Attenuating effects of allantoin on oxidative stress in a mouse model of nonalcoholic steatohepatitis: role of SIRT1/Nrf2 pathway

Zeinab Hamidi-zad , Azam Moslehi , Maryam Rastegarpanah


Background and purpose: Nonalcoholic steatohepatitis (NASH) is considered a common and serious liver disease, which develops into cirrhosis, fibrosis, and even hepatocellular carcinoma. Oxidative stress is identified as an important factor in the induction and promotion of NASH. Allantoin is a natural and safe compound and has notable effects on lipid metabolism, inflammation, and oxidative stress. Therefore, this study was aimed to assess the role of allantoin on the oxidative stress and SIRT1/Nrf2 pathway in a mouse model of NASH.

Experimental approach: C57/BL6 male mice received saline and allantoin (saline as the control and allantoin as the positive control groups). NASH was induced by a methionine-choline deficient diet (MCD). In the NASH-allantoin (NASH-Alla) group, allantoin was injected for 4 weeks in the mice feeding on an MCD diet. Afterward, histopathological, serum, oxidative stress, and western blot evaluations were performed.

Findings/Results: We found NASH provided hepatic lipid accumulation and inflammation. Superoxide dismutase (SOD) and glutathione (GSH) levels decreased, lipid peroxidation increased, and the expression of SIRT1 and Nrf2 downregulated. However, allantoin-treatment decreased serum cholesterol, ALT, and AST. Liver steatosis and inflammation were improved. Protein expression of SIRT1 and Nrf2 were upregulated and SOD, CAT, and GSH levels increased and lipid peroxidation decreased.

Conclusion and implications: It seems that the antioxidant effects of allantoin might have resulted from the activation of SIRT1/Nrf2 pathway and increase of cellular antioxidant power.




Keywords: Allantoin; NASH; Nrf2; Oxidative stress; SIRT1.

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Gottlieb A, Mosthael W, Sowa JP, Canbay A. Nonalcoholic-fatty-liver-disease and nonalcoholic steatohepatitis: successful development of pharmacological treatment will depend on translational research. Digestion. 2019;100(2):79-85.

DOI: 10.1159/000493259.

Wree A, Broderick L, Canbay A, Hoffman HM, Feldstein AE. From NAFLD to NASH to cirrhosis-new insights into disease mechanisms. Nat Rev Gastroenterol Hepatol. 2013;10(11):627-636.

DOI: 10.1038/nrgastro.2013.149.

El-Lakkany NM, Seif El-Din SH, Sabra AA, Hammam OA, Ebeid FA. Co-administration of metformin and N-acetylcysteine with dietary control improves the biochemical and histological manifestations in rats with non-alcoholic fatty liver. Res Pharm Sci. 2016;11(5):374-382.

DOI: 10.4103/1735-5362.192487.

Liu W, Baker RD, Bhatia T, Zhu L, Baker SS. Pathogenesis of nonalcoholic steatohepatitis. Cell Mol Liff Sci. 2016;73(10):1969-1987.

DOI: 10.1007/s00018-016-2161-x.

Nili-Ahmadabadi A, Akbari Z, Ahmadimoghaddam D, Larki-Harchegani A. The role of ghrelin and tumor necrosis factor alpha in diazinon-induced dyslipidemia: insights into energy balance regulation. Pestic Biochem Physiol. 2019;157:138-142.

DOI: 10.1016/j.pestbp.2019.03.013.

Ibrahim SH, Hirsova P, Gores GJ. Non-alcoholic steatohepatitis pathogenesis: sublethal hepatocyte injury as a driver of liver inflammation. Gut. 2018;67(5):963-972.

DOI: 10.1136/gutjnl-2017-315691.

Nili-Ahmadabadi A, Alibolandi P, Ranjbar A, Mousavi L, Nili-Ahmadabadi H, Larki-Harchegani A, et al. Thymoquinone attenuates hepatotoxicity and oxidative damage caused by diazinon: an in vivo study. Res Pharm Sci. 2018;13(6):500-508.

DOI: 10.4103/1735-5362.245962.

Takaki A, Kawai D, Yamamoto K. Molecular mechanisms and new treatment strategies for non-alcoholic steatohepatitis (NASH). Int J Mol Sci. 2014;15(5):7352-7379.

DOI: 10.3390/ijms15057352.

Singh V, Ubaid S. Role of silent information regulator 1 (SIRT1) in regulating oxidative stress and inflammation. Inflammation. 2020;43(5):1589-1598.

DOI: 10.1007/s10753-020-01242-9.

Rodgers JT, Puigserver P. Fasting-dependent glucose and lipid metabolic response through hepatic sirtuin 1. Proc Natl Acad Sci U S A. 2007;104(31):12861-12866.

DOI: 10.1073/pnas.0702509104.

Samimi F, Baazm M, Eftekhar E, Rajabi S, Goodarzi MT, Jalali Mashayekhi F. Possible antioxidant mechanism of coenzyme Q10 in diabetes: impact on Sirt1/Nrf2 signaling pathways. Res Pharm Sci. 2019;14(6):524-533.

DOI: 10.4103/1735-5362.272561.

Xin FZ, Zhao ZH, Zhang RN, Pan Q, Gong ZZ, Sun C, et al. Folic acid attenuates high-fat diet-induced steatohepatitis via deacetylase SIRT1-dependent restoration of PPARα. World J Gastroenterol. 2020;26(18):2203-2220.

DOI: 10.3748/wjg.v26.i18.2203.

Jin SH, Yang JH, Shin BY, Seo K, Shin SM, Cho IJ, et al. Resveratrol inhibits LXRα-dependent hepatic lipogenesis through novel antioxidant Sestrin2 gene induction. Toxicol Appl Pharmacol. 2013;271(1):95-105.

DOI: 10.1016/j.taap.2013.04.023.

Chen MF, Yang TT, Yeh LR, Chung HH, Wen YJ, Lee WJ, et al. Activation of imidazoline I-2B receptors by allantoin to increase glucose uptake into C2C12 cells. Horm Metab Res. 2012;44(4):268-272.

DOI: 10.1055/s-0032-1301898.

Chen MF, Tsai JT, Chen LJ, Wu TP, Yang JJ, Yin LT, et al. Antihypertensive action of allantoin in animals. Biomed Res Int. 2014;2014:690135,1-6.

DOI: 10.1155/2014/690135.

Araújo LU, Grabe-Guimarães A, Mosqueira VC, Carneiro CM, Silva-Barcellos NM. Profile of wound healing process induced by allantoin. Acta Cir Bras. 2010;25(5):460-466.

DOI: 10.1590/s0102-86502010000500014.

Lin KC, Yeh LR, Chen LJ, Wen YJ, Cheng KC, Cheng JT. Plasma glucose-lowering action of allantoin is induced by activation of imidazoline I-2 receptors in streptozotocin-induced diabetic rats. Horm Metab Res. 2012;44(1):41-46.

DOI: 10.1055/s-0031-1295439.

da Silva DM, Martins JLR, de Oliveira DR, Florentino IF, da Silva DPB, Dos Santos FCA. Effect of allantoin on experimentally induced gastric ulcers: pathways of gastroprotection. Eur J Pharmacol. 2018;821:68-78.

DOI: 10.1016/j.ejphar.2017.12.052.

Ma J, Kang SY, Meng X, Kang AN, Park JH, Park YK, et al. Effects of rhizome extract of Dioscorea batatas and its active compound, allantoin, on the regulation of myoblast differentiation and mitochondrial biogenesis in C2C12 myotubes. Molecules. 2018;23(8):2023-2037.

DOI: 10.3390/molecules23082023.

Amitani M, Cheng KC, Asakawa A, Amitani H, Kairupan TS, Sameshima N, et al. Allantoin ameliorates chemically-induced pancreatic β-cell damage through activation of the imidazoline I3 receptors. PeerJ. 2015;3:e1105,1-15.

DOI: 10.7717/peerj.1105.

Komeili Movahhed T, Moslehi A, Golchoob M, Ababzadeh S. Allantoin improves methionine-choline deficient diet-induced nonalcoholic steatohepatitis in mice through involvement in endoplasmic reticulum stress and hepatocytes apoptosis-related genes expressions. Iran J Basic Med Sci. 2019;22(7):736-744.

DOI: 10.22038/ijbms.2019.33553.8012.

Yang TT, Chiu NH, Chung HH, Hsu CT, Lee WJ, Cheng JT. Stimulatory effect of allantoin on imidazoline I1 receptors in animal and cell line. Horm Metab Res. 2012;44(12):879-884.

DOI: 10.1055/s-0032-1312624.

Nikoukar LR, Nabavizadeh F, Mohamadi SM, Moslehi A, Hassanzadeh G, Nahrevanian H, et al. Protective effect of ghrelin in a rat model of celiac disease. Acta Physiol Hung. 2014;101(4):438-447.

DOI: 10.1556/APhysiol.101.2014.4.5.

Machado MV, Michelotti GA, Xie G, Almeida Pereira T, Boursier J, Bohnic B, et al. Mouse models of diet-induced nonalcoholic steatohepatitis reproduce the heterogeneity of the human disease. PLoS One. 2015;10(5):e0127991,1-16.

DOI: 10.1371/journal.pone.0127991.

Kleiner DE, Brunt EM, Van Natta M, Behling C, Contos MJ, Cummings OW, et al. Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology. 2005;41(6):1313-1321.

DOI: 10.1002/hep.20701.

Lee MY, Lee NH, Jung D, Lee JA, Seo CS, Lee H, et al. Protective effects of allantoin against ovalbumin (OVA)-induced lung inflammation in a murine model of asthma. Int Immunopharmacol. 2010;10(4):474-480.

DOI: 10.1016/j.intimp.2010.01.008.

Yang R, Song C, Chen J, Zhou L, Jiang X, Cao X, et al. Limonin ameliorates acetaminophen-induced hepatotoxicity by activating Nrf2 antioxidative pathway and inhibiting NF-κB inflammatory response via upregulating Sirt1. Phytomedicine. 2020;69:153211.

DOI: 10.1016/j.phymed.2020.153211.

Min HK, Kapoor A, Fuchs M, Mirshahi F, Zhou H, Maher J, et al. Increased hepatic synthesis and dysregulation of cholesterol metabolism is associated with the severity of nonalcoholic fatty liver disease. Cell Metab. 2012;15(5):665-674.

DOI: 10.1016/j.cmet.2012.04.004.

Yan T, Huang J, Nisar MF, Wan C, Huang W. The beneficial roles of SIRT1 in drug-induced liver injury. Oxid Med Cell Longev. 2019;2019:8506195,1-14.

DOI: 10.1155/2019/8506195.

Tang W, Jiang YF, Ponnusamy M, Diallo M. Role of Nrf2 in chronic liver disease. World J Gastroenterol. 2014;20(36):13079-13087.

DOI: 10.3748/wjg.v20.i36.13079.

Huang K, Huang J, Xie X, Wang S, Chen C, Shen X, et al. Sirt1 resists advanced glycation end products-induced expressions of fibronectin and TGF-β1 by activating the Nrf2/ARE pathway in glomerular mesangial cells. Free Radic Biol Med. 2013;65:528-540.

DOI: 10.1016/j.freeradbiomed.2013.07.029.

Zhong X, Liu H. Baicalin attenuates diet induced nonalcoholic steatohepatitis by inhibiting inflammation and oxidative stress via suppressing JNK signaling pathways. Biomed Pharmacother. 2018;98:111-117.

DOI: 10.1016/j.biopha.2017.12.026.

Santos-López JA, Garcimartín A, Merino P, López-Oliva ME, Bastida S, Benedí J, et al. Effects of silicon vs. hydroxytyrosol-enriched restructured pork on liver oxidation status of aged rats fed high-saturated/high-cholesterol diets. PLoS One. 2016;11(1):e0147469,1-16.

DOI: 10.1371/journal.pone.0147469.

Ma J, Meng X, Liu Y, Yin C, Zhang T, Wang P, et al. Effects of a rhizome aqueous extract of Dioscorea batatas and its bioactive compound, allantoin in high fat diet and streptozotocin-induced diabetic mice and the regulation of liver, pancreas and skeletal muscle dysfunction. J Ethnopharmacol. 2020;259:112926,1-10.

DOI: 10.1016/j.jep.2020.112926.

Eslami-Farsani M, Moslehi A, Hatami-Shahmir A. Allantoin improves histopathological evaluations in a rat model of gastritis. Physiol Int. 2018;105(4):325-334.

DOI: 10.1556/2060.105.2018.4.30.


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