We previously described a negative effect of xanthohumol (XN) upon placentation-related processes. We aimed to better characterize this effect by investigating the effect of XN upon the uptake of arachidonic acid (ARA), a crucial nutrient during pregnancy, by the HTR-8/SVneo human first-trimester extravillous trophoblast cell line and its relationship with the negative effect of XN upon placentation-related processes. Uptake of 14C-ARA (100 nM) was time dependent and inhibited by short-term (26 minutes) or long-term (24 hours) exposure to XN. Xanthohumol (24 hours; 5 µM) behaved as an uncompetitive inhibitor of 14C-ARA uptake; the mammalian target of rapamycin, tyrosine kinases, and c-Jun N-terminal kinases intracellular pathways were involved in this effect; and it markedly reduced long-chain acyl-CoA synthetase 1 messenger RNA levels. Moreover, the effects of XN (24 hours; 5 µM) upon cell proliferation, culture growth, migration, viability, and apoptosis index were prevented by high extracellular ARA but not by the peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist rosiglitazone. We thus conclude that ARA is an essential nutrient regulating cell viability, proliferation, culture growth, migration, and apoptosis of HTR-8/SVneo cells and that the deleterious effects of XN involve inhibition of ARA cellular uptake but appears to be independent of PPAR-γ activation.
JamesJLCarterAMChamleyLW. Human placentation from nidation to 5 weeks of gestation. Part I: what do we know about formative placental development following implantation?Placenta. 2012;33(5):327–334.
2.
JiLBrkicJLiuMFuGPengCWangYL. Placental trophoblast cell differentiation: physiological regulation and pathological relevance to preeclampsia. Mol Aspects Med. 2013;34(5):981–1023.
3.
Dutta-RoyAK. Transport mechanisms for long-chain polyunsaturated fatty acids in the human placenta. Am J Clin Nutr. 2000;71(suppl 1):315S–322S.
4.
InnisSM. Fatty acids and early human development. Early Hum Dev. 2007;83(12):761–766.
5.
CunninghamPMcDermottL. Long chain PUFA transport in human term placenta. J Nutr. 2009;139(4):636–639.
6.
KoletzkoBLienEAgostoniC. The roles of long-chain polyunsaturated fatty acids in pregnancy, lactation and infancy: review of current knowledge and consensus recommendations. J Perinat Med. 2008;36(1):5–14.
7.
LagerSPowellTL. Regulation of nutrient transport across the placenta. J Pregnancy. 2012;2012:179827.
8.
AraujoJRRamalhoCCorreia-BrancoA. A parallel increase in placental oxidative stress and antioxidant defenses occurs in pre-gestational type 1 but not gestational diabetes. Placenta. 2013;34(11):1095–1098.
9.
TobinKAJohnsenGMStaffACDuttaroyAK. Long-chain polyunsaturated fatty acid transport across human placental choriocarcinoma (BeWo) cells. Placenta. 2009;30(1):41–47.
10.
BasakSDasMKDuttaroyAK. Fatty acid-induced angiogenesis in first trimester placental trophoblast cells: possible roles of cellular fatty acid-binding proteins. Life Sci. 2013;93(21):755–762.
11.
StevensJFPageJE. Xanthohumol and related prenylflavonoids from hops and beer: to your good health!Phytochemistry. 2004;65(10):1317–1330.
12.
Correia-BrancoAAzevedoCFAraujoJR. Xanthohumol impairs glucose uptake by a human first-trimester extravillous trophoblast cell line (HTR-8/SVneo cells) and impacts the process of placentation. Mol Hum Reprod. 2015;21(10):803–815.
13.
AraujoJRGoncalvesPMartelF. Modulation of glucose uptake in a human choriocarcinoma cell line (BeWo) by dietary bioactive compounds and drugs of abuse. J Biochem. 2008;144(2):177–186.
14.
BradfordMM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–254.
15.
BergmeyerHUBerntE. Lactate dehydrogenase. In: BergmeyerHU, editor. Methods of Enzymatic Analysis, 2nd ed., New York: Academic Press, 1974;II:574–9.
16.
NegraoRCostaRDuarteDGomesTTAzevedoISoaresR. Different effects of catechin on angiogenesis and inflammation depending on VEGF levels. J Nutr Biochem. 2013;24(2):435–444.
17.
Reinhart-KingCA. Endothelial cell adhesion and migration. Methods Enzymol. 2008;443:45–64.
18.
SchmittgenTDLivakKJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008;3(6):1101–1108.
19.
MuzykaARMNovikovAMoskalenkoEVysotskyAVolokhV. Graphpad Prism From Windows. Version 5.03. 2009.
20.
PohlJRingAEhehaltRHerrmannTStremmelW. New concepts of cellular fatty acid uptake: role of fatty acid transport proteins and of caveolae. Proc Nutr Soc. 2004;63(2):259–262.
21.
KaleANaphadeNSapkaleS. Reduced folic acid, vitamin B12 and docosahexaenoic acid and increased homocysteine and cortisol in never-medicated schizophrenia patients: implications for altered one-carbon metabolism. Psychiatry Res. 2010;175(1-2):47–53.
22.
QiuAJansenMSakarisA. Identification of an intestinal folate transporter and the molecular basis for hereditary folate malabsorption. Cell. 2006;127(5):917–928.
23.
SchaiffWTBildiriciICheongMChernPLNelsonDMSadovskyY. Peroxisome proliferator-activated receptor-gamma and retinoid × receptor signaling regulate fatty acid uptake by primary human placental trophoblasts. J Clin Endocrinol Metab. 2005;90(7):4267–4275.
24.
FournierTPavanLTarradeA. The role of PPAR-gamma/RXR-alpha heterodimers in the regulation of human trophoblast invasion. Ann N Y Acad Sci. 2002;973:26–30.
25.
RauwelBMariameBMartinH. Activation of peroxisome proliferator-activated receptor gamma by human cytomegalovirus for de novo replication impairs migration and invasiveness of cytotrophoblasts from early placentas. J Virol. 2010;84(6):2946–2954.
26.
BilbanMHaslingerPPrastJ. Identification of novel trophoblast invasion-related genes: heme oxygenase-1 controls motility via peroxisome proliferator-activated receptor gamma. Endocrinology. 2009;150(2):1000–1013.
27.
FisherSJ. The placental problem: linking abnormal cytotrophoblast differentiation to the maternal symptoms of preeclampsia. Reprod Biol Endocrinol. 2004;2:53.
28.
GrahamCHHawleyTSHawleyRG. Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 1993;206(2):204–211.
29.
JohnsenGMWeedon-FekjaerMSTobinKAStaffACDuttaroyAK. Long-chain polyunsaturated fatty acids stimulate cellular fatty acid uptake in human placental choriocarcinoma (BeWo) cells. Placenta. 2009;30(12):1037–1044.
30.
BonenAChabowskiALuikenJJGlatzJF. Is membrane transport of FFA mediated by lipid, protein, or both? Mechanisms and regulation of protein-mediated cellular fatty acid uptake: molecular, biochemical, and physiological evidence. Physiology (Bethesda). 2007;22:15–29.
31.
DoegeHStahlA. Protein-mediated fatty acid uptake: novel insights from in vivo models. Physiology (Bethesda). 2006;21:259–268.
32.
CampbellFMGordonMJDutta-RoyAK. Preferential uptake of long chain polyunsaturated fatty acids by isolated human placental membranes. Mol Cell Biochem. 1996;155(1):77–83.
33.
QiuQYangMTsangBKGruslinA. Both mitogen-activated protein kinase and phosphatidylinositol 3-kinase signalling are required in epidermal growth factor-induced human trophoblast migration. Mol Hum Reprod. 2004;10(9):677–684.
34.
GangloffYGMuellerMDannSG. Disruption of the mouse mTOR gene leads to early postimplantation lethality and prohibits embryonic stem cell development. Mol Cell Biol. 2004;24(21):9508–9516.
35.
MurakamiMIchisakaTMaedaM. mTOR is essential for growth and proliferation in early mouse embryos and embryonic stem cells. Mol Cell Biol. 2004;24(15):6710–6718.
36.
WenHYAbbasiSKellemsREXiaY. mTOR: a placental growth signaling sensor. Placenta. 2005;26(suppl A):S63–S69.
37.
LiuLWangYShenC. Benzo(a)pyrene inhibits migration and invasion of extravillous trophoblast HTR-8/SVneo cells via activation of the ERK and JNK pathway. J Appl Toxicol. 2016;36(7):946–955.
38.
DimasuayKGBoeufPPowellTLJanssonT. Placental responses to changes in the maternal environment determine fetal growth. Front Physiol. 2016;7:12.
39.
JanssonTAyeILGoberdhanDC. The emerging role of mTORC1 signaling in placental nutrient-sensing. Placenta. 2012;33(suppl 2):e23–e29.
ManachCWilliamsonGMorandCScalbertARémésyC. Bioavailability and bioefficacy of polyphenols in humans. I. Review of 97 bioavailability studies. Am J Clin Nutr. 2005;81(suppl 1):230S–242S.
42.
FormanBMChenJEvansRM. Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors alpha and delta. Proc Natl Acad Sci U S A. 1997;94(9):4312–4317.
XuYWangQCookTJKnippGT. Effect of placental fatty acid metabolism and regulation by peroxisome proliferator activated receptor on pregnancy and fetal outcomes. J Pharm Sci. 2007;96(10):2582–2606.
45.
DesvergneBMichalikLWahliW. Transcriptional regulation of metabolism. Physiol Rev. 2006;86(2):465–514.
46.
CapobiancoEJawerbaumARomaniniMC. 15-Deoxy-Delta12,14-prostaglandin J2 and peroxisome proliferator-activated receptor gamma (PPARgamma) levels in term placental tissues from control and diabetic rats: modulatory effects of a PPARgamma agonist on nitridergic and lipid placental metabolism. Reprod Fertil Dev. 2005;17(4):423–433.
47.
SchaiffWTKnappFFJrBarakYBiron-ShentalTNelsonDMSadovskyY. Ligand-activated peroxisome proliferator activated receptor gamma alters placental morphology and placental fatty acid uptake in mice. Endocrinology. 2007;148(8):3625–3634.
48.
BarakYNelsonMCOngES. PPAR gamma is required for placental, cardiac, and adipose tissue development. Mol Cell. 1999;4(4):585–595.
49.
TarradeASchoonjansKPavanL. PPARgamma/RXRalpha heterodimers control human trophoblast invasion. J Clin Endocrinol Metab. 2001;86(10):5017–5024.
50.
McCarthyFPDrewloSKingdomJJohnsEJWalshSKKennyLC. Peroxisome proliferator-activated receptor-gamma as a potential therapeutic target in the treatment of preeclampsia. Hypertension. 2011;58(2):280–286.
51.
MontellanoPR. The mechanism of heme oxygenase. Curr Opin Chem Biol. 2000;4(2):221–227.
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