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Engineering >> 2022, Volume 9, Issue 2 doi: 10.1016/j.eng.2020.05.023

Functional Metabolomics Reveals that Astragalus Polysaccharides Improve Lipids Metabolism through Microbial Metabolite 2-Hydroxybutyric Acid in Obese Mice

a Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
b Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
c Shanghai Key Laboratory of Diabetes Mellitus and Center for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai 200233, China
d University of Hawaii Cancer Center, Honolulu, HI 96813, USA

# These authors contributed equally to this work.

Received: 2019-12-31 Revised: 2020-05-16 Accepted: 2020-05-25 Available online: 2020-10-16

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Abstract

Polysaccharides are widely present in herbs with multiple activities, especially immunity regulation and metabolic benefits for metabolic disorders. However, the underlying mechanisms are not well understood. Functional metabolomics is increasingly used to investigate systemic effects on the host by identifying metabolites with particular functions. This study explores the mechanisms underlying the metabolic benefits of Astragalus polysaccharides (APS) by adopting a functional metabolomics strategy. The effects of APS were determined in eight-week high-fat diet (HFD)-fed obese mice. Then, gas chromatography–time-of-flight mass spectrometry (GC–TOFMS)-based untargeted metabolomics was performed for an analysis of serum and liver tissues, and liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based targeted metabolomics was performed. The potential functions of the metabolites were tested with in vitro and in vivo models of metabolic disorders. Our results first confirmed the metabolic benefits of APS in obese mice. Then, metabolomics analysis revealed that APS supplementation reversed the HFD-induced metabolic changes, and identified 2-hydroxybutyric acid (2-HB) as a potential functional metabolite for APS activity that was significantly decreased by a HFD and reversed by APS. Further study indicated that 2-HB inhibited oleic acid (OA)-induced triglyceride (TG) accumulation. It was also found to stimulate the expression of proteins in lipid degradation in hepatocytes and TG lipolysis in 3T3-L1 cells. Moreover, it was found to reduce serum TG and regulate the proteins involved in lipid degradation in high-fat and high-sucrose (HFHS)-fed mice. In conclusion, our study demonstrates that the metabolic benefits of APS are at least partially due to 2-HB generation, which modulated lipid metabolism both in vitro and in vivo. Our results also highlight that functional metabolomics is practical for investigating the mechanism underlying the systemic benefits of plant polysaccharides.

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References

[ 1 ] Ford ES, William WHG. Prevalence of the metabolic syndrome among US adults. JAMA Cardiol 2002;287(3):356–9. link1

[ 2 ] Jensen MD, Caruso M, Heiling V, Miles JM. Insulin regulation of lipolysis in nondiabetic and IDDM subjects. Diabetes 1989;38(12):1595–601. link1

[ 3 ] Weiss R, Dziura J, Burgert TS, Tamborlane WV, Taksali SE, Yeckel CW, et al. Obesity and the metabolic syndrome in children and adolescents. New Engl J Med 2004;350:2362–74. link1

[ 4 ] Robert HE, Scott MG, Zimmet PZ. The metabolic syndrome. Lancet 2005;365 (9468):1415–28. link1

[ 5 ] Lewis GF, Uffelman KD, Szeto LW, Weller B, Steiner G. Interaction between free fatty acids and insulin in the acute control of very low density lipoprotein production in humans. J Clin Invest 1995;1(95):158–66. link1

[ 6 ] McCarthy EM, Rinella ME. The role of diet and nutrient composition in nonalcoholic fatty liver disease. J Acad Nutr Diet 2012;112(3):401–9. link1

[ 7 ] Cohen JC, Horton JD, Hobbs HH. Human fatty liver disease: old questions and new insights. Science 2011;332(6037):1519–23. link1

[ 8 ] Younossi ZM. Non-alcoholic fatty liver disease—a global public health perspective. J Hepatol 2019;70(3):531–44. link1

[ 9 ] Narayanaswami V, Dwoskin LP. Obesity: current and potential pharmacotherapeutics and targets. Pharmacol Therapeut 2017;170:116–47. link1

[10] Chan K, Zhang H, Lin Z. An overview on adverse drug reactions to traditional Chinese medicines. Br J Clin Pharmacol 2015;80(4):834–43. link1

[11] Lu Y, Jiang Y, Ling L, Zhang Y, Li H, Chen D. Beneficial effects of Houttuynia cordata polysaccharides on ‘‘two-hit” acute lung injury and endotoxic fever in rats associated with anti-complementary activities. Acta Pharm Sin B 2018;8 (2):218–27. link1

[12] Chang C, Lin C, Lu C, Martel J, Ko Y, Ojcius DM, et al. Ganoderma lucidum reduces obesity in mice by modulating the composition of the gut microbiota. Nat Commun 2015;6(1):7489. link1

[13] Wu T, Lin C, Chang C, Lin T, Martel J, Ko Y, et al. Gut commensal Parabacteroides goldsteinii plays a predominant role in the anti-obesity effects of polysaccharides isolated from Hirsutella sinensis. Gut 2019;68(2):248–62. link1

[14] Fu J, Wang Z, Huang L, Zheng S, Wang D, Chen S, et al. Review of the botanical characteristics, phytochemistry, and pharmacology of Astragalus membranaceus (Huangqi). Phytother Res 2014;28(9):1275–83. link1

[15] Liu P, Zhao H, Luo Y. Anti-aging implications of Astragalus membranaceus (Huangqi): a well-known Chinese tonic. Aging Dis 2017;8(6):868–86. link1

[16] Mao X, Yu F, Wang N, Wu Y, Zou F, Wu K, et al. Hypoglycemic effect of polysaccharide enriched extract of Astragalus membranaceus in diet induced insulin resistant C57BL/6J mice and its potential mechanism. Phytomedicine 2009;16(5):416–25. link1

[17] Huang Y, Tsay H, Lu M, Lin C, Yeh C, Liu H, et al. Astragalus membranaceuspolysaccharides ameliorates obesity, hepatic steatosis, neuroinflammation and cognition impairment without affecting amyloid deposition in metabolically stressed APPswe/PS1dE9 mice. Int J Mol Sci 2017;18 (12):2746–63. link1

[18] Liu M, Qin J, Hao Y, Liu M, Luo J, Luo T, et al. Astragalus polysaccharide suppresses skeletal muscle myostatin expression in diabetes: involvement of ROS-ERK and NF-jB pathways. Oxid Med Cell Longev 2013;2013:1–10. link1

[19] He X, He J, Zheng N, Wang S, Li H. Study on the relationship between weight reduction and intestinal bacterial regulation of Astragalus polysaccharide in obese mice. World J Tradit Chin Med 2016;11(11):2379–84. link1

[20] Newgard CB. Metabolomics and metabolic diseases: where do we stand? Cell Metab 2017;25(1):43–56.

[21] Chávez-Talavera O, Tailleux A, Lefebvre P, Staels B. Bile acid control of metabolism and inflammation in obesity, type 2 diabetes, dyslipidemia, and nonalcoholic fatty liver disease. Gastroenterology 2017;7(152):1679–94. link1

[22] Lampropoulou V, Sergushichev A, Bambouskova M, Nair S, Vincent EE, Loginicheva E, et al. Itaconate links inhibition of succinate dehydrogenase with macrophage metabolic remodeling and regulation of inflammation. Cell Metab 2016;24(1):158–66. link1

[23] McNabney S, Henagan T. Short chain fatty acids in the colon and peripheral tissues: a focus on butyrate, colon cancer, obesity and insulin resistance. Nutrients 2017;9(12):1348. link1

[24] Koh GY, Chou G, Liu Z. Purification of a water extract of Chinese sweet tea plant (Rubus suavissimus S. Lee) by alcohol precipitation. J Agric Food Chem 2009;57(11):5000–6. link1

[25] Lin L, Wang P, Du Z, Wang W, Cong Q, Zheng C, et al. Structural elucidation of a pectin from flowers of Lonicera japonica and its antipancreatic cancer activity. Int J Biol Macromol 2016;88:130–7. link1

[26] Ni Y, Su M, Qiu Y, Jia W, Du X. ADAP-GC 3.0: improved peak detection and deconvolution of co-eluting metabolites from GC/TOF-MS data for metabolomics studies. Anal Chem 2016;88(17):8802–11. link1

[27] Gijs Den B, Aycha B, Albert G, Karen Van E, Rick H, Theo H, et al. Short-chain fatty acids protect against high-fat diet-induced obesity via a PPARcdependent switch from lipogenesis to fat oxidation. Diabetes 2015;64:2398–408. link1

[28] Whelan J, Fritsche K. Linoleic acid. Adv Nutr 2013;4(3):311–2. link1

[29] Gooda Sahib Jambocus N, Saari N, Ismail A, Khatib A, Mahomoodally MF, Abdul HA. An investigation into the antiobesity effects of Morinda citrifolia L. leaf extract in high fat diet induced obese rats using a 1 H NMR metabolomics approach. J Diabetes Res 2016;2016:1–14. link1

[30] Drenick EJ, Alvarez LC, Tamasi GC, Brickman AS. Resistance to symptomatic insulin reactions after fasting. J Clin Invest 1972;51(10):2757–62. link1

[31] Werner E, Froehlich R. The potential role for myoinositol in the prevention of gestational diabetes mellitus. Am J Perinatol 2016;13(33):1236–41. link1

[32] Araya J, Rodrigo R, Videla LA, Thielemann L, Orellana M, Pettinelli P, et al. Increase in long-chain polyunsaturated fatty acid n–6/n–3 ratio in relation to hepatic steatosis in patients with non-alcoholic fatty liver disease. Clin Sci 2004;106(6):635–43. link1

[33] Wada Y, Sakiyama S, Sakai H, Sakane F. Myristic acid enhances diacylglycerol kinase d-dependent glucose uptake in myotubes. Lipids 2016;51(8):897–903. link1

[34] Magnusson M, Wang TJ, Clish C, Engström G, Nilsson P, Gerszten RE, et al. Dimethylglycine deficiency and the development of diabetes. Diabetes 2015;64:3010–6. link1

[35] Carvalho VH, Oliveira AHS, de Oliveira LF, Da Silva RP, Di Mascio P, Gualano B, et al. Exercise and b-alanine supplementation on carnosine–acrolein adduct in skeletal muscle. Redox Biol 2018;18:222–8. link1

[36] Saitoh W, Takada S, Hirao J, Shirai M, Iguchi T, Tsuji M, et al. Plasma citrulline is a sensitive safety biomarker for small intestinal injury in rats. Toxicol Lett 2018;295:416–23. link1

[37] Wei P, Rui T, Ruilan W. Clinical application of plasma citrulline in intestinal damage. J Clin Emerg Med 2018;04(19):274–8. link1

[38] Elliott P, Posma JM, Chan Q, Garcia-Perez I, Wijeyesekera A, Bictash M, et al. Urinary metabolic signatures of human adiposity. Sci Transl Med 2015;7 (285):285ra62. link1

[39] Judith LF, Peter AS, Joseph V, Mark DW, Qian G. Role of carnitine in disease. Nutr Metab 2010;7:30. link1

[40] Zhang LS, Davies SS. Microbial metabolism of dietary components to bioactive metabolites: opportunities for new therapeutic interventions. Genome Med 2016;8(1):46. link1

[41] Wikoff WR, Anfora AT, Liu J, Schultz PG, Lesley SA, Peters EC, et al. Metabolomics analysis reveals large effects of gut microflora on mammalian blood metabolites. Proc Natl Acad Sci USA 2009;106(10):3698–703. link1

[42] Traynor J, Mactier R, Geddes CC, Fox JG. How to measure renal function in clinical practice. BMJ 2006;333(7571):733–7. link1

[43] Mensink RP, Zock PL, Kester AD, Katan MB. Effects of dietary fatty acids and carbohydrates on the ratio of serum total to HDL cholesterol and on serum lipids and apolipoproteins: a meta-analysis of 60 controlled trials. Am J Clin Nutr 2003;77(5):1146–55. link1

[44] Mohnen D. Pectin structure and biosynthesis. Curr Opin Plant Biol 2008;11 (3):266–77. link1

[45] Irino Y, Toh R, Nagao M, Mori T, Honjo T, Shinohara M, et al. 2-Aminobutyric acid modulates glutathione homeostasis in the myocardium. Sci Rep 2016;1 (6):36749. link1

[46] Arthur L, Weber I, Stanley LM, Abdul HA. Reasons for the occurrence of the twenty coded protein amino acid. J Mol Evol 1981;17:273–84. link1

[47] Nelson GJ, Schmidt PC, Bartolini G, Kelley DS, Kyle D. The effect of dietary arachidonic acid on platelet function, platelet fatty acid composition, and blood coagulation in humans. Lipids 1997;32(4):421–5. link1

[48] Ferrucci L, Cherubini A, Bandinelli S, Bartali B, Corsi A, Lauretani F, et al. Relationship of plasma polyunsaturated fatty acids to circulating inflammatory markers. J Clin Endocrinol Metab 2006;91(2):439–46. link1

[49] Yan C, Yusha W, Bing W. Research progress on the neuroprotective effect of docosahexaenoic acid. Pract Med Clin 2015;06(18):721–4. link1

[50] Bell M, Wang H, Chen H, McLenithan JC, Gong DW, Yang RZ, et al. Consequences of lipid droplet coat protein downregulation in liver cells: abnormal lipid droplet metabolism and induction of insulin resistance. Diabetes 2008;57(8):2037–45. link1

[51] Hsiao P, Chiou HC, Jiang H, Lee M, Hsieh T, Kuo K. Pioglitazone enhances cytosolic lipolysis, b-oxidation and autophagy to ameliorate hepatic steatosis. Sci Rep 2017;1(7):9030. link1

[52] He W, Xu Y, Ren X, Xiang D, Lei K, Zhang C, et al. Vitamin E ameliorates lipid metabolism in mice with nonalcoholic fatty liver disease via Nrf2/CES1 signaling pathway. Dig Dis Sci 2019;64(11):3182–91. link1

[53] Odegaard JI, Chawla A. Pleiotropic actions of insulin resistance and inflammation in metabolic homeostasis. Science 2013;339(6116):172–7. link1

[54] Yang SZQQ. MicroRNA-124 negatively regulates LPS-induced TNF-a production in mouse macrophages by decreasing protein stability. Chin J Pharmacol 2016;7(37):889–97. link1

[55] Zhu D, Wang Y, Du Q, Liu Z, Liu X. Cichoric acid reverses insulin resistance and suppresses inflammatory responses in the glucosamine-induced HepG2 cells. J Agric Food Chem 2015;63(51):10903–13. link1

[56] Mi Y, Qi G, Gao Y, Li R, Wang Y, Li X, et al. (–)-Epigallocatechin-3-gallate ameliorates insulin resistance and mitochondrial dysfunction in HepG2 cells: involvement of Bmal1. Mol Nutr Food Res 2017;12(61):1700440. link1

[57] Wu T, Lin C, Chang C, Lin T, Martel J, Ko Y, et al. Gut commensal Parabacteroides goldsteinii plays a predominant role in the anti-obesity effects of polysaccharides isolated from Hirsutella sinensis. Gut 2019;2(68):248–62. link1

[58] Porter NT, Martens EC. The critical roles of polysaccharides in gut microbial ecology and physiology. Annu Rev Microbiol 2017;1(71):349–69. link1

[59] Ussher JR, Elmariah S, Gerszten RE, Dyck JRB. The emerging role of metabolomics in the diagnosis and prognosis of cardiovasculat disease. J Am Coll Cardiol 2016;25(68):2850–70. link1

[60] Lampropoulou V, Sergushichev A, Bambouskova M, Nair S, Vincent EE, Loginicheva E, et al. Itaconate links inhibition of succinate dehydrogenase with macrophage metabolic remodeling and regulation of inflammation. Cell Metab 2016;1(24):158–66. link1

[61] Garaycoechea JI, Crossan GP, Langevin F, Daly M, Arends MJ, Patel KJ. Genotoxic consequences of endogenous aldehydes on mouse haematopoietic stem cell function. Nature 2012;7417(489):571–5. link1

[62] Chin RM, Fu X, Pai MY, Vergnes L, Hwang H, Deng G, et al. The metabolite aketoglutarate extends lifespan by inhibiting ATP synthase and TOR. Nature 2014;510(7505):397–401. link1

[63] Min Y, Guowang X. Current and future perspectives of functional metabolomics in disease studies—a review. Anal Chim Acta 2018;1037:41–54. link1

[64] Wu G. Amino acids: metabolism, functions, and nutrition. Amino Acids 2009;37(1):1–17. link1

[65] Chambers ES, Byrne CS, Morrison DJ, Murphy KG, Preston T, Tedford C, et al. Dietary supplementation with inulin-propionate ester or inulin improves insulin sensitivity in adults with overweight and obesity with distinct effects on the gut microbiota, plasma metabolome and systemic inflammatory responses: a randomised cross-over trial. Gut 2019;68(8):1430–8. link1

[66] Chambers ES, Viardot A, Psichas A, Morrison DJ, Murphy KG, Zac-Varghese SEK, et al. Effects of targeted delivery of propionate to the human colon on appetite regulation, body weight maintenance and adiposity in overweight adults. Gut 2015;64(11):1744–54. link1

[67] Newman JC, Verdin E. b-Hydroxybutyrate: a signaling metabolite. Annu Rev Nutr 2017;37(1):51–76. link1

[68] Zhou D, Chen YW, Zhao ZH, Yang RX, Xin FZ, Liu XL, et al. Sodium butyrate reduces high-fat diet-induced non-alcoholic steatohepatitis through upregulation of hepatic GLP-1R expression. Exp Mol Med 2018;50(12):157. link1

[69] Ferrannini E, Natali A, Camastra S, Nannipieri M, Mari A, Adam KP, et al. Early metabolic markers of the development of dysglycemia and type 2 diabetes and their physiological significance. Diabetes 2013;62(5):1730–7. link1

[70] Lin Z, Vicente Gonçalves CM, Dai L, Lu H, Huang J, Ji H, et al. Exploring metabolic syndrome serum profiling based on gas chromatography mass spectrometry and random forest models. Anal Chim Acta 2014;827:22–7. link1

[71] Landaas S. The formation of 2-hydroxybutyric acid in experimental animals. Clin Chim Acta 1975;58(1):23–32. link1

[72] Adams SH. Emerging perspectives on essential amino acid metabolism in obesity and the insulin-resistant state. Adv Nutr 2011;2(6):445–56. link1

[73] Chaillou S, Champomier-Vergès M, Cornet M, Crutz-Le Coq A, Dudez A, Martin V, et al. The complete genome sequence of the meat-borne lactic acid bacterium Lactobacillus sakei 23K. Nat Biotechnol 2005;23(12): 1527–33. link1

[74] Gao C, Zhang W, Ma C, Liu P, Xu P. Kinetic resolution of 2-hydroxybutanoate racemic mixtures by NAD-independent l-lactate dehydrogenase. Bioresour Technol 2011;102(7):4595–9. link1

[75] Heidelberg JF, Seshadri R, Haveman SA, Hemme CL, Paulsen IT, Kolonay JF, et al. The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. Nat Biotechnol 2004;22 (5):554–9. link1

[76] Kapatral V, Anderson I, Ivanova N, Reznik G, Los T, Lykidis A, et al. Genome sequence and analysis of the oral bacterium Fusobacterium nucleatum strain ATCC 25586. J Bacteriol 2002;184(7):2005–18. link1

[77] Monot M, Boursaux-Eude C, Thibonnier M, Vallenet D, Moszer I, Medigue C, et al. Reannotation of the genome sequence of Clostridium difficile strain 630. J Med Microbiol 2011;60(Pt 8):1193–9. link1

[78] Zheng N, Gu Y, Hong Y, Sheng L, Chen L, Zhang F, et al. Vancomycin pretreatment attenuates acetaminophen-induced liver injury through 2-hydroxybutyric acid. J Pharm Anal. In press.

[79] Savage DB, Petersen KF, Shulman GI. Disordered lipid metabolism and the pathogenesis of insulin resistance. Physiol Rev 2007;87(2):507–20. link1

[80] Silva AR, Ruschel C, Helegda C, Wyse AT, Wannmacher CM, Wajner M, et al. Inhibition of in vitro CO2 production and lipid synthesis by 2-hydroxybutyric acid in rat brain. Braz J Med Biol Res 2001;34(5):627–31. link1

[81] Schott MB, Rasineni K, Weller SG, Schulze RJ, Sletten AC, Casey CA, et al. bAdrenergic induction of lipolysis in hepatocytes is inhibited by ethanol exposure. J Biol Chem 2017;292(28):11815–28. link1

[82] Quiroga AD, Li L, Trötzmüller M, Nelson R, Proctor SD, Köfeler H, et al. Deficiency of carboxylesterase 1/esterase-x results in obesity, hepatic steatosis, and hyperlipidemia. Hepatology 2012;56(6):2188–98. link1

[83] Eizirik DL, Colli ML, Ortis F. The role of inflammation in insulitis and b-cell loss in type 1 diabetes. Nat Rev Endocrinol 2009;5(4):219–26. link1

[84] Glass CK, Olefsky JM. Inflammation and lipid signaling in the etiology of insulin resistance. Cell Metab 2012;15(5):635–45. link1

[85] Lee HY, Birkenfeld AL, Jornayvaz FR, Jurczak MJ, Kanda S, Popov V, et al. Apolipoprotein CIII overexpressing mice are predisposed to diet-induced hepatic steatosis and hepatic insulin resistance. Hepatology 2011;54 (5):1650–60. link1

[86] Shoelson SE. Inflammation and insulin resistance. J Clin Invest 2006;116 (7):1793–801. link1

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