Nonalcoholic fatty liver disease is a progressive disease involving the accumulation of lipid droplets in the liver. In this study, we investigated the anti-hepatosteatosis effects of fermented Cordyceps militaris extract (CME) in AML-12 hepatocytes. Although the levels of adenosine and cordycepin were reduced in the extracts of CM grown on germinated soybean (GSCE) and fermented CM grown on germinated soybean (GSC) by Pediococcus pentosaceus ON188 (ON188E), the expression of fatty acid oxidation (FAO) genes were upregulated only by GSC-ON188E treatment in a dose-dependent manner. In contrast, a lipogenic gene, stearoyl Coenzyme A desaturase 1, was downregulated by ON188E. Formation of intracellular lipid droplets by the addition of oleic acid was reduced by ON188E to levels observed in WY14643-treated cells. When cells were treated with ON188E, sphingosine kinase 2 mainly responsible for hepatic sphingosine 1-phosphate (S1P) synthesis was upregulated and S1P was elevated. Collectively, the fermented GSC extract activates FAO through elevation of S1P synthesis and has potential as a therapeutic for hepatosteatosis.
Get full access to this article
View all access options for this article.
References
1.
SanyalAJ, American Gastroenterological Association: AGA technical review on nonalcoholic fatty liver disease. Gastroenterology, 2002; 123:1705–1725.
2.
KooSH, FlechnerL, QiL, et al.: The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism. Nature, 2005; 437:1109–1114.
3.
PosticC, GirardJ: Contribution of de novo fatty acid synthesis to hepatic steatosis and insulin resistance: Lessons from genetically engineered mice. J Clin Invest, 2008; 118:829–838.
4.
LeeSY, HongIK, KimBR, et al.: Activation of sphingosine kinase 2 by endoplasmic reticulum stress ameliorates hepatic steatosis and insulin resistance in mice. Hepatology, 2015; 62:135–146.
5.
SafaeiA, Arefi OskouieA, MohebbiSR, et al.: Metabolomic analysis of human cirrhosis, hepatocellular carcinoma, non-alcoholic fatty liver disease and non-alcoholic steatohepatitis diseases. Gastroenterol Hepatol Bed Bench, 2016; 9:158–173.
6.
Jambor de SousaUL, KossMD, FilliesM, et al.: CPT1alpha over-expression increases long-chain fatty acid oxidation and reduces cell viability with incremental palmitic acid concentration in 293T cells. Biochem Biophys Res Commun, 2005; 338:757–761.
ZengJ, DengS, WangY, LiP, TangL, PangY: Specific inhibition of acyl-CoA oxidase-1 by an acetylenic acid improves hepatic lipid and reactive oxygen species (ROS) metabolism in rats fed a high fat diet. J Biol Chem, 2017; 292:3800–3809.
9.
HashimotoT, CookWS, QiC, YeldandiAV, ReddyJK, RaoMS: Defect in peroxisome proliferator-activated receptor alpha-inducible fatty acid oxidation determines the severity of hepatic steatosis in response to fasting. J Biol Chem, 2000; 275:28918–28928.
10.
HuangJ, JiaY, FuT, et al.: Sustained activation of PPARalpha by endogenous ligands increases hepatic fatty acid oxidation and prevents obesity in ob/ob mice. FASEB J, 2012; 26:628–638.
11.
EnochHG, CatalaA, StrittmatterP: Mechanism of rat liver microsomal stearyl-CoA desaturase. Studies of the substrate specificity, enzyme-substrate interactions, and the function of lipid. J Biol Chem, 1976; 251:5095–5103.
12.
MiyazakiM, KimYC, Gray-KellerMP, AttieAD, NtambiJM: The biosynthesis of hepatic cholesterol esters and triglycerides is impaired in mice with a disruption of the gene for stearoyl-CoA desaturase 1. J Biol Chem, 2000; 275:30132–30138.
13.
DasSK, MasudaM, SakuraiA, SakakibaraM: Medicinal uses of the mushroom Cordyceps militaris: Current state and prospects. Fitoterapia, 2010; 81:961–968.
14.
HanES, OhJY, ParkHJ: Cordyceps militaris extract suppresses dextran sodium sulfate-induced acute colitis in mice and production of inflammatory mediators from macrophages and mast cells. J Ethnopharmacol, 2011; 134:703–710.
15.
ReisFS, BarrosL, CalhelhaRC, et al.: The methanolic extract of Cordyceps militaris (L.) Link fruiting body shows antioxidant, antibacterial, antifungal and antihuman tumor cell lines properties. Food Chem Toxicol, 2013; 62:91–98.
16.
RaoYK, FangSH, WuWS, TzengYM: Constituents isolated from Cordyceps militaris suppress enhanced inflammatory mediator's production and human cancer cell proliferation. J Ethnopharmacol, 2010; 131:363–367.
17.
KimCS, LeeSY, ChoSH, et al.: Cordyceps militaris induces the IL-18 expression via its promoter activation for IFN-gamma production. J Ethnopharmacol, 2008; 120:366–371.
18.
ChangY, JengKC, HuangKF, et al.: Effect of Cordyceps militaris supplementation on sperm production, sperm motility and hormones in Sprague-Dawley rats. Am J Chin Med, 2008; 36:849–859.
19.
LiXT, LiHC, LiCB, DouDQ, GaoMB: Protective effects on mitochondria and anti-aging activity of polysaccharides from cultivated fruiting bodies of Cordyceps militaris. Am J Chin Med, 2010; 38:1093–1106.
20.
HurH: Chemical Ingredients of Cordyceps militaris. Mycobiology, 2008; 36:233–235.
21.
RobsonSC, SchuppanD: Adenosine: Tipping the balance towards hepatic steatosis and fibrosis. J Hepatol, 2010; 52:941–943.
22.
KoupenovaM, Johnston-CoxH, VezeridisA, et al.: A2b adenosine receptor regulates hyperlipidemia and atherosclerosis. Circulation, 2012; 125:354–363.
23.
WuC, GuoY, SuY, et al.: Cordycepin activates AMP-activated protein kinase (AMPK) via interaction with the gamma1 subunit. J Cell Mol Med, 2014; 18:293–304.
24.
KimHG, ShresthaB, LimSY, et al.: Cordycepin inhibits lipopolysaccharide-induced inflammation by the suppression of NF-kappaB through Akt and p38 inhibition in RAW 264.7 macrophage cells. Eur J Pharmacol, 2006; 545:192–199.
25.
LeeSY, DebnathT, KimSK, LimBO: Anti-cancer effect and apoptosis induction of cordycepin through DR3 pathway in the human colonic cancer cell HT-29. Food Chem Toxicol, 2013; 60:439–447.
26.
ShinS, MoonS, ParkY, et al.: Role of cordycepin and adenosine on the phenotypic switch of macrophages via induced anti-inflammatory cytokines. Immune Netw, 2009; 9:255–264.
27.
ChoiHN, JangYH, KimMJ, et al.: Cordyceps militaris alleviates non-alcoholic fatty liver disease in ob/ob mice. Nutr Res Pract, 2014; 8:172–176.
28.
LeeDY, ParkJJ, BarupalDK, FiehnO: System response of metabolic networks in Chlamydomonas reinhardtii to total available ammonium. Mol Cell Proteomics, 2012; 11:973–988.
29.
LeeJE, ChoYU, KimKH, LeeDY: Distinctive metabolomic responses of Chlamydomonas reinhardtii to the chemical elicitation by methyl jasmonate and salicylic acid. Process Biochem, 2016; 51:1147–1154.
30.
LeeJE, LeeYH, KimSY, et al.: Systematic biomarker discovery and coordinative validation for different primary nephrotic syndromes using gas chromatography-mass spectrometry. J Chromatogr A, 2016; 1453:105–115.
31.
YooHH, SonJ, KimDH: Liquid chromatography-tandem mass spectrometric determination of ceramides and related lipid species in cellular extracts. J Chromatogr B Analyt Technol Biomed Life Sci, 2006; 843:327–333.
32.
CuiW, ChenSL, HuKQ: Quantification and mechanisms of oleic acid-induced steatosis in HepG2 cells. Am J Transl Res, 2010; 2:95–104.
33.
Perez-CarrerasM, Del HoyoP, MartinMA, et al.: Defective hepatic mitochondrial respiratory chain in patients with nonalcoholic steatohepatitis. Hepatology, 2003; 38:999–1007.
34.
FujiiH, KawadaN: Inflammation and fibrogenesis in steatohepatitis. J Gastroenterol, 2012; 47:215–225.
35.
PengZ, BoreaPA, VaraniK, et al.: Adenosine signaling contributes to ethanol-induced fatty liver in mice. J Clin Invest, 2009; 119:582–594.
HuangX, CaiW, XuB: Kinetic changes of nutrients and antioxidant capacities of germinated soybean (Glycine max L.) and mung bean (Vigna radiata L.) with germination time. Food Chem, 2014; 143:268–276.
38.
LinPY, LaiHM: Bioactive compounds in legumes and their germinated products. J Agric Food Chem, 2006; 54:3807–3814.
39.
MuhialdinBJ, HassanZ, SadonS: Antifungal activity of Lactobacillus fermentum Te007, Pediococcus pentosaceus Te010, Lactobacillus pentosus G004, and L. paracasi D5 on selected foods. J Food Sci, 2011; 76:M493–M499.
40.
FranciosiE, CarafaI, NardinT, et al.: Biodiversity and gamma-aminobutyric acid production by lactic acid bacteria isolated from traditional alpine raw cow's milk cheeses. Biomed Res Int, 2015; 2015:625740.
41.
NtambiJM, MiyazakiM, StoehrJP, et al.: Loss of stearoyl-CoA desaturase-1 function protects mice against adiposity. Proc Natl Acad Sci U S A, 2002; 99:11482–11486.
42.
MiyazakiM, FlowersMT, SampathH, et al.: Hepatic stearoyl-CoA desaturase-1 deficiency protects mice from carbohydrate-induced adiposity and hepatic steatosis. Cell Metab, 2007; 6:484–496.
43.
GuillouH, MartinP, JanS, et al.: Comparative effect of fenofibrate on hepatic desaturases in wild-type and peroxisome proliferator-activated receptor alpha-deficient mice. Lipids, 2002; 37:981–989.
44.
MillerCW, NtambiJM: Peroxisome proliferators induce mouse liver stearoyl-CoA desaturase 1 gene expression. Proc Natl Acad Sci U S A, 1996; 93:9443–9448.
WillsonTM, BrownPJ, SternbachDD, HenkeBR: The PPARs: From orphan receptors to drug discovery. J Med Chem, 2000; 43:527–550.
48.
BergerJ, MollerDE: The mechanisms of action of PPARs. Annu Rev Med, 2002; 53:409–435.
49.
ParkTS, PanekRL, MuellerSB, et al.: Inhibition of sphingomyelin synthesis reduces atherogenesis in apolipoprotein E-knockout mice. Circulation, 2004; 110:3465–3471.
50.
HollandWL, BrozinickJT, WangLP, et al.: Inhibition of ceramide synthesis ameliorates glucocorticoid-, saturated-fat-, and obesity-induced insulin resistance. Cell Metab, 2007; 5:167–179.
51.
ChaurasiaB, KaddaiVA, LancasterGI, et al.: Adipocyte ceramides regulate subcutaneous adipose browning, inflammation, and metabolism. Cell Metab, 2016; 24:820–834.
52.
KitadaY, KajitaK, TaguchiK, et al.: Blockade of sphingosine 1-phosphate receptor 2 signaling attenuates high-fat diet-induced adipocyte hypertrophy and systemic glucose intolerance in mice. Endocrinology, 2016; 157:1839–1851.
53.
MoonMH, JeongJK, LeeJH, et al.: Antiobesity activity of a sphingosine 1-phosphate analogue FTY720 observed in adipocytes and obese mouse model. Exp Mol Med, 2012; 44:603–614.
54.
HannunYA, ObeidLM: Principles of bioactive lipid signalling: Lessons from sphingolipids. Nat Rev Mol Cell Biol, 2008; 9:139–150.
55.
LeeSY, LeeHY, SongJH, et al.: Adipocyte-specific deficiency of de novo sphingolipid biosynthesis leads to lipodystrophy and insulin resistance. Diabetes, 2017; 66:2596–2609.
56.
ParhamKA, ZebolJR, TooleyKL, et al.: Sphingosine 1-phosphate is a ligand for peroxisome proliferator-activated receptor-gamma that regulates neoangiogenesis. FASEB J, 2015; 29:3638–3653.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
