The purpose of this study is to prove the lipid-regulating effects of neohesperidin (NHP) and explore the potential mechanisms related to fibroblast growth factor 21 (FGF21) and AMP-activated protein kinase (AMPK). Free fatty acids (FFAs)-induced lipid-accumulated HepG2 cells, acutely egg yolk-induced dyslipidemia and chronically diet-induced obese (DIO) model mice were treated with NHP. Biochemical analyses were carried out to determine the lipid profiles. Western blotting and real-time PCR were employed to analyze FGF21, AMPK and the related proteins or mRNA expressions. Body weight and food intake were measured in DIO mice. siRNA or inhibitors of FGF21 or AMPK were utilized in further study. NHP showed potent hypolipidemic effect in HepG2 cells loaded with FFAs and reversed the pathological changes of lipid in the acute or chronic dyslipidemia mouse model. It obviously improved the lipid profiles in plasma, liver and gastrocnemius muscles in DIO mice, and led to a significant body weight loss. Simultaneously, FGF21 protein expression or secretion, and AMPK/sirtuin type 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) axis or related molecules, was improved by NHP in HepG2 cells and/or DIO mice. Furthermore, the siRNA or inhibitor targeting FGF21 or AMPK rejected the triglyceride-lowering effect of NHP. In conclusion, NHP regulates lipid metabolism in vivo and in vitro via FGF21 and AMPK/SIRT1/PGC-1α signaling axis.

Wu L, Parhofer KG: Diabetic dyslipidemia. Metabolism 2014;63:1469-1479.
Bell DS, Al Badarin F, O'Keefe JH Jr: Therapies for diabetic dyslipidaemia. Diabetes Obes Metab 2011;13:313-325.
Safrano L, Bourneau-Martin D, Le Clech C, Chennebault JM, Jamet A, Drablier G, Lagarce L, Lainé-Cessac P: Suspicion of fenofibrate-related drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome: a case report. Therapie 2014;69:461-464.
Stroes ES, Thompson PD, Corsini A, Vladutiu GD, Raal FJ, Ray KK, Roden M, Stein E, Tokgözoğlu L, Nordestgaard BG, Bruckert E, De Backer G, Krauss RM, Laufs U, Santos RD, Hegele RA, Hovingh GK, Leiter LA, Mach F, März W, Newman CB, Wiklund O, Jacobson TA, Catapano AL, Chapman MJ, Ginsberg HN: Statin-associated muscle symptoms: impact on statin therapy-European atherosclerosis society consensus panel statement on assessment, aetiology and management. Eur Heart J 2015;36:1012-1022.
Wright RS: Recent clinical trials evaluating benefit of drug therapy for modification of HDL cholesterol. Curr Opin Cardiol 2013;28:389-398.
Wong CP, Kaneda T, Morita H: Plant natural products as an anti-lipid droplets accumulation agent. J Nat Med 2014;68:253-266.
Toth A, Toth G, Kery A: Polyphenol composition and antioxidant capacity of three Lysimachia species. Nat Prod Commun 2014;9:1473-1478.
Kozłowska A, Szostak-Wegierek D: Flavonoids - food sources and health benefits. Rocz Panstw Zakl Hig 2014;65:79-85.
Alam MA, Subhan N, Rahman MM, Uddin SJ, Reza HM, Sarker SD: Effect of citrus flavonoids, naringin and naringenin, on metabolic syndrome and their mechanisms of action. Adv Nutr 2014;5:404-417.
Nogata Y, Sakamoto K, Shiratsuchi H, Ishii T, Yano M, Ohta H: Flavonoid composition of fruit tissues of citrus species. Biosci Biotechnol Biochem 2006;70:178-192.
Jung UJ, Kim HJ, Lee JS, Lee MK, Kim HO, Park EJ, Kim HK, Jeong TS, Choi MS: Naringin supplementation lowers plasma lipids and enhances erythrocyte antioxidant enzyme activities in hypercholesterolemic subjects. Clin Nutr 2003;22:561-568.
Jung UJ, Lee MK, Jeong KS, Choi MS: The hypoglycemic effects of hesperidin and naringin are partly mediated by hepatic glucose-regulating enzymes in C57BL/KsJ-db/db mice. J Nutr 2004;134:2499-2503.
Zhang J, Sun C, Yan Y, Chen Q, Luo F, Zhu X, Li X, Chen K: Purification of naringin and neohesperidin from Huyou (Citrus changshanensis) fruit and their effects on glucose consumption in human HepG2 cells. Food Chem 2012;135:1471-1478.
Jia S, Hu Y, Zhang W, Zhao X, Chen Y, Sun C, Li X, Chen K: Hypoglycemic and hypolipidemic effects of neohesperidin derived from citrus aurantium L. In diabetic KK-A(y) mice. Food Funct 2015;6:878-886.
Adams AC, Astapova I, Fisher FM, Badman MK, Kurgansky KE, Flier JS, Hollenberg AN, Maratos-Flier E: Thyroid hormone regulates hepatic expression of fibroblast growth factor 21 in a PPARalpha-dependent manner. J Biol Chem 2010;285:14078-14082.
Dostálová I, Haluzíková D, Haluzík M: Fibroblast growth factor 21: a novel metabolic regulator with potential therapeutic properties in obesity/type 2 diabetes mellitus. Physiol Res 2009;58:1-7.
Li H, Zhang J, Jia W: Fibroblast growth factor 21: a novel metabolic regulator from pharmacology to physiology. Front Med 2013;7:25-30.
Cuevas-Ramos D, Aguilar-Salinas CA, Gómez-Pérez FJ: Metabolic actions of fibroblast growth factor 21. Curr Opin Pediatr 2012;24:523-529.
Zhang BB, Zhou G, Li C: AMPK: an emerging drug target for diabetes and the metabolic syndrome. Cell Metab 2009;9:407-416.
Dahlhoff C, Worsch S, Sailer M, Hummel BA, Fiamoncini J, Uebel K, Obeid R, Scherling C, Geisel J, Bader BL, Daniel H: Methyl-donor supplementation in obese mice prevents the progression of NAFLD, activates AMPK and decreases acyl-carnitine levels. Mol Metab 2014;3:565-580.
Foretz M, Ancellin N, Andreelli F, Saintillan Y, Grondin P, Kahn A, Thorens B, Vaulont S, Viollet B: Short-term overexpression of a constitutively active form of AMP-activated protein kinase in the liver leads to mild hypoglycemia and fatty liver. Diabetes 2005;54:1331-1339.
Chau MD, Gao J, Yang Q, Wu Z, Gromada J: Fibroblast growth factor 21 regulates energy metabolism by activating the AMPK-SIRT1-PGC-1alpha pathway. Proc Natl Acad Sci U S A 2010;107:12553-12558.
Si M, Yan Y, Tang L, Wu H, Yang B, He Q, Wu H: A novel indole derivative compound GY3 improves glucose and lipid metabolism via activation of AMP-activated protein kinase pathway. Eur J Pharmacol 2013;698:480-488.
Cousin SP, Hügl SR, Wrede CE, Kajio H, Myers MG Jr, Rhodes CJ: Free fatty acid-induced inhibition of glucose and insulin-like growth factor I-induced deoxyribonucleic acid synthesis in the pancreatic beta-cell line INS-1. Endocrinology 2001;142:229-240.
McGrath JC, Drummond GB, McLachlan EM, Kilkenny C, Wainwright CL: Guidelines for reporting experiments involving animals: the ARRIVE guidelines. Br J Pharmacol 2010;160:1573-1576.
Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG: Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 2010;160:1577-1579.
Lin S, Thomas TC, Storlien LH, Huang XF: Development of high fat diet-induced obesity and leptin resistance in C57Bl/6J mice. Int J Obes Relat Metab Disord 2000;24:639-646.
Assini JM, Mulvihill EE, Huff MW: Citrus flavonoids and lipid metabolism. Curr Opin Lipidol 2013;24:34-40.
Jung UJ, Lee MK, Park YB, Kang MA, Choi MS: Effect of citrus flavonoids on lipid metabolism and glucose-regulating enzyme mRNA levels in type-2 diabetic mice. Int J Biochem Cell Biol 2006;38:1134-1145.
Mulvihill EE, Allister EM, Sutherland BG, Telford DE, Sawyez CG, Edwards JY, Markle JM, Hegele RA, Huff MW: Naringenin prevents dyslipidemia, apolipoprotein B overproduction, and hyperinsulinemia in LDL receptor-null mice with diet-induced insulin resistance. Diabetes 2009;58:2198-2210.
Mulvihill EE, Assini JM, Lee JK, Allister EM, Sutherland BG, Koppes JB, Sawyez CG, Edwards JY, Telford DE, Charbonneau A, St-Pierre P, Marette A, Huff MW: Nobiletin attenuates VLDL overproduction, dyslipidemia, and atherosclerosis in mice with diet-induced insulin resistance. Diabetes 2011;60:1446-1457.
Liem AH, van de Woestijne AP, Roeters van Lennep HW, Zwinderman AH, van der Steeg WA, Jukema JW: ApoB/A1 and LDL-C/HDL-C and the prediction of cardiovascular risk in statin-treated patients. Curr Med Res Opin 2008;24:359-364.
Sellers EA, Singh GR, Sayers SM: Apo-B/AI ratio identifies cardiovascular risk in childhood: the Australian aboriginal birth cohort study. Early Hum Dev 2007;83:S128-S129.
Inagaki T, Dutchak P, Zhao G, Ding X, Gautron L, Parameswara V, Li Y, Goetz R, Mohammadi M, Esser V, Elmquist JK, Gerard RD, Burgess SC, Hammer RE, Mangelsdorf DJ, Kliewer SA: Endocrine regulation of the fasting response by PPARalpha-mediated induction of fibroblast growth factor 21. Cell Metab 2007;5:415-425.
Badman MK, Pissios P, Kennedy AR, Koukos G, Flier JS, Maratos-Flier E: Hepatic fibroblast growth factor 21 is regulated by PPARalpha and is a key mediator of hepatic lipid metabolism in ketotic states. Cell Metab 2007;5:426-437.
Cicione C, Degirolamo C, Moschetta A: Emerging role of fibroblast growth factors 15/19 and 21 as metabolic integrators in the liver. Hepatology 2012;56:2404-2411.
Hindricks J, Ebert T, Bachmann A, Kralisch S, Lössner U, Kratzsch J, Stolzenburg JU, Dietel A, Beige J, Anders M, Bast I, Blüher M, Stumvoll M, Fasshauer M: Serum levels of fibroblast growth factor-21 are increased in chronic and acute renal dysfunction. Clin Endocrinol (Oxf) 2014;80:918-924.
Feng WK, Liu YL, Zhao CQ, Falkner KC, Mohammad MK, Cave MC, McClain CJ: Serum and hepatic fibroblast growth factor 21 (FGF-21) levels are increased in subjects with severe acute alcoholic hepatitis and in mice exposed to chronic-binge alcohol by decreased transcriptional suppression. Hepatology 2013;58:840a.
Li H, Fang Q, Gao F, Fan J, Zhou J, Wang X, Zhang H, Pan X, Bao Y, Xiang K, Xu A, Jia W: Fibroblast growth factor 21 levels are increased in nonalcoholic fatty liver disease patients and are correlated with hepatic triglyceride. J Hepatol 2010;53:934-940.
Kharitonenkov A, Shiyanova TL, Koester A, Ford AM, Micanovic R, Galbreath EJ, Sandusky GE, Hammond LJ, Moyers JS, Owens RA, Gromada J, Brozinick JT, Hawkins ED, Wroblewski VJ, Li DS, Mehrbod F, Jaskunas SR, Shanafelt AB: FGF-21 as a novel metabolic regulator. J Clin Invest 2005;115:1627-1635.
Zhu S, Ma L, Wu Y, Ye X, Zhang T, Zhang Q, Rasoul LM, Liu Y, Guo M, Zhou B, Ren G, Li D: FGF21 treatment ameliorates alcoholic fatty liver through activation of AMPK-SIRT1 pathway. Acta Biochim Biophys Sin (Shanghai) 2014;46:1041-1048.
Lin X, Li G, He X, Ma X, Zhang K, Zhang H, Zeng G, Wang Z: FGF21 inhibits apolipoprotein(a) expression in HepG2 cells via the FGFR1-ERK1/2-ELK-1 pathway. Mol Cell Biochem 2014;393:33-42.
Fisher FM, Chui PC, Antonellis PJ, Bina HA, Kharitonenkov A, Flier JS, Maratos-Flier E: Obesity is a fibroblast growth factor 21 (FGF21)-resistant state. Diabetes 2010;59:2781-2789.
Fisher FM, Estall JL, Adams AC, Antonellis PJ, Bina HA, Flier JS, Kharitonenkov A, Spiegelman BM, Maratos-Flier E: Integrated regulation of hepatic metabolism by fibroblast growth factor 21 (FGF21) in vivo. Endocrinology 2011;152:2996-3004.
Cantó C, Gerhart-Hines Z, Feige JN, Lagouge M, Noriega L, Milne JC, Elliott PJ, Puigserver P, Auwerx J: AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature 2009;458:1056-1060.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.