Polyphenol-Enriched Fractions of Cyclopia intermedia Selectively Affect Lipogenesis and Lipolysis in 3T3-L1 Adipocytes

Babalwa U. Jack; Christiaan J. Malherbe; Elize L. Willenburg; Dalene de Beer; Barbara Huisamen; Elizabeth Joubert; Christo J. F. Muller; Johan Louw; Carmen Pheiffer

doi:10.1055/s-0043-119463

Planta Medica, Table of Contents

Planta Med 2018; 84(02): 100-110
DOI: 10.1055/s-0043-119463

Biological and Pharmacological Activity

Original Papers

Georg Thieme Verlag KG Stuttgart · New York

Polyphenol-Enriched Fractions of Cyclopia intermedia Selectively Affect Lipogenesis and Lipolysis in 3T3-L1 Adipocytes

Babalwa U. Jack

¹Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa

²Division of Medical Physiology, Faculty of Medicine and Health Sciences, University of Stellenbosch, Tygerberg, South Africa

,

Christiaan J. Malherbe

³Plant Bioactives Group, Post-Harvest and Agro-Processing Technologies, Agricultural Research Council (ARC), Infruitec-Nietvoorbij, Stellenbosch, South Africa

,

Elize L. Willenburg

³Plant Bioactives Group, Post-Harvest and Agro-Processing Technologies, Agricultural Research Council (ARC), Infruitec-Nietvoorbij, Stellenbosch, South Africa

,

Dalene de Beer

³Plant Bioactives Group, Post-Harvest and Agro-Processing Technologies, Agricultural Research Council (ARC), Infruitec-Nietvoorbij, Stellenbosch, South Africa

⁴Department of Food Science, University of Stellenbosch, Matieland, South Africa

,

Barbara Huisamen

¹Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa

²Division of Medical Physiology, Faculty of Medicine and Health Sciences, University of Stellenbosch, Tygerberg, South Africa

,

Elizabeth Joubert

³Plant Bioactives Group, Post-Harvest and Agro-Processing Technologies, Agricultural Research Council (ARC), Infruitec-Nietvoorbij, Stellenbosch, South Africa

⁴Department of Food Science, University of Stellenbosch, Matieland, South Africa

,

Christo J. F. Muller

¹Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa

²Division of Medical Physiology, Faculty of Medicine and Health Sciences, University of Stellenbosch, Tygerberg, South Africa

⁵Department of Biochemistry and Microbiology, University of Zululand, Kwa-Dlangezwa, South Africa

,

Johan Louw

¹Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa

⁵Department of Biochemistry and Microbiology, University of Zululand, Kwa-Dlangezwa, South Africa

,

Carmen Pheiffer

¹Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa

²Division of Medical Physiology, Faculty of Medicine and Health Sciences, University of Stellenbosch, Tygerberg, South Africa

› Author Affiliations

Abstract

Cyclopia species are increasingly investigated as sources of phenolic compounds with potential as therapeutic agents. Recently, we demonstrated that a crude polyphenol-enriched organic fraction (CPEF) of Cyclopia intermedia, currently forming the bulk of commercial production, decreased lipid content in 3T3-L1 adipocytes and inhibited body weight gain in obese db/db mice. The aim of the present study was to determine whether a more effective product and/or one with higher specificity could be obtained by fractionation of the CPEF by purposely increasing xanthone and benzophenone levels. Fractionation of the CPEF using high performance counter-current chromatography (HPCCC) resulted in four fractions (F1–F4), predominantly containing iriflophenone-3-C-β-D-glucoside-4-O-β-D-glucoside (benzophenone: F1), hesperidin (flavanone: F2), mangiferin (xanthone: F3), and neoponcirin (flavone: F4), as quantified by high-performance liquid chromatography with diode array detection (HPLC-DAD), and confirmed by LC-DAD with mass spectrometric (MS) and tandem MS (MS^E) detection. All fractions inhibited lipid accumulation in 3T3-L1 pre-adipocytes and decreased lipid content in mature 3T3-L1 adipocytes, although their effects were concentration-dependent. F1–F3 stimulated lipolysis in mature adipocytes. Treatment of mature adipocytes with F1 and F2 increased the messenger RNA expression of hormone sensitive lipase, while treatment with F1 and F4 increased uncoupling protein 3 expression. In conclusion, HPCCC resulted in fractions with different phenolic compounds and varying anti-obesity effects. The activities of fractions were lower than the CPEF; thus, fractionation did not enhance activity within a single fraction worthwhile for exploitation as a nutraceutical product, which illustrates the importance of considering synergistic effects in plant extracts.

Key words

Cyclopia intermedia - Fabaceae - HPCCC - phenolic compounds - obesity - type 2 diabetes - lipogenesis - lipolysis

Full Text

References

References
1 Pi-Sunyer X. The medical risks of obesity. Postgrad Med 2009; 121: 21-33
2 Fock KM, Khoo J. Diet and exercise in management of obesity and overweight. J Gastroenterol Hepatol 2013; 28: 59-63
3 Meydani M, Hasan ST. Dietary polyphenols and obesity. Nutrients 2010; 2: 737-751
4 Schutte AL. Systematics of the genus Cyclopia Vent. (Fabaceae, Podalyrieae). Edin B J 1997; 54: 125-170
5 Joubert E, Gelderblom WC, Louw A, de Beer D. South African herbal teas: Aspalathus linearis, Cyclopia spp. and Athrixia phylicoides – a review. J Ethnopharmacol 2008; 119: 376-412
6 Chellan N, Joubert E, Strijdom H, Roux C, Louw J, Muller CJF. Aqueous extract of unfermented honeybush (Cyclopia maculata) attenuates STZ-induced diabetes and beta-cell cytotoxicity. Planta Med 2014; 80: 622-629
7 Muller CJF, Joubert E, Gabuza K, de Beer D, Fey SJ, Louw J. Assessment of the antidiabetic potential of an aqueous extract of honeybush (Cyclopia intermedia) in streptozotocin and obese insulin resistant Wistar rats. In: Rasooli I. ed. Phytochemicals. Bioactives and Impact on Health. Croatia: Intech; 2011: 311-332
8 Joubert E, Richards ES, Merwe JD, de Beer D, Manley M, Gelderblom WCA. Effect of species variation and processing on phenolic composition and in vitro antioxidant activity of aqueous extracts of Cyclopia spp. (honeybush tea). J Agric Food Chem 2008; 56: 954-963
9 van der Merwe JD, Joubert E, Richards ES, Manley M, Snijman PW, Marnewick JL, Gelderblom WCA. A comparative study on the antimutagenic properties of aqueous extracts of Aspalathus linearis (rooibos), different Cyclopia spp. (honeybush) and Camellia sinensis teas. Mutat Res 2006; 611: 42-53
10 Marnewick J, Joubert E, Joseph S, Swanevelder S, Swart P, Gelderblom WCA. Inhibition of tumour promotion in mouse skin by extracts of rooibos (Aspalathus linearis) and honeybush (Cyclopia intermedia), unique South African herbal teas. Cancer Lett 2005; 224: 193-202
11 Mortimer M, Visser K, de Beer D, Joubert E, Louw A. Divide and conquer may not be the optimal approach to retain the desirable estrogenic attributes of the Cyclopia nutraceutical extract, SM6Met. PLoS One 2015; 10: e0132950
12 Visagie A, Kasonga A, Deepak V, Moosa S, Marais S, Kruger CM, Coetzee M. Commercial honeybush (Cyclopia spp.) tea extract inhibits osteoclast formation and bone resorption in RAW264.7 murine macrophages – an in vitro study. Int J Environ Res Public Health 2015; 12: 13779-13793
13 Dudhia Z, Louw J, Muller CJF, Joubert E, de Beer D, Kinnear C, Pheiffer C. Cyclopia maculata and Cyclopia subternata (honeybush tea) inhibits adipogenesis in 3T3-L1 pre-adipocytes. Phytomedicine 2013; 20: 401-408
14 Pheiffer C, Dudhia Z, Louw J, Muller CJF, Joubert E. Cyclopia maculata (honeybush tea) stimulates lipolysis in 3T3-L1 adipocytes. Phytomedicine 2013; 20: 1168-1171
15 Jack BU, Malherbe CJ, Huisamen B, Gabuza K, Mazibuko-Mbeje S, Schulze AE, Joubert E, Muller CJF, Louw J, Pheiffer C. A polyphenol-enriched fraction of Cyclopia intermedia decreases lipid content in 3T3-L1 adipocytes and reduces body weight gain of obese db/db mice. S Afr J Bot 2017; 110: 216-229
16 Zhang Y, Qian Q, Ge D, Li Y, Wang X, Chen Q, Gao X, Wang T. Identification of benzophenone C-glucosides from mango tree leaves and their inhibitory effect on triglyceride accumulation in 3T3-L1 adipocytes. J Agric Food Chem 2011; 59: 11526-11533
17 Subash-Babu P, Alshatwi AA. Evaluation of antiobesity effect of mangiferin in adipogenesis-induced human mesenchymal stem cells by assessing adipogenic genes. J Food Biochem 2015; 39: 28-38
18 Jeon HJ, Seo MJ, Choi HS, Lee OH, Lee BY. Gelidium elegans, an edible red seaweed, and hesperidin inhibit lipid accumulation and production of reactive oxygen species and reactive nitrogen species in 3T3-L1 and RAW264.7 cells. Phytother Res 2014; 28: 1701-1709
19 Beelders T, Brand DJ, de Beer D, Malherbe CJ, Mazibuko SE, Muller CJF, Joubert E. Benzophenone C- and O-glucosides from Cyclopia genistoides (honeybush) inhibit mammalian α-glucosidase. J Nat Prod 2014; 77: 2694-2699
20 Islam MN, Ishita IJ, Jung HA, Choi JS. Vicenin 2 isolated from Artemisia capillaris exhibited potent anti-glycation properties. Food Chem Toxicol 2014; 69: 55-62
21 Ku SK, Bae JS. Vicenin-2 and scolymoside inhibit high-glucose-induced vascular inflammation in vitro and in vivo . Can J Physiol Pharmacol 2016; 94: 287-295
22 Wilmsen PK, Spada DS, Salvador M. Antioxidant activity of the flavonoid hesperidin in chemical and biological systems. J Agric Food Chem 2005; 53: 4757-4761
23 Akiyama S, Katsumata S, Suzuki K, Ishimi Y, Wu J, Uehara M. Dietary hesperidin exerts hypoglycemic and hypolipidemic effects in streptozotocin-induced marginal type 1 diabetic rats. J Clin Biochem Nutr 2010; 46: 87-92
24 Chae BS, Shin TY. Hesperidin ameliorates TNF-α-mediated insulin resistance in differentiated 3T3-L1 cells. Nat Prod Sci 2012; 18: 254-260
25 Peng H, Wei Z, Luo H, Yang Y, Wu Z, Gan L, Yang X. Inhibition of fat accumulation by hesperidin in Caenorhabditis elegans . J Agric Food Chem 2016; 64: 5207-5214
26 Wang X, Hasegawa J, Kitamura Y, Wang Z, Matsuda A, Shinoda W, Kimura K. Effects of hesperidin on the progression of hypercholesterolemia and fatty liver induced by high-cholesterol diet in rats. J Pharmacol Sci 2011; 117: 129-138
27 Ohara T, Muroyama K, Yamamoto Y, Murosaki S. Oral intake of a combination of glucosyl hesperidin and caffeine elicits an anti-obesity effect in healthy, moderately obese subjects: a randomized double-blind placebo-controlled trial. Nutr J 2016; 15: 6
28 Pranakhon R, Aromdee C, Pannangpetch P. Effects of iriflophenone 3-C-β-glucoside on fasting blood glucose level and glucose uptake. Pharmacogn Mag 2015; 11: 82-89
29 Shimada T, Nagai E, Harasawa Y, Watanabe M, Negishi K, Akase T, Sai Y, Miyamoto K, Aburada M. Salacia reticulata inhibits differentiation of 3T3-L1 adipocytes. J Ethnopharmacol 2011; 136: 67-74
30 Yoshikawa M, Shimoda H, Nishida N, Takada M, Matsuda H. Salacia reticulata and its polyphenolic constituents with lipase inhibitory and lipolytic activities have mild antiobesity effects in rats. J Nutr 2002; 132: 1819-1824
31 Reynolds AR. Potential relevance of bell-shaped and U-shaped dose-responses for the therapeutic targeting of angiogenesis in cancer. Dose Response 2010; 8: 253-284
32 Boss O, Samec S, Paoloni-Giacobino A, Rossier C, Dulloo A, Seydoux J, Muzzin P, Giacobino JP. Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression. FEBS Lett 1997; 408: 39-42
33 Hilse KE, Kalinovich AV, Rupprecht A, Smorodchenko A, Zeitz U, Staniek K, Erben RG, Pohl EE. The expression of UCP3 directly correlates to UCP1 abundance in brown adipose tissue. Biochim Biophys Acta 2016; 1857: 72-78
34 Shabalina I, Petrovic N, de Jong JA, Kalinovich A, Cannon B, Nedergaard J. UCP1 in Brite/Beige adipose tissue mitochondria is functionally thermogenic. Cell Rep 2013; 5: 1196-1203
35 Schrauwen P, Hesselink M. UCP2 and UCP3 in muscle controlling body metabolism. J Exp Biol 2002; 205: 2275-2285
36 Duncan RE, Ahmadian M, Jaworski K, Sarkadi-Nagy E, Sul HS. Regulation of lipolysis in adipocytes. Annu Rev Nutr 2007; 27: 79-101
37 Holm C. Molecular mechanisms regulating hormone-sensitive lipase and lipolysis. Biochem Soc Trans 2003; 31: 1120-1124
38 Herranz-López M, Fernández-Arroyo S, Pérez-Sanchez A, Barrajón-Catalán E, Beltrán-Debón R, Menéndez JA, Alonso-Villaverde C, Segura-Carretero A, Joven J, Micol V. Synergism of plant-derived polyphenols in adipogenesis: perspectives and implications. Phytomedicine 2012; 19: 253-261
39 Bartoszewski R, Hering A, Marszall M, Hajduk JS, Bartoszewska S, Kapoor N, Kochan K, Ochocka R. Mangiferin has an additive effect on the apoptotic properties of hesperidin in Cyclopia sp. tea extracts. PLoS One 2014; 9: e92128
40 Bruckbauer A, Zemel MB, Thorpe T, Akula MR, Stuckey AC, Osborne D, Martin EB, Kennel S, Wall JS. Synergistic effects of leucine and resveratrol on insulin sensitivity and fat metabolism in adipocytes and mice. Nutr Metab 2012; 9: 77
41 Bruckbauer A, Zemel MB. Synergistic effects of polyphenols and methylxanthines with leucine on AMPK/Sirtuin-mediated metabolism in muscle cells and adipocytes. PLoS One 2014; 9: e89166
42 de Kok TM, van Breda SG, Manson MM. Mechanisms of combined action of different chemopreventive dietary compounds: a review. Eur J Nutr 2008; 47: 51-59
43 Andersen CL, Jensen JL, Orntoft TF. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 2004; 64: 5245-5250
44 Beelders T, de Beer D, Stander MA, Joubert E. Comprehensive phenolic profiling of Cyclopia genistoides (L.) Vent. by LC-DAD-MS and -MS/MS reveals novel xanthone and benzophenone constituents. Molecules 2014; 19: 11760-11790
45 de Beer D, Schulze AE, Joubert E, de Villiers A, Malherbe CJ, Stander MA. Food ingredient extracts of Cyclopia subternata (honeybush): variation in phenolic composition and antioxidant capacity. Molecules 2012; 17: 14602-14624

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