The aim of the present study was to investigate effects of the ethyl acetate fraction of an ethanol extract of Coix lachryma-jobi (ECLJ) on glucose uptake and adipocyte differentiation in 3T3-L1 cells. ECLJ phosphorylated AMP-activated protein kinase (AMPK) and its downstream substrate acetyl-coenzymeA carboxylase in 3T3-L1 cells in a time- and dose-dependent manner. Moreover, we discovered that compound C inhibits ECLJ-stimulated ACC phosphorylation. In addition, ECLJ exhibited a dose-dependent stimulation of glucose uptake in 3T3-L1 cells, and this increase was obviously attenuated by compound C. ECLJ also caused a decrease in the expression levels of adipogenesis factors such as fatty acid synthase, sterol-regulatory-element-binding protein-1c, peroxisome proliferator-activated receptor γ, and CAATT/enhancer binding protein α in a dose-dependent manner. Differentiation was examined by Oil red O staining activity after ECLJ treatment for 6 days. ECLJ decreased mean droplet size. These results suggest a possible role for AMPK in the process of adipose differentiation and that ECLJ targeted for adipocyte functions could be effective in improving the symptoms of metabolic syndrome.
Get full access to this article
View all access options for this article.
References
1.
MollerDE. New drug targets for type-2 diabetes and the metabolism syndrome. Nature, 2001; 414:821–827.
2.
AshrafiK. Obesity and the regulation of fat metabolism. Wormbook (The C. elegans Research Community, ed.)10.1895/wormbook.1.130.1www.wormbook.org. 2007 March 9.
3.
HajerGR, van HaeftenTW, VisserenFL. Adipose tissue dysfunction in obesity, diabetes, and vascular diseases. Eur Heart J, 2008; 29:2959–2971.
4.
ZhangBB, ZhouG, LiC. AMPK: an emerging drug target for diabetes and the metabolic syndrome. Cell Metabolism, 2009; 9:407–416.
5.
KempBE, StapletonD, CampellDJet al.AMP-activated protein kinase, super metabolic regulator. Biochem Soc Trans, 2003; 31:162–168.
6.
MisraP, ChakrabartiR. The role of AMP kinase in diabetes. Indian J Med Res, 2007; 125:389–398.
7.
LongYC, ZierathJR. AMP-activated protein kinase signaling in metabolic regulation. J Clin Invest, 2006; 116:1776–1783.
8.
KimEJ, JungSN, SonKHet al.Antidiabetes and antiobesity effect of cryptotanshinone via activation of AMP-activated protein kinase. Mol Pharmacol, 2007; 72:62–72.
9.
LoiDT. Vietnamese Medicinal Plants and Ingredients. Medical Publishing House: Hanoi, 2004.
10.
NagaoT, OtsukaH, KohdaH, SatoT, YamasakiK. Benzoxazinone from Coix lachryma-jobi L var. ma-yuen. Phytochemistry, 1985; 24:2959–2962.
11.
TakahashiM, KonnoC, HikinoH. Isolation and hypoglycemic activity of coixan A, B, C, glycans of Coix lachrymajobi var. ma-yuen seeds. Planta Med, 1986; 52:64–75.
12.
HuangDW, ChungCP, KuoYH, LinYL, ChiangW. Identification of compounds in adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) seed hull extracts that inhibit lipopolysaccharide-induced inflammation in RAW 264.7 macrophages. J Agric Food Chem, 2009; 57:10651–10657.
13.
HungWC, ChangHC. Methanolic extract of adlay seed suppresses COX-2 expression of human lung cancer cells via inhibition of gene transcription. J Agric Food Chem, 2003; 51:7333–7337.
14.
YuF, GaoJ, ZengY, LiuCX. Inhibition of Coix seed extract on fatty acid synthase, a novel target for anticancer activity. J Ethnopharmacol, 2008; 119:252–258.
15.
WooJH, LiD, WilsbachKet al.Coix seed extract, a commonly used treatment for cancer in China, inhibits NFkappaB and protein kinase C signaling. Cancer Biol Ther, 2007; 6:2005–2011.
16.
HsiaSM, ChiangW, KuoYH, WangPS. Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts. Int J Impotence Res, 2006; 18:264–274.
17.
HsiaSM, YehCL, KuoYH, WP>S, ChiangW. Effects of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) hull extract on the secretion of progesterone and estradiol in vivo and in vitro. Exp Biol Med, 2007; 232:1181–1194.
18.
ChangLL, WunAW, HungCT, HsiaSM, ChiangW, WangPS. Effects of crude adlay hull acetone extract on corticosterone release from rat zona fasciculata-reticularis cells. Naunyn Schmiedebergs Arch Pharmacol, 2006; 374:141–152.
19.
KimSO, YunSJ, JungBet al.Hypolipidemic effects of crude extract of adlay seed (Coix lachrymajobi var. mayuen) in obesity rat fed high fat diet: relations of TNF-alpha and leptin mRNA expressions and serum lipid levels. Life Sci, 2004; 75:1391–1404.
20.
StudentAK, HsuRY, LaneMD. Induction of fatty acid synthase synthesis in differentiating 3 T3 L1 preadipocytes. J Biol Chem, 1980; 255:4745–4750.
21.
KameiR, KitagawaY, KadokuraMet al.Shikonin stimulated glucose uptake in 3T3-L1 adipocytes via an insulin independent tyrosine kinase pathway. Biochem Biophys Res Commun, 2002; 292:642–651.
22.
WuX, WakefieldJK, LiuHet al.Development of a novel trans-lentiviral vector that affords predictable safety. Mol Ther, 2000; 2:47–55.
23.
MosmannT. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 1983; 65:55–63.
24.
HardieDG, CarlingD. The AMP-activated protein kinase-fuel gauge of the mammalian cell?Eur J Biochem, 1997; 246:259–273.
25.
ZouMH, HouXY, ShiCM, NagataD, WalshK, CohenRA. Modulation by peroxynitrite of Akt- and AMP-activated kinase-dependent Ser1179 phosphorylation of endothelial nitric oxide synthase. J Biol Chem, 2002; 277:32552–32557.
26.
KimEK, MillerI, AjaSet al.C75, a fatty acid synthase inhibitor, reduces food intake via hypothalamic AMP-activated protein kinase. J Biol Chem, 2004; 279:19970–19976.
27.
NamM, LeeWH, BaeEJ, KimSG. Compound C inhibits clonal expansion of preadipocytes by increasing p21 level irrespectively of AMPK inhibition. Arch Biochem Biophys, 2008; 249:74–81.
28.
LeeES, UhmKO, LeeYMet al.CAPE (caffeic acid phenethyl ester) stimulates glucose uptake through AMPK (AMP-activated protein kinase) activation in skeletal muscle cells. Biochem Biophys Res Commun, 2007; 361:854–858.
29.
HardieDG. Minireview: the AMP-activated protein kinase cascade: the key sensor of cellular energy status. Endocrinology, 2003; 144:5179–5183.
30.
CarlingD. The AMP-activated protein kinase cascade—a unifying system for energy control. Trends Biochem Sci, 2004; 29:18–24.
31.
HardieDG, HawleySA. AMP-activated protein kinase: the energy charge hypothesis revisited. Bioessays, 2001; 23:1112–1119.
32.
MerrillGF, KurthEJ, HardieDG, WinderWW. AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle. Am J Physiol, 1997; 273:E1107–E1112.
33.
Kurth-KraczekEJ, HirshmanMF, GoodyearLJ, WinderWW. 5-AMP activated protein kinase activation causes GLUT4 translocation in skeletal muscle. Diabetes, 1999; 48:1667–1671.
34.
CarlingD, ZammitVA, HardieDG. A common bicyclic protein kinase cascade inactivates the regulatory enzyme of fatty acid and cholesterol biosynthesis. FEBS Lett, 1987; 223:217–222.
35.
SalahuddinM, JalalpureSS. Antidiabetic activity of aqueous fruit extract of Cucumis trigonus Roxb. in streptozotocin-induced-diabetic rats. J Ethnopharmacol, 2009; 127:565–567.
36.
Alonso-CastroAJ, Zapata-BustosR, Romo-YañezJ, Camarillo-LedesmaP, Gómez-SánchezM, Salazar-OlivoLA. The antidiabetic plants Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae) and Teucrium cubense Jacq (Lamiaceae) induce the incorporation of glucose in insulin-sensitive and insulin-resistant murine and human adipocytes. J Ethnopharmacol, 2009; 127:1–6.
37.
RayalamS, YangJY, Della-FeraMA, ParkHJ, AmbatiS, BaileCA. Anti-obesity effects of xanthohumol plus guggulsterone in 3T3-L1 adipocytes. J Med Food, 2009; 12:846–853.
38.
HongJH, HwangEY, KimHJ, JeongYJ, LeeIS. Artemisia capillaris inhibits lipid accumulation in 3T3-L1 adipocytes and obesity in C57BL/6J mice fed a high fat diet. J Med Food, 2009; 12:736–745.
39.
HayashiT, HirshmanMF, FujiiN, HabinowskiSA, WittersLA, GoodyearLJ. Metabolic stress and altered glucose transport: activation of AMP-activated protein kinase as a unifying coupling mechanism. Diabetes, 2000; 49:527–531.
40.
HanSM, NamkoongC, JangPGet al.Hypothalamic AMP-activated protein kinase mediates counter-regulatory responses to hypoglycaemia in rats. Diabetologia, 2005; 48:2170–2178.
KefasBA, CaiY, KerckhofsKet al.Metformin-induced stimulation of AMP activated protein kinase in beta-cells impairs their glucose responsiveness and can lead to apoptosis. Biochem Pharmacol, 2004; 68:409–416.
43.
CollinsQF, LiuHY, PiJ, LiuZ, QuonMJ, CaoW. Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, suppresses hepatic gluconeogenesis through 5′-AMP-activated protein kinase. J Biol Chem, 2007; 282:30143–30149.
44.
HormanS, BrowneG, KrauseUet al.Activation of AMP-activated protein kinase leads to the phosphorylation of elongation factor 2 and an inhibition of protein synthesis. Curr Biol, 2002; 12:1419–1423.
45.
ClaycombeKJ, JonesBH, StandridgeMKet al.Insulin increases fatty acid synthase gene transcription in human adipocytes. Am J Physiol, 1998; 274:R1253–R1259.
46.
PaulauskisJD, SulHS. Hormonal regulation of mouse fatty acid synthase gene transcription in liver. J Biol Chem, 1989; 264:574–577.
WoodsA, Azzout-MarnicheD, FeretzMet al.Characterization of the role of AMP-activated protein kinase in the regulation of glucose-activated gene expression using constitutively active and dominant negative forms of the kinase. Mol Cell Biol, 2000; 20:6704–6711.
49.
KimJB, SpiegelmanBM. ADD1/SREBP1 promotes adipocyte differentiation and gene expression linked to fatty acid metabolism. Genes Dev, 1996; 10:1096–1107.
AhnJ, LeeH, KimS, ParkJ, HaT. The anti-obesity effect of quercetin is mediated by the AMPK and MAPK signaling pathways. Biochem Biophys Res Commun, 2008; 373:545–549.
52.
MoonHS, ChungCS, LeeHG, KimTG, ChoiYJ, ChoCS. Inhibitory effect of (−)-epigallocatechin-3-gallate on lipid accumulation of 3T3-L1 cells. Obesity, 2007; 15:2571–2582.
53.
KimSH, ParkHS, LeeMSet al.Vitisin A inhibits adipocyte differentiation through cell cycle arrest in 3T3-L1 cells. Biochem Biophys Res Commun, 2008; 372:108–113.
54.
HuangDW, KuoYH, LinFY, LinYL, ChiangW. Effect of Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) Testa and its phenolic components on Cu2+-treated low-density lipoprotein (LDL) oxidation and lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophages. J Agric Food Chem, 2009; 57:2259–2266.
55.
HsiaSM, TsengYW, WangSWet al.Effect of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) hull extracts on testosterone release from rat Leydig cells. Phytother Res, 2009; 23:687–695.
56.
WuTT, CharlesAL, HuangTC. Determination of the contents of the main biochemical compounds of Adlay (Coxi lachrymal-jobi)Food Chem, 2007; 104:1509–1515.
57.
HsuCL, YenGC. Effects of flavonoids and phenolic acids on the inhibition of adipogenesis in 3T3-L1 adipocytes. J Agric Food Chem, 2007; 55:8404–8410.
58.
PrabhakarPK, DobleM. Synergistic effect of phytochemicals in combination with hypoglycemic drugs on glucose uptake in myotubes. Phytomedicine, 2009; 16:1119–1126.
59.
HsuCL, YenGC. Phenolic compounds: evidence for inhibitory effects against obesity and their underlying molecular signaling mechanisms. Mol Nutr Food Res, 2008; 52:53–61.
60.
WelschCA, LachancePA, WassermanBP. Dietary phenolic compounds: inhibition of Na+-dependent d-glucose uptake in rat intestinal brush border membrane vesicles. J Nutr, 1989; 119:1698–1704.
61.
JohnstonK, SharpP, CliffordM, MorganL. Dietary polyphenols decrease glucose uptake by human intestinal Caco-2 cells. FEBS Lett, 2005; 579:1653–1657.
