Anti-adipogenic Effects of Sulforaphane-rich Ingredient with Broccoli Sprout and Mustard Seed in 3T3-L1 Preadipocytes

Abstract

Glucoraphanin (GRA) is a precursor of sulforaphane (SFN), which can be synthesized by the enzyme myrosinase. In this study, we developed and validated HPLC analytical methods for the determination of GRA and SFN in mustard seed powder (MSP), broccoli sprout powder (BSP), and the MSP-BSP mixture powder (MBP), and evaluated their anti-adipogenic effects in 3T3-L1 adipocytes. We found that the analysis methods were suitable for the determination of GRA and SFN in MSP, BSP, and MBP. The content of GRA in BSP was 131.11 ± 1.84 µmol/g, and the content of SFN in MBP was 162.29 ± 1.24 µmol/g. In addition, BSP and MBP effectively decreased lipid accumulation content without any cytotoxicity. Both BSP and MBP significantly inhibited the expression of adipogenic proteins and increased the expression of proteins related to lipolysis and lipid metabolism. BSP and MBP inhibited the expression of adipocyte protein 2 (aP2), CCAAT/enhancer-binding protein-α (C/EBP-α), and peroxisome proliferator-activated receptor-γ (PPAR-γ) in 3T3-L1 adipocytes, and inhibited the expression of fatty acid synthase (FAS) through AMP-activated protein kinase (AMPK). Meanwhile, BSP and MBP also increased the expression of the lipolysis-related proteins, uncoupling protein-1 (UCP-1) and carnitine palmitoyltransferase-1 (CPT-1). Moreover, MBP exerted anti-adipogenic to a greater extent than BSP in 3T3-L1 preadipocytes.

Key words

Sinapis alba L - Brassica oleracea var. italic - Brassicaceae - Sulforaphane - Analytical method validation - Anti-adipogenic

• References

• 1 Chait A, den Hartigh LJ. Adipose tissue distribution, inflammation, and its metabolic consequences, including diabetes and cardiovascular disease. Front Cardiovasc Med 2020; 7: 22
• 2 Haczeyni F, Bell-Anderson KS, Farrell GC. Causes and mechanisms of adipocyte enlargement and adipose expansion. Obes Rev 2018; 19: 406-420
• 3 Fabbrini E, Sullivan S, Klein S. Obesity and nonalcoholic fatty liver disease: Biochemical, metabolic, and clinical implications. Hepatology 2010; 51: 679-689
• 4 Chen X, Gui G, Ji W, Xue Q, Wang C, Li H. The relationship between obesity subtypes based on BMI and cardio-cerebrovascular disease. Hypertens Res 2019; 42: 912-919
• 5 Leggio M, Lombardi M, Caldarone E, Severi P, DʼEmidio S, Armeni M, Bravi V, Bendini MG, Mazza A. The relationship between obesity and hypertension: An updated comprehensive overview on vicious twins. Hypertens Res 2017; 40: 947-963
• 6 Zhang X, Zhang Y, Wang P, Zhang SY, Dong Y, Zeng G, Yan Y, Sun L, Wu Q, Liu H, Liu B, Kong W, Wang X, Jiang C. Adipocyte hypoxia-inducible factor 2α suppresses atherosclerosis by promoting adipose ceramide catabolism. Cell Metab 2019; 30: 937-951
• 7 Abraham B, Sellin JH. Drug-induced diarrhea. Curr Gastroenterol Rep 2007; 9: 365-372
• 8 Filippatos TD, Derdemezis C, Gazi IF, Nakou ES, Mikhailidis DP, Elisaf MS. Orlistat-associated adverse effects and drug interactions. Drug Saf 2008; 31: 53-65
• 9 Wang HN, Xiang JZ, Qi Z, Du M. Plant extracts in prevention of obesity. Crit Rev Food Sci Nut 2022; 62: 1-14
• 10 de Freitas Junior LM, de Almeida jr. EB. Medicinal plants for the treatment of obesity: Ethnopharmacological approach and chemical and biological studies. Am J Transl Res 2017; 9: 2050-2064
• 11 Veiga M, Costa EM, Sliva S, Pintado M. Impact of plant extracts upon human health: A review. Crit Rev Food Sci Nutr 2020; 60: 873-886
• 12 Kim JK, Park SU. Current potential health benefits of sulforaphane. EXCLI J 2016; 15: 571-577
• 13 Greaney AJ, Maier NK, Leppla SH, Moayeri M. Sulforaphane inhibits multiple inflammasomes through an Nrf2-independent mechanism. J Leukoc Biol 2016; 99: 189-199
• 14 Sikdar S, Papadopoulou M, Dubois J. What do we know about sulforaphane protection against photoaging?. J Cosmet Dermatol 2016; 15: 72-77
• 15 Clarke JD, Dashwood RH, Ho E. Multi-targeted prevention of cancer by sulforaphane. Cancer Lett 2008; 269: 291-304
• 16 Li Y, Zhang T, Korkaya H, Liu S, Lee HF, Newman B, Yu Y, Clouthier SG, Schwartz SJ, Wicha MS, Sun D. Sulforaphane, a dietary component of broccoli/broccoli sprouts, inhibits breast cancer stem cells. Clin Cancer Res 2010; 16: 2580-2590
• 17 Senanayake GVK, Banigesh A, Wu L, Lee P, Juurlink BHJ. The dietary phase 2 protein inducer sulforaphane can normalize the kidney epigenome and improve blood pressure in hypertensive rats. Am J Hypertens 2012; 25: 229-235
• 18 Xu Y, Fu JF, Chen JH, Zhang ZW, Zou ZQ, Han LY, Hua QH, Zhao JS, Zhang XH, Shan YJ. Sulforaphane ameliorates glucose intolerance in obese mice via the upregulation of the insulin signaling pathway. Food Funct 2018; 9: 4695-4701
• 19 Yao A, Shen Y, Wang A, Chen S, Zhang H, Chen F, Chen Z, Wei H, Zou Z, Shan Y, Zhang X. Sulforaphane induces apoptosis in adipocytes via Akt/p70s6k1/Bad inhibition and ERK activation. Biochem Biophys Res Commun 2015; 465: 696-770
• 20 Azizi SN, Amiri-Besheli B, Sharifi-Mehr S. The isolation and determination of sulforaphane from broccoli tissues by reverse phase-High Performance Liquid Chromatography. J Chin Chem Soc 2011; 58: 906-910
• 21 Manchali S, Chidambara Murthy KN, Patil BS. Crucial facts about health benefits of popular cruciferous vegetables. J Funct Foods 2012; 4: 94-106
• 22 Houghton CA, Fassett RG, Coombes JS. Sulforaphane and other nutrigenomic Nrf2 activators: Can the clinicianʼs expectation be matched by the reality?. Oxid Med Cell Longev 2016; 2016: 7857186
• 23 Ghawi SK, Methven L, Niranjan K. The potential to intensify sulforaphane formation in cooked broccoli (Brassica oleracea var. italica) using mustard seeds (Sinapis alba). Food Chem 2013; 138: 1734-1741
• 24 Hanschen FS, Klopsch R, Oliviero T, Schreiner M, Verkerk R, Dekker M. Optimizing isothiocyanate formation during enzymatic glucosinolate breakdown by adjusting pH value, temperature and dilution in Brassica vegetables and Arabidopsis thaliana. Sci Rep 2017; 7: 1-15
• 25 Yagishita Y, Fahey JW, Dinkova-Kostova AT, Kensler TW. Broccoli or sulforaphane: Is it the source or dose that matters?. Molecules 2019; 24: 3593
• 26 Cieślik E, Leszczyńska T, Filipiak-Florkiewicz A, Sikora E, Pisulewski PM. Effects of some technological processes on glucosinolate contents in cruciferous vegetables. Food Chem 2007; 105: 976-981
• 27 Gu Y, Guo Q, Zhang L, Chen Z, Han Y, Gu Z. Physiological and biochemical metabolism of germinating broccoli seeds and sprouts. J Agric Food Chem 2012; 60: 209-213
• 28 Piekarska A, Kusznierewicz B, Meller M, Dziedziul K, Namieśnik J, Bartoszek A. Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods. Ind Crops Prod 2013; 50: 58-67
• 29 Shen L, Su G, Wang X, Du Q, Wang K. Endogenous and exogenous enzymolysis of vegetable-sourced glucosinolates and influencing factors. Food Chem 2010; 119: 987-994
• 30 Walfish S. Analytical methods: A statistical perspective on the ICH Q2A and Q2B guidelines for validation of analytical methods. BioPharm Int 2006; 19: 1-6
• 31 Dolan JW. Gradient elution, Part V: Baseline drift problems. Can anything be done to correct for baseline drift in gradient separations?. LCGC North America 2013; 31: 538-542
• 32 De Nicola GR, Rollin P, Mazzon E, Iori R. Novel gram-scale production of enantiopure R-sulforaphane from Tuscan black kale seeds. Molecules 2014; 19: 6975-6986
• 33 Choi KM, Lee YS, Sin DM, Lee S, Lee MK, Lee YM, Hong JT, Yun YP, Yoo HS. Sulforaphane inhibits mitotic clonal expansion during adipogenesis through cell cycle arrest. Obesity (Silver Spring) 2012; 20: 1365-1371
• 34 Kim GH, Ju JY, Chung KS, Cheon SY, Gil TY, Cominguez DC, Cha YY, Lee JH, Roh SS, An HJ. Rice hull extract (RHE) suppresses adiposity in high-fat diet-induced obese mice and inhibits differentiation of 3T3-L1 preadipocytes. Nutrients 2019; 11: 1162
• 35 Lee MS, Kim Y. Mulberry fruit extract ameliorates adipogenesis via increasing AMPK activity and downregulating microRNA-21/143 in 3T3-L1 adipocytes. J Med Food 2020; 23: 266-272
• 36 Park HJ, Cho JY, Kim MK, Koh PO, Cho KW, Kim CH, Lee KS, Chung BY, Kim GS, Cho JH. Anti-obesity effect of Schisandra chinensis in 3T3-L1 cells and high fat diet-induced obese rats. Food Chem 2012; 134: 227-234
• 37 Calzadilla P, Sapochnik D, Cosentino S, Diz V, Dicelio L, Calvo JC, Guerra LN. N-acetylcysteine reduces markers of differentiation in 3T3-L1 adipocytes. Int J Mol Sci 2011; 12: 6936-6951
• 38 Moseti D, Regassa A, Kim WK. Molecular regulation of adipogenesis and potential anti-adipogenic bioactive molecules. Int J Mol Sci 2016; 17: 124
• 39 Lee HG, Lu YA, Li X, Hyun JM, Kim HS, Lee JJ, Kim TH, Kim HM, Kang MC, Jeon AY. Anti-Obesity effects of Grateloupia elliptica, a red seaweed, in mice with high-fat diet-induced obesity via suppression of adipogenic factors in white adipose tissue and increased thermogenic factors in brown adipose tissue. Nutrients 2020; 12: 308
• 40 Lefterova MI, Zhang Y, Steger DJ, Schupp M, Schug J, Cristancho A, Feng D, Zhuo D, Stoeckert jr. CJ, Liu XS, Lazar MA. PPARγ and C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale. Genes Dev 2008; 22: 2941-2952
• 41 Farmer SR. Regulation of PPARγ activity during adipogenesis. Int J Obes (Lond) 2005; 29: 13-16
• 42 Jemai R, Drira R, Makni M, Fetoui H, Sakamoto K. Colocynth (Citrullus colocynthis) seed extracts attenuate adipogenesis by down-regulating PPARγ/SREBP-1c and C/EBPα in 3T3-L1 cells. Food Biosci 2020; 33: 100491
• 43 An Y, Zhang Y, Li C, Qian Q, He W, Wang T. Inhibitory effects of flavonoids from Abelmoschus manihot flowers on triglyceride accumulation in 3T3-L1 adipocytes. Fitoterapia 2011; 82: 595-600
• 44 Tontonoz P, Hu E, Graves RA, Budavari AI, Spiegelman BM. mPPAR gamma 2: Tissue-specific regulator of an adipocyte enhancer. Genes Dev 1994; 8: 1224-1234
• 45 Choi YE, Choi SI, Han X, Men X, Jang GW, Kwon HY, Kang SR, Han JS, Lee OH. Radical scavenging-linked anti-adipogenic activity of Aster Scaber ethanolic extract and its bioactive compound. Antioxidants (Basel) 2020; 9: 1290
• 46 Kawano Y, Cohen DE. Mechanisms of hepatic triglyceride accumulation in non-alcoholic fatty liver disease. J Gastroenterol 2013; 48: 434-441
• 47 Kim N, Nam M, Kang MS, Lee JO, Lee YW, Hwang GS, Kim HS. Piperine regulates UCP1 through the AMPK pathway by generating intracellular lactate production in muscle cells. Sci Rep 2017; 7: 1-13
• 48 Hwang JT, Kim SH, Lee MS, Kim SH, Yang HJ, Kim MJ, Kim HS, Ha J, Kim MS, Kwon DY. Anti-obesity effects of ginsenoside Rh2 are associated with the activation of AMPK signaling pathway in 3T3-L1 adipocyte. Biochem Biophys Res Commun 2007; 364: 1002-1008