Fatty acids do not pay the toll: effect of SFA and PUFA on human adipose tissue and mature adipocytes inflammation

Lipids in Health and Disease

Laurence Hoareau¹  Régis Roche¹  Maya Césari¹  Franck Festy¹  Lydie Gence¹  Karima Bencharif¹  Jibesh Kumar Padhan¹  Manoj Kumar Gunasekaran¹  Ravi Kumar Murumalla¹ 

[1] GEICO (Study Group on Chronic Inflammation and Obesity), Platform ‘Cyclotron Reunion Ocean Indien’ CYROI, 2, rue Maxime Rivière, Sainte-Clotilde, Reunion Island, 97 490, France

关键词: TLR;    Inflammation;    Human adipose tissue;    FFA;   

Others  :  1160112
DOI  :  10.1186/1476-511X-11-175

 received in 2012-05-31, accepted in 2012-12-18,  发布年份 2012

【 摘 要 】

Background

On the basis that high fat diet induces inflammation in adipose tissue, we wanted to test the effect of dietary saturated and polysunsaturated fatty acids on human adipose tissue and adipocytes inflammation. Moreover we wanted to determine if TLR2 and TLR4 are involved in this pathway.

Methods

Human adipose tissue and adipocytes primary cultures were treated with endotoxin-free BSA conjugated with SFA (lauric acid and palmitic acid - LA and PA) and PUFA (eicosapentaeneic acid, docosahexaenoic acid and oleic acid - EPA, DHA and OA) with or without LPS. Cytokines were then assayed by ELISA (TNF-alpha, IL-6 and MCP-1). In order to determine if TLR2 and TLR4 are activated by fatty acid (FA), we used HEK-Blue cells transfected by genes from TLR2 or TLR4 pathways associated with secreted alkaline phosphatase reporter gene.

Results

None of the FA tested in HEK-Blue cells were able to activate TLR2 or TLR4, which is concordant with the fact that after FA treatment, adipose tissue and adipocytes cytokines levels remain the same as controls. However, all the PUFA tested: DHA, EPA and to a lesser extent OA down-regulated TNF-alpha, IL-6 and MCP-1 secretion in human adipose tissue and adipocytes cultures.

Conclusions

This study first confirms that FA do not activate TLR2 and TLR4. Moreover by using endotoxin-free BSA, both SFA and PUFA tested were not proinflammatory in human adipose tissue and adipocytes model. More interestingly we showed that some PUFA exert an anti-inflammatory action in human adipose tissue and adipocytes model. These results are important since they clarify the relationship between dietary fatty acids and inflammation linked to obesity.

【 授权许可】

   
2012 Murumalla et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Guilherme A, Virbasius JV, Puri V, Czech MP: Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. Nat Rev Mol Cell Biol 2008, 9(5):367-377.
• [2]Aslan H, Altunkaynak BZ, Altunkaynak ME, Vuraler O, Kaplan S, Unal B: Effect of a high fat diet on quantitative features of adipocytes in the omentum: an experimental, stereological and ultrastructural study. Obes Surg 2006, 16(11):1526-1534.
• [3]Bullo M, Casas-Agustench P, Amigo-Correig P, Aranceta J, Salas-Salvado J: Inflammation, obesity and comorbidities: the role of diet. Public Health Nutr 2007, 10(10A):1164-1172.
• [4]Bays H, Mandarino L, DeFronzo RA: Role of the adipocyte, free fatty acids, and ectopic fat in pathogenesis of type 2 diabetes mellitus: peroxisomal proliferator-activated receptor agonists provide a rational therapeutic approach. J Clin Endocrinol Metab 2004, 89(2):463-478.
• [5]Suganami T, Tanimoto-Koyama K, Nishida J, Itoh M, Yuan X, Mizuarai S, Kotani H, Yamaoka S, Miyake K, Aoe S, et al.: Role of the Toll-like receptor 4/NF-kappaB pathway in saturated fatty acid-induced inflammatory changes in the interaction between adipocytes and macrophages. Arterioscler Thromb Vasc Biol 2007, 27(1):84-91.
• [6]Bunn RC, Cockrell GE, Ou Y, Thrailkill KM, Lumpkin CK Jr, Fowlkes JL: Palmitate and insulin synergistically induce IL-6 expression in human monocytes. Cardiovasc Diabetol 2010, 9:73. BioMed Central Full Text
• [7]Tsimikas S, Reaven PD: The role of dietary fatty acids in lipoprotein oxidation and atherosclerosis. Curr Opin Lipidol 1998, 9(4):301-307.
• [8]Hao W, Wong OY, Liu X, Lee P, Chen Y, Wong KKY: ω-3 fatty acids suppress inflammatory cytokine production by macrophages and hepatocytes. J Pediatr Surg 2010, 45(12):2412-2418.
• [9]Balvers MGJ, Verhoeckx KCM, Plastina P, Wortelboer HM, Meijerink J, Witkamp RF: Docosahexaenoic acid and eicosapentaenoic acid are converted by 3 T3-L1 adipocytes to N-acyl ethanolamines with anti-inflammatory properties. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids 2010, 1801(10):1107-1114.
• [10]Ibrahim A, Mbodji K, Hassan A, Aziz M, Boukhettala N, Coëffier M, Savoye G, Déchelotte P, Marion-Letellier R: Anti-inflammatory and anti-angiogenic effect of long chain n-3 polyunsaturated fatty acids in intestinal microvascular endothelium. Clin Nutr 2011, 30(5):678-687.
• [11]Mullen A, Loscher CE, Roche HM: Anti-inflammatory effects of EPA and DHA are dependent upon time and dose-response elements associated with LPS stimulation in THP-1-derived macrophages. J Nutr Biochem 2010, 21(5):444-450.
• [12]Ajuwon KM, Spurlock ME: Palmitate activates the NF-kappaB transcription factor and induces IL-6 and TNFalpha expression in 3 T3-L1 adipocytes. J Nutr 2005, 135(8):1841-1846.
• [13]Chang CF, Chau YP, Kung HN, Lu KS: The lipopolysaccharide-induced pro-inflammatory response in RAW264.7 cells is attenuated by an unsaturated fatty acid–bovine serum albumin complex and enhanced by a saturated fatty acid–bovine serum albumin complex. Inflamm Res 2011.
• [14]Harvey KA, Walker CL, Pavlina TM, Xu Z, Zaloga GP, Siddiqui RA: Long-chain saturated fatty acids induce pro-inflammatory responses and impact endothelial cell growth. Clin Nutr 2010, 29(4):492-500.
• [15]Hotamisligil GS: Inflammation and metabolic disorders. Nature 2006, 444(7121):860-867.
• [16]Shoelson SE, Lee J, Goldfine AB: Inflammation and insulin resistance. J Clin Invest 2006, 116(7):1793-1801.
• [17]Trayhurn P, Wood IS: Adipokines: inflammation and the pleiotropic role of white adipose tissue. Br J Nutr 2004, 92(3):347-355.
• [18]Bès-Houtmann S, Roche R, Hoareau L, Gonthier M-P, Festy F, Caillens H, Gasque P, Lefebvre d’Hellencourt C, Cesari M: Presence of functional TLR2 and TLR4 on human adipocytes. Histochem Cell Biol 2006, 127(2):131-137.
• [19]Samokhvalov V, Bilan PJ, Schertzer JD, Antonescu CN, Klip A: Palmitate- and lipopolysaccharide-activated macrophages evoke contrasting insulin responses in muscle cells. Am J Physiol Endocrinol Metab 2009, 296(1):E37-E46.
• [20]Haversen L, Danielsson KN, Fogelstrand L, Wiklund O: Induction of proinflammatory cytokines by long-chain saturated fatty acids in human macrophages. Atherosclerosis 2009, 202(2):382-393.
• [21]Weatherill AR, Lee JY, Zhao L, Lemay DG, Youn HS, Hwang DH: Saturated and polyunsaturated fatty acids reciprocally modulate dendritic cell functions mediated through TLR4. J Immunol 2005, 174(9):5390-5397.
• [22]Jin MS, Lee JO: Structures of the toll-like receptor family and its ligand complexes. Immunity 2008, 29(2):182-191.
• [23]Erridge C, Samani NJ: Saturated fatty acids Do Not directly stimulate toll-like receptor signaling. Arterioscler Thromb Vasc Biol 2009, 29(11):1944-1949.
• [24]Park JS, Lee EJ, Lee JC, Kim WK, Kim HS: Anti-inflammatory effects of short chain fatty acids in IFN-gamma-stimulated RAW 264.7 murine macrophage cells: involvement of NF-kappaB and ERK signaling pathways. Int Immunopharmacol 2007, 7(1):70-77.
• [25]Davis JE, Gabler NK, Walker-Daniels J, Spurlock ME: Tlr-4 deficiency selectively protects against obesity induced by diets high in saturated fat. Obesity (Silver Spring) 2008, 16(6):1248-1255.
• [26]Coenen KR, Gruen ML, Lee-Young RS, Puglisi MJ, Wasserman DH, Hasty AH: Impact of macrophage toll-like receptor 4 deficiency on macrophage infiltration into adipose tissue and the artery wall in mice. Diabetologia 2009, 52(2):318-328.
• [27]Mullick AE: Modulation of atherosclerosis in mice by toll-like receptor 2. J Clin Invest 2005, 115(11):3149-3156.
• [28]Poggi M, Bastelica D, Gual P, Iglesias MA, Gremeaux T, Knauf C, Peiretti F, Verdier M, Juhan-Vague I, Tanti JF, et al.: C3H/HeJ mice carrying a toll-like receptor 4 mutation are protected against the development of insulin resistance in white adipose tissue in response to a high-fat diet. Diabetologia 2007, 50(6):1267-1276.
• [29]Radin MS, Sinha S, Bhatt BA, Dedousis N, O’Doherty RM: Inhibition or deletion of the lipopolysaccharide receptor toll-like receptor-4 confers partial protection against lipid-induced insulin resistance in rodent skeletal muscle. Diabetologia 2007, 51(2):336-346.
• [30]Lee JY: Saturated fatty acid activates but polyunsaturated fatty acid inhibits toll-like receptor 2 dimerized with toll-like receptor 6 or 1. J Biol Chem 2004, 279(17):16971-16979.
• [31]Lee JY, Sohn KH, Rhee SH, Hwang D: Saturated fatty acids, but not unsaturated fatty acids, induce the expression of cyclooxygenase-2 mediated through toll-like receptor 4. J Biol Chem 2001, 276(20):16683-16689.
• [32]Feng D, Tang Y, Kwon H, Zong H, Hawkins M, Kitsis RN, Pessin JE: High-fat diet-induced adipocyte cell death occurs through a cyclophilin D intrinsic signaling pathway independent of adipose tissue inflammation. Diabetes 2011, 60(8):2134-2143.
• [33]Han CY, Kargi AY, Omer M, Chan CK, Wabitsch M, O’Brien KD, Wight TN, Chait A: Differential effect of saturated and unsaturated free fatty acids on the generation of monocyte adhesion and chemotactic factors by adipocytes: dissociation of adipocyte hypertrophy from inflammation. Diabetes 2009, 59(2):386-396.
• [34]Weigert C, Brodbeck K, Staiger H, Kausch C, Machicao F, Haring HU, Schleicher ED: Palmitate, but not unsaturated fatty acids, induces the expression of interleukin-6 in human myotubes through proteasome-dependent activation of nuclear factor-kappaB. J Biol Chem 2004, 279(23):23942-23952.
• [35]Li W, Sama AE, Wang H: Role of HMGB1 in cardiovascular diseases. Curr Opin Pharmacol 2006, 6(2):130-135.
• [36]Schwartz EA, Zhang WY, Karnik SK, Borwege S, Anand VR, Laine PS, Su Y, Reaven PD: Nutrient modification of the innate immune response: a novel mechanism by which saturated fatty acids greatly amplify monocyte inflammation. Arterioscler Thromb Vasc Biol 2010, 30(4):802-808.
• [37]Ismail NA, Ragab S, El Dayem SM, Elbaky AA, Salah N, Hamed M, Assal H, Koura H: Fetuin-A levels in obesity: differences in relation to metabolic syndrome and correlation with clinical and laboratory variables. Arch Med Sci 2012, 8(5):826-833.
• [38]Erdmann J, Salmhofer H, Knauss A, Mayr M, Wagenpfeil S, Sypchenko O, Luppa P, Schusdziarra V: Relationship of fetuin-A levels to weight-dependent insulin resistance and type 2 diabetes mellitus. Regul Pept 2012, 178(1–3):6-10.
• [39]Singh M, Sharma PK, Garg VK, Mondal SC, Singh AK, Kumar N: Role of fetuin-A in atherosclerosis associated with diabetic patients. J Pharm Pharmacol 2012, 64(12):1703-1708.
• [40]Pal D, Dasgupta S, Kundu R, Maitra S, Das G, Mukhopadhyay S, Ray S, Majumdar SS, Bhattacharya S: Fetuin-A acts as an endogenous ligand of TLR4 to promote lipid-induced insulin resistance. Nat Med 2012.
• [41]Skuladottir IH, Petursdottir DH, Hardardottir I: The effects of omega-3 polyunsaturated fatty acids on TNF-alpha and IL-10 secretion by murine peritoneal cells in vitro. Lipids 2007, 42(8):699-706.
• [42]Gotoh N, Nagao K, Onoda S, Shirouchi B, Furuya K, Nagai T, Mizobe H, Ichioka K, Watanabe H, Yanagita T, et al.: Effects of three different highly purified n-3 series highly unsaturated fatty acids on lipid metabolism in C57BL/KsJ-db/dbMice. J Agric Food Chem 2009, 57(22):11047-11054.
• [43]Lu J, Jilling T, Li D, Caplan MS: Polyunsaturated fatty acid supplementation alters proinflammatory gene expression and reduces the incidence of necrotizing enterocolitis in a neonatal rat model. Pediatr Res 2007, 61(4):427-432.
• [44]Panda SK, Kumar S, Tupperwar NC, Vaidya T, George A, Rath S, Bal V, Ravindran B: Chitohexaose activates macrophages by alternate pathway through TLR4 and blocks endotoxemia. PLoS Pathog 2012, 8(5):e1002717.
• [45]Wang M, Chen Y, Zhang Y, Zhang L, Lu X, Chen Z: Mannan-binding lectin directly interacts with toll-like receptor 4 and suppresses lipopolysaccharide-induced inflammatory cytokine secretion from THP-1 cells. Cell Mol Immunol 2011, 8(3):265-275.
• [46]Wong SW, Kwon MJ, Choi AM, Kim HP, Nakahira K, Hwang DH: Fatty acids modulate toll-like receptor 4 activation through regulation of receptor dimerization and recruitment into lipid rafts in a reactive oxygen species-dependent manner. J Biol Chem 2009, 284(40):27384-27392.
• [47]Hoareau L, Bencharif K, Rondeau P, Murumalla R, Ravanan P, Tallet F, Delarue P, Cesari M, Roche R, Festy F: Signaling pathways involved in LPS induced TNFalpha production in human adipocytes. J Inflamm (Lond) 2010, 7:1. BioMed Central Full Text
• [48]Nair S, Lee YH, Rousseau E, Cam M, Tataranni PA, Baier LJ, Bogardus C, Permana PA: Increased expression of inflammation-related genes in cultured preadipocytes/stromal vascular cells from obese compared with non-obese Pima Indians. Diabetologia 2005, 48(9):1784-1788.
• [49]Hoareau L, Buyse M, Festy F, Ravanan P, Gonthier MP, Matias I, Petrosino S, Tallet F, d’Hellencourt CL, Cesari M, et al.: Anti-inflammatory effect of palmitoylethanolamide on human adipocytes. Obesity (Silver Spring) 2009, 17(3):431-438.
• [50]Fang S, Suh JM, Atkins AR, Hong SH, Leblanc M, Nofsinger RR, Yu RT, Downes M, Evans RM: Corepressor SMRT promotes oxidative phosphorylation in adipose tissue and protects against diet-induced obesity and insulin resistance. Proc Natl Acad Sci U S A 2011, 108(8):3412-3417.
• [51]Ye Q, Chen Y, Lei H, Liu Q, Moorhead JF, Varghese Z, Ruan XZ: Inflammatory stress increases unmodified LDL uptake via LDL receptor: an alternative pathway for macrophage foam-cell formation. Inflamm Res 2009, 58(11):809-818.
• [52]Eder K, Guan H, Sung HY, Francis SE, Crossman DC, Kiss-Toth E: LDL uptake by monocytes in response to inflammation is MAPK dependent but independent of tribbles protein expression. Immunol Lett 2008, 116(2):178-183.