA expression was increased in OTA‑treated cells when pretreated with CAG, L‑Arg, Sil and GA. However, no alterations in the mRNA expression of Bcl‑2‑associated X were observed in the L‑Arg and GA groups, compared with the OTA‑only group. These results indicate that OTA may exhibit hepatotoxicity in chickens and that CAG, L‑Arg, Sil and GA may protect the liver against this via anti‑oxidative and antiapoptosis mechanisms. In addition, CAG and GA are likely to mediate their effects through the mitochondrion‑dependent apoptosis pathway; however, the exact hepatoprotective mechanism of L‑Arg and GA require further investigation. Therefore, CAG, L‑Arg, Sil and GA are potential candidates for the prevention and treatment of chicken liver injury.

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1	Coronel MB, Sanchis V, Ramos AJ and Marin S: Review. Ochratoxin A: Presence in human plasma and intake estimation. Food Sci Technol Int. 16:5–18. 2010. View Article : Google Scholar : PubMed/NCBI
2	O'Brien E and Dietrich DR: Ochratoxin A: The continuing enigma. Crit Rev Toxicol. 35:33–60. 2005. View Article : Google Scholar : PubMed/NCBI
3	Ringot D, Chango A, Schneider YJ and Larondelle Y: Toxicokinetics and toxicodynamics of ochratoxin A, an update. Chem Biol Interact. 159:18–46. 2006. View Article : Google Scholar : PubMed/NCBI
4	Skaug MA, Helland I, Solvoll K and Saugstad OD: Presence of ochratoxin A in human milk in relation to dietary intake. Food Addit Contam. 18:321–327. 2001. View Article : Google Scholar : PubMed/NCBI
5	International Agency for Research on Cancer: Some naturally occurring substances: Food items and constituents, heterocyclic aromatic amines and mycotoxins, . Apresentado em: IARC Working Group on the Evaluation of Carcinogenic Risks to Humans: Some Naturally Occurring Substances: Food Items and Constituents. Lyon: 1992
6	Malir F, Ostry V, Pfohl-Leszkowicz A and Novotna E: Ochratoxin A: Developmental and reproductive toxicity-an overview. Birth Defects Res B Dev Reprod Toxicol. 98:493–502. 2013. View Article : Google Scholar : PubMed/NCBI
7	Dopp E, Müller J, Hahnel C and Schiffmann D: Induction of genotoxic effects and modulation of the intracellular calcium level in syrian hamster embryo (SHE) fibroblasts caused by ochratoxin A. Food Chem Toxicol. 37:713–721. 1999. View Article : Google Scholar : PubMed/NCBI
8	Eder S, Benesic A, Freudinger R, Engert J, Schwerdt G, Drumm K and Gekle M: Nephritogenic ochratoxin A interferes with mitochondrial function and pH homeostasis in immortalized human kidney epithelial cells. Pflugers Arch. 440:521–529. 2000. View Article : Google Scholar : PubMed/NCBI
9	Schaaf GJ, Nijmeijer SM, Maas RF, Roestenberg P, de Groene EM and Fink-Gremmels J: The role of oxidative stress in the ochratoxin A-mediated toxicity in proximal tubular cells. Biochim Biophys Acta. 1588:149–158. 2002. View Article : Google Scholar : PubMed/NCBI
10	Ehrlich V, Darroudi F, Uhl M, Steinkellner H, Gann M, Majer BJ, Eisenbauer M and Knasmüller S: Genotoxic effects of ochratoxin A in human-derived hepatoma (HepG2) cells. Food Chem Toxicol. 40:1085–1090. 2002. View Article : Google Scholar : PubMed/NCBI
11	Hundhausen C, Bösch-Saadatmandi C, Augustin K, Blank R, Wolffram S and Rimbach G: Effect of vitamin E and polyphenols on ochratoxin A-induced cytotoxicity in liver (HepG2) cells. J Plant Physiol. 162:818–822. 2005. View Article : Google Scholar : PubMed/NCBI
12	Arbillaga L, Azqueta A, Ezpeleta O and de Cerain López A: Oxidative DNA damage induced by ochratoxin A in the HK-2 human kidney cell line: Evidence of the relationship with cytotoxicity. Mutagenesis. 22:35–42. 2006. View Article : Google Scholar : PubMed/NCBI
13	Guerra M, Galvano F, Bonsi L, Speroni E, Costa S, Renzulli C and Cervellati R: Cyanidin-3-O-beta-glucopyranoside, a natural free-radical scavenger against aflatoxin B1-and ochratoxin A-induced cell damage in a human hepatoma cell line (Hep G2) and a human colonic adenocarcinoma cell line (CaCo-2). Br J Nutr. 94:211–220. 2005. View Article : Google Scholar : PubMed/NCBI
14	Kamp HG, Eisenbrand G, Schlatter J, Würth K and Janzowski C: Ochratoxin A: Induction of (oxidative) DNA damage, cytotoxicity and apoptosis in mammalian cell lines and primary cells. Toxicology. 206:413–425. 2005. View Article : Google Scholar : PubMed/NCBI
15	Li J, Yin S, Dong Y, Fan L and Hu H: p53 activation inhibits ochratoxin A-induced apoptosis in monkey and human kidney epithelial cells via suppression of JNK activation. Biochem Biophys Res Commun. 411:458–463. 2011. View Article : Google Scholar : PubMed/NCBI
16	Li JY, Cao HY, Liu P, Cheng GH and Sun MY: Glycyrrhizic acid in the treatment of liver diseases: Literature review. Biomed Res Int. 2014:8721392014.PubMed/NCBI
17	Arase Y, Ikeda K, Murashima N, Chayama K, Tsubota A, Koida I, Suzuki Y, Saitoh S, Kobayashi M and Kumada H: The long term efficacy of glycyrrhizin in chronic hepatitis C patients. Cancer. 79:1494–1500. 1997. View Article : Google Scholar : PubMed/NCBI
18	Nishimoto Y, Hisatsune A, Katsuki H, Miyata T, Yokomizo K and Isohama Y: Glycyrrhizin attenuates mucus production by inhibition of MUC5AC mRNA expression in vivo and in vitro. J Pharmacol Sci. 113:76–83. 2010. View Article : Google Scholar : PubMed/NCBI
19	Visavadiya NP and Narasimhacharya AV: Hypocholesterolaemic and antioxidant effects of Glycyrrhiza glabra (Linn) in rats. Mol Nutr Food Res. 50:1080–1086. 2006. View Article : Google Scholar : PubMed/NCBI
20	Oh HM, Lee S, Park YN, Choi EJ, Choi JY, Kim JA, Kweon JH, Han WC, Choi SC, Han JK, et al: Ammonium glycyrrhizinate protects gastric epithelial cells from hydrogen peroxide-induced cell death. Exp Biol Med (Maywood). 234:263–277. 2009. View Article : Google Scholar : PubMed/NCBI
21	Račková L, Jančinová V, Petríková M, Drábiková K, Nosál R, Stefek M, Kostálová D, Prónayová N and Kovácová M: Mechanism of anti-inflammatory action of liquorice extract and glycyrrhizin. Nat Prod Res. 21:1234–1241. 2007. View Article : Google Scholar : PubMed/NCBI
22	Yim SB, Park SE and Lee CS: Protective effect of glycyrrhizin on 1-methyl-4-phenylpyridinium-induced mitochondrial damage and cell death in differentiated PC12 cells. J Pharmacol Exp Ther. 321:816–822. 2007. View Article : Google Scholar : PubMed/NCBI
23	Lass A, Suessenbacher A, Wölkart G, Mayer B and Brunner F: Functional and analytical evidence for scavenging of oxygen radicals by L-arginine. Mol Pharmacol. 61:1081–1088. 2002. View Article : Google Scholar : PubMed/NCBI
24	Wu G, Bazer FW, Davis TA, Kim SW, Li P, Rhoads Marc J, Satterfield Carey M, Smith SB, Spencer TE and Yin Y: Arginine metabolism and nutrition in growth, health and disease. Amino Acids. 37:153–168. 2009. View Article : Google Scholar : PubMed/NCBI
25	Wu G, Davis PK, Flynn NE, Knabe DA and Davidson JT: Endogenous synthesis of arginine plays an important role in maintaining arginine homeostasis in postweaning growing pigs. J Nutr. 127:2342–2349. 1997. View Article : Google Scholar : PubMed/NCBI
26	Chen CH, Huang TS, Wong CH, Hong CL, Tsai YH, Liang CC, Lu FJ and Chang WH: Synergistic anti-cancer effect of baicalein and silymarin on human hepatoma HepG2 cells. Food Chem Toxicol. 47:638–644. 2009. View Article : Google Scholar : PubMed/NCBI
27	Jose MA, Abraham A and Narmadha M: Effect of silymarin in diabetes mellitus patients with liver diseases. J Pharmacol Pharmacother. 2:287–289. 2011. View Article : Google Scholar : PubMed/NCBI
28	Guan S, Gong M, Yin Y, Huang R, Ruan Z, Zhou T and Xie M: Occurrence of mycotoxins in feeds and feed ingredients in China. J Food Agric Environ. 9:163–167. 2011.
29	Yu Z, Wu F, Tian J, Guo X, An R and Guo Y: Ammonium glycyrrhizin counteracts liver injury caused by lipopolysaccharide/amoxicillin-clavulanate potassium. Oncotarget. 8:96837–96851. 2017.PubMed/NCBI
30	Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
31	Schefe JH, Lehmann KE, Buschmann IR, Unger T and Funke-Kaiser H: Quantitative real-time RT-PCR data analysis: Current concepts and the novel ‘gene expression's C T difference’ formula. J Mol Med (Berl). 84:901–910. 2006. View Article : Google Scholar : PubMed/NCBI
32	Petzinger E and Ziegler K: Ochratoxin A from a toxicological perspective. J Vet Pharmacol Ther. 23:91–98. 2000. View Article : Google Scholar : PubMed/NCBI
33	Li Y, Zhang B, He X, Cheng WH, Xu W, Luo Y, Liang R, Luo H and Huang K: Analysis of individual and combined effects of ochratoxin A and zearalenone on HepG2 and KK-1 cells with mathematical models. Toxins (Basel). 6:1177–1192. 2014. View Article : Google Scholar : PubMed/NCBI
34	Li Y, He X, Yang X, Huang K, Luo Y, Zhu L, Li Y and Xu W: Zinc inhibits the reproductive toxicity of Zearalenone in immortalized murine ovarian granular KK-1 cells. Sci Rep. 5:142772015. View Article : Google Scholar : PubMed/NCBI
35	Klarić MŠ, Želježić D, Rumora L, Peraica M, Pepeljnjak S and Domijan AM: A potential role of calcium in apoptosis and aberrant chromatin forms in porcine kidney PK15 cells induced by individual and combined ochratoxin A and citrinin. Arch Toxicol. 86:97–107. 2012. View Article : Google Scholar : PubMed/NCBI
36	Wilk-Zasadna I and Minta M: Developmental toxicity of ochratoxin A in rat embryo midbrain micromass cultures. Int J Mol Sci. 10:37–49. 2008. View Article : Google Scholar : PubMed/NCBI
37	Hassen W, Ayed-Boussema I, Oscoz AA, Ade Lopez C and Bacha H: The role of oxidative stress in zearalenone-mediated toxicity in Hep G2 cells: Oxidative DNA damage, gluthatione depletion and stress proteins induction. Toxicology. 232:294–302. 2007. View Article : Google Scholar : PubMed/NCBI
38	Abid-Essefi S, Ouanes Z, Hassen W, Baudrimont I, Creppy E and Bacha H: Cytotoxicity, inhibition of DNA and protein syntheses and oxidative damage in cultured cells exposed to zearalenone. Toxicol In Vitro. 18:467–474. 2004. View Article : Google Scholar : PubMed/NCBI
39	Zheng J, Zhang Y, Xu W, Luo Y, Hao J, Shen XL, Yang X, Li X and Huang K: Zinc protects HepG2 cells against the oxidative damage and DNA damage induced by ochratoxin A. Toxicol Appl Pharmacol. 268:123–131. 2013. View Article : Google Scholar : PubMed/NCBI
40	Klarić MS, Pepeljnjak S, Domijan A-M and Petrik J: Lipid peroxidation and glutathione levels in porcine kidney PK15 cells after individual and combined treatment with fumonisin B1, beauvericin and ochratoxin A. Basic Clin Pharmacol Toxicol. 100:157–164. 2007. View Article : Google Scholar : PubMed/NCBI
41	Lawen A: Apoptosis-an introduction. Bioessays. 25:888–896. 2003. View Article : Google Scholar : PubMed/NCBI
42	Bouaziz C, el dein Sharaf O, Martel C, El Golli E, Abid-Essefi S, Brenner C, Lemaire C and Bacha H: Molecular events involved in ochratoxin A induced mitochondrial pathway of apoptosis, modulation by Bcl-2 family members. Environ Toxicol. 26:579–590. 2011. View Article : Google Scholar : PubMed/NCBI
43	El Golli Bennour E, Rodriguez-Enfedaque A, Bouaziz C, Ladjimi M, Renaud F and Bacha H: Toxicities induced in cultured human hepatocarcinoma cells exposed to ochratoxin A: Oxidative stress and apoptosis status. J Biochem Mol Toxicol. 23:87–96. 2009. View Article : Google Scholar : PubMed/NCBI
44	Ming LJ and Yin A: Therapeutic effects of glycyrrhizic acid. Nat Prod Commun. 8:415–418. 2013.PubMed/NCBI
45	Lee CH, Park SW, Kim YS, Kang SS, Kim JA, Lee SH and Lee SM: Protective mechanism of glycyrrhizin on acute liver injury induced by carbon tetrachloride in mice. Biol Pharm Bull. 30:1898–1904. 2007. View Article : Google Scholar : PubMed/NCBI
46	Van Rossum T, Vulto AG, de Man RA, Brouwer JT and Schalm SW: Review article: Glycyrrhizin as a potential treatment for chronic hepatitis C. Aliment Pharmacol Ther. 12:199–205. 1998. View Article : Google Scholar : PubMed/NCBI
47	Tapiero H, Mathé G, Couvreur P and Tew K: I. Arginine. Biomed Pharmacother. 56:439–445. 2002. View Article : Google Scholar : PubMed/NCBI
48	Andrew PJ and Mayer B: Enzymatic function of nitric oxide synthases. Cardiovasc Res. 43:521–531. 1999. View Article : Google Scholar : PubMed/NCBI
49	Soto C, Pérez J, García V, Uría E, Vadillo M and Raya L: Effect of silymarin on kidneys of rats suffering from alloxan-induced diabetes mellitus. Phytomedicine. 17:1090–1094. 2010. View Article : Google Scholar : PubMed/NCBI
50	Stanford D and Stachulski AV: Convenient syntheses of deoxypyranose sugars from glucuronolactone. Tetrahedron Lett. 48:2361–2364. 2007. View Article : Google Scholar
51	Yin G, Cao L, Xu P, Jeney G, Nakao M and Lu C: Hepatoprotective and antioxidant effects of Glycyrrhiza glabra extract against carbon tetrachloride (CCl(4))-induced hepatocyte damage in common carp (Cyprinus carpio). Fish Physiol Biochem. 37:209–216. 2011. View Article : Google Scholar : PubMed/NCBI
52	Dr S and Khanna A: The effect of antenatal L-arginine and antioxidant supplementation on oxidative stress marker levels in newborns. J Clin Diagn Res. 8:OC10–OC12. 2014.PubMed/NCBI
53	Kumar N, Rai A, Reddy ND, Raj PV, Jain P, Deshpande P, Mathew G, Kutty NG, Udupa N and Rao CM: Silymarin liposomes improves oral bioavailability of silybin besides targeting hepatocytes, and immune cells. Pharmacol Rep. 66:788–798. 2014. View Article : Google Scholar : PubMed/NCBI
54	El-Tahawy NF, Ali AH, Saied SR and Abdel-Wahab Z: Effect of glycyrrhizin on lipopolysaccharide/D-galactosamine-induced acute hepatitis in albino rats: A histological and immunohistochemical study. Egypt J Histol. 34:518–527. 2011. View Article : Google Scholar
55	Tuorkey MJ, El-Desouki NI and Kamel RA: Cytoprotective effect of silymarin against diabetes-induced cardiomyocyte apoptosis in diabetic rats. Biomed Environ Sci. 28:36–43. 2015.PubMed/NCBI