, Volume 71, Issue 1, pp 181–192 | Cite as

Anti-hyperuricemic effect of isorhamnetin in cultured hepatocytes and model mice: structure–activity relationships of methylquercetins as inhibitors of uric acid production

  • Shin-ichi AdachiEmail author
  • Shinji Kondo
  • Yusuke Sato
  • Fumiaki Yoshizawa
  • Kazumi Yagasaki


Hyperuricemia is an important risk factor for gout. Isorhamnetin (3′-O-methylquercetin) is an O-methylated flavonol, which occurs in onion, almond and sea buckthorn. It is also one of the metabolites of quercetin in mammals. In the present study, we investigated anti-hyperuricemic effect of isorhamnetin adopting both cultured hepatocytes and mice with hyperuricemia induced by purine bodies. In cultured hepatocytes, isorhamnetin as well as quercetin significantly and dose-dependently inhibited uric acid (UA) production. We also examined the inhibitory effects on UA production of other mono-methylquercetins, i.e., tamarixetin, 3-O-methylquercetin, azaleatin, and rhamnetin in addition to isorhamnetin for studying their structure–activity relationships. From the results obtained, hydroxyl groups at C-3, C-5, and especially C-7, but not C-3′ and C-4′ of quercetin are demonstrated to play a critical role in suppressing UA production in the AML12 hepatocytes. Oral administration of isorhamnetin significantly reduced plasma and hepatic UA levels in the hyperuricemic model mice. Isorhamnetin also decreased hepatic xanthine oxidase (XO) activity without changes in XO protein expression, indicating that anti-hyperuricemic effect of isorhamnetin could be, at least partly, attributable to suppression of UA production by directly inhibiting XO activity in the liver. These findings demonstrate that isorhamnetin has a potent anti-hyperuricemic effect and may be a potential candidate for prevention and remediation of hyperuricemia.


Isorhamnetin AML12 hepatocyte Hyperuricemia Uric acid 



Balanced salt solution






Uric acid


Xanthine oxidase



This work was supported in part by the Reginal Innovation Strategy Support Program, MEXT, Japan, and in part by JSPS KAKENHI Grant Nos. JP16K16273 and JP15K07424. Authors are grateful to Keiichiro Numao, Kento Kobayashi, Kazusa Narita, Yuki Takami for their excellent technical assistance.

Compliance with ethical standards

Conflict of interest

Authors declare that they have no conflict of interest.


  1. Adachi S, Yoshizawa F, Yagasaki K (2017a) Assay systems for screening food and natural substances that have anti-hyperuricemic activity: uric acid production in cultured hepatocytes and purine bodies-induced hyperuricemic model mice. Cytotechnology 69:435–442. CrossRefGoogle Scholar
  2. Adachi S, Nihei K, Ishihara Y, Yoshizawa F, Yagasaki K (2017b) Anti-hyperuricemic effect of taxifolin in cultured hepatocytes and model mice. Cytotechnology 69:329–336. CrossRefGoogle Scholar
  3. Babio N, Martínez-González MA, Estruch R, Wärnberg J, Recondo J, Ortega-Calvo M, Serra-Majem L, Corella D, Fitó M, Ros E, Becerra-Tomás N, Basora J, Salas-Salvadó J (2015) Associations between serum uric acid concentrations and metabolic syndrome and its components in the PREDIMED study. Nutr Metab Cardiovasc Dis 25:173–180. CrossRefGoogle Scholar
  4. Chen C, Zhang H, Xiao W, Yong ZP, Bai N (2007) High-performance liquid chromatographic fingerprint analysis for different origins of sea buckthorn berries. J Chromatogr A 1154:250–259. CrossRefGoogle Scholar
  5. Chen G, Tan ML, Li KK, Leung PC, Ko CH (2015a) Green tea polyphenols decreases uric acid level through xanthine oxidase and renal urate transporters in hyperuricemic mice. J Ethnopharmacol 175:14–20. CrossRefGoogle Scholar
  6. Chen TL, Zhu GL, Wang JA, Zhang GD, Liu HF, Chen JR, Wang Y, He XL (2015b) Protective effects of isorhamnetin on apoptosis and inflammation in TNF-α-induced HUVECs injury. Int J Clin Exp Pathol 8:2311–2320Google Scholar
  7. Choi HK, Atkinson K, Karlson EW, Willett W, Curhan G (2004) Purine-rich foods, dairy and protein intake, and the risk of gout in men. N Engl J Med 350:1093–1103. CrossRefGoogle Scholar
  8. Cos P, Ying L, Calomme M, Hu JP, Cimanga K, Van Poel B, Pieters L, Vlietinck AJ, Vanden Berghe D (1998) Structure–activity relationship and classification of flavonoids as inhibitors of xanthine oxidase and superoxide scavengers. J Nat Prod 61:71–76. CrossRefGoogle Scholar
  9. Dong GZ, Lee JH, Ki SH, Yang JH, Cho IJ, Kang SH, Zhao RJ, Kim SC, Kim YW (2014) AMPK activation by isorhamnetin protects hepatocytes against oxidative stress and mitochondrial dysfunction. Eur J Pharmacol 740:634–640. CrossRefGoogle Scholar
  10. Edwards RL, Lyon T, Litwin SE, Rabovsky A, Symons JD, Jalili T (2007) Quercetin reduces blood pressure in hypertensive subjects. J Nutr 137:2405–2411. CrossRefGoogle Scholar
  11. Frampton JE (2015) Febuxostat: a review of its use in the treatment of hyperuricaemia in patients with gout. Drugs 75:427–438. CrossRefGoogle Scholar
  12. Ishikawa T, Aw W, Kaneko K (2013) Metabolic interactions of Purine derivatives with human ABC transporter ABCG2: genetic testing to assess gout risk. Pharmaceuticals (Basel) 6:1347–1360. CrossRefGoogle Scholar
  13. Kong LD, Cai Y, Huang WW, Cheng CHK, Tan RX (2000) Inhibition of xanthine oxidase by some Chinese medicinal plants used to treat gout. J Ethnopharmacol 73:199–207. CrossRefGoogle Scholar
  14. La Torre GL, Saitta M, Vilasi F, Pellicanò T, Dugo G (2006) Direct determination of phenolic compounds in Sicilian wines by liquid chromatography with PDA and MS detection. Food Chem 94:640–650. CrossRefGoogle Scholar
  15. Li Q, Ren FQ, Yang CL, Zhou LM, Liu YY, Xiao J, Zhu L, Wang ZG (2015) Anti-proliferation effects of isorhamnetin on lung cancer cells in vitro and in vivo. Asian Pac J Cancer Prev 16:3035–3042. CrossRefGoogle Scholar
  16. Manach C, Morand C, Crespy V, Demigné C, Texier O, Régérat F, Rémésy C (1998) Quercetin is recovered in human plasma as conjugated derivatives which retain antioxidant properties. FEBS Lett 426:331–336. CrossRefGoogle Scholar
  17. Middleton E, Theoharides T, Kandaswami C (2000) The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev 52:673–751Google Scholar
  18. Minakawa M, Kawano A, Miura Y, Yagasaki K (2011) Hypoglycemic effect of resveratrol in type 2 diabetic model db/db mice and its actions in cultured L6 myotubes and RIN-5F pancreatic β-cells. J Clin Biochem Nutr 48:237–244. CrossRefGoogle Scholar
  19. Mo SF, Zhou F, Lv YZ, Hu QH, Zhang DM, Kong LD (2007) Hypouricemic action of selected flavonoids in mice: structure–activity relationships. Biol Pharm Bull 30:1551–1556. CrossRefGoogle Scholar
  20. Mondal A, Rajalingam D, Kumar Maity T (2013) Anti-inflammatory effect of O-methylated flavonol 2-(3,4-dihydroxy-phenyl)- 3,5-dihydroxy-7-methoxy-chromen-4-one obtained from Cassia sophera Linn in rats. J Ethnopharmacol 147:525–9. Google Scholar
  21. Morand C, Crespy V, Manach C, Besson C, Demigné C, Rémésy C (1998) Plasma metabolites of quercetin and their antioxidant properties. Am J Physiol 275:R212–R219. Google Scholar
  22. Morand C, Manach C, Crespy V, Remesy C (2000) Respective bioavailability of quercetin aglycone and its glycosides in a rat model. BioFactors 12:169–174. CrossRefGoogle Scholar
  23. Nessa F, Ismail Z, Mohamed N (2010) Xanthine oxidase inhibitory activities of extracts and flavonoids of the leaves of Blumea balsamifera. Pharm Biol 48:1405–1412. CrossRefGoogle Scholar
  24. Olsson ME, Gustavsson K-E, Vågen IM (2010) Quercetin and isorhamnetin in sweet and red cultivars of onion (Allium cepa L.) at harvest, after field curing, heat treatment, and storage. J Agric Food Chem 58:2323–2330. CrossRefGoogle Scholar
  25. Petrie JL, Patman GL, Sinha I, Alexander TD, Reeves HL, Agius L (2013) The rate of production of uric acid by hepatocytes is a sensitive index of compromised cell ATP homeostasis. Am J Physiol Endocrinol Metab 305:E1255–E1265. CrossRefGoogle Scholar
  26. Phan TT, Wang L, See P, Grayer RJ, Chan SY, Lee ST (2001) Phenolic compounds of Chromolaena odorata protect cultured skin cells from oxidative damage: implication for cutaneous wound healing. Biol Pharm Bull 24:1373–1379. CrossRefGoogle Scholar
  27. Schilaty ND, Hedges DM, Jang EY, Folsom RJ, Yorgason JT, McIntosh JM, Steffensen SC (2014) Acute ethanol inhibits dopamine release in the nucleus accumbens via α6 nicotinic acetylcholine receptors. J Pharmacol Exp Ther 349:559–567. CrossRefGoogle Scholar
  28. Shi Y, Williamson G (2016) Quercetin lowers plasma uric acid in pre-hyperuricaemic males: a randomised, double-blinded, placebo-controlled, cross-over trial. Br J Nutr 115:800–806. CrossRefGoogle Scholar
  29. Teets AS, Minardi CS, Sundararaman M, Hughey CA, Were LM (2009) Extraction, identification, and quantification of flavonoids and phenolic acids in electron beam-irradiated almond skin powder. J Food Sci 74:298–305. CrossRefGoogle Scholar
  30. Thottam GE, Krasnokutsky S, Pillinger MH (2017) Gout and metabolic syndrome: a tangled web. Curr Rheumatol Rep 19:60. CrossRefGoogle Scholar
  31. Tsushima Y, Nishizawa H, Tochino Y, Nakatsuji H, Sekimoto R, Nagao H, Shirakura T, Kato K, Imaizumi K, Takahashi H, Tamura M, Maeda N, Funahashi T, Shimomura I (2013) Uric acid secretion from adipose tissue and its increase in obesity. J Biol Chem 288:27138–27149. CrossRefGoogle Scholar
  32. Wang CP, Wang X, Zhang X, Shi YW, Liu L, Kong LD (2010) Morin improves urate excretion and kidney function through regulation of renal organic ion transporters in hyperuricemic mice. J Pharm Pharm Sci 13:411. CrossRefGoogle Scholar
  33. Zhang BN, Hou YL, Liu BJ, Liu QM, Qiao GF (2010) The Rhododendron dauricum L. flavonoids exert vasodilation and myocardial preservation. Iran J Pharm Res IJPR 9:303–311Google Scholar
  34. Zhang C, Wang R, Zhang G, Gong D (2018) Mechanistic insights into the inhibition of quercetin on xanthine oxidase. Int J Biol Macromol 112:405–412. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Shin-ichi Adachi
    • 1
    Email author
  • Shinji Kondo
    • 1
  • Yusuke Sato
    • 2
  • Fumiaki Yoshizawa
    • 2
    • 3
  • Kazumi Yagasaki
    • 1
  1. 1.Center for Bioscience Research and EducationUtsunomiya UniversityUtsunomiyaJapan
  2. 2.Faculty of AgricultureUtsunomiya UniversityUtsunomiyaJapan
  3. 3.Graduate School of Agricultural ScienceTokyo University of Agriculture and TechnologyTokyoJapan

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