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A20 reduces lipid storage and inflammation in hypertrophic adipocytes via p38 and Akt signaling

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Abstract

Adipose tissue plays a vital role in the development of obesity and related disorders. Our previous study showed that A20, an ubiquitin-editing enzyme with anti-inflammation function, attenuated free fatty acids (FFAs)-induced lipid accumulation in nonalcoholic steatohepatitis. Here, we investigated A20 expression in adipose tissue of obese individuals and its effects on 3T3-L1 lipogenesis as well as the likely mechanisms underlying this process. By re-annotation of raw microarray data downloaded from Gene Expression Omnibus, we found that obese individuals showed significantly higher A20 mRNA levels in adipocytes. In vitro, A20 inhibited MCP-1 and IL-6 secretion in adipocytes. Forced expression of A20 resulted in decreased expression of key markers of lipogenesis and adipogenesis, such as sterol regulatory element binding protein 1c (SREBP-1c) and adipogenesis (aP2), leading to less lipids accumulation in differentiated 3T3-L1 cells. This process was concomitant with attenuated activation of p38 and Akt signaling. Our results suggest that A20 may have therapeutic potential for obesity and related diseases. The mechanisms involved the suppression of lipid storage and inflammation in adipocytes.

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Abbreviations

MCD:

Methionine choline-deficient

NAFLD:

Nonalcoholic fatty liver disease

NASH:

Nonalcoholic steatohepatitis

FFAs:

Free fatty acids

TNFAIP3:

TNF-α-induced protein 3

SREBP-1c:

Sterol regulatory element binding protein1c

ACC1:

Acetyl-CoA carboxylase 1

FAS:

Fatty acid synthase

OA:

Oleic acid

PA:

Palmitic acid

IL-6:

Interleukin-6

Dgat2:

Diacylglycerol O-acyltransferase-2

Scd-1:

Stearoyl-CoA desaturase

References

  1. Guilherme A, Virbasius JV, Puri V, Czech MP (2008) Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. Nat Rev Mol Cell Biol 9:367–377. doi:10.1038/nrm2391

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Abenavoli L, DIR L, Guzzi PH, Pellicano R, Milic N, DEL A (2015) Non-alcoholic fatty liver disease severity, central fat mass and adinopectin: a close relationship. Clujul Med 88:489–493. doi:10.15386/cjmed-595

    Article  PubMed  PubMed Central  Google Scholar 

  3. Jung UJ, Choi MS (2014) Obesity and its metabolic complications: the role of adipokines and the relationship between obesity, inflammation, insulin resistance, dyslipidemia and nonalcoholic fatty liver disease. Int J Mol Sci 15:6184–6223. doi:10.3390/ijms15046184

    Article  PubMed  PubMed Central  Google Scholar 

  4. Ajuwon KM, Spurlock ME (2005) Palmitate activates the NF-kappaB transcription factor and induces IL-6 and TNFalpha expression in 3T3-L1 adipocytes. J Nutr 135:1841–1846

    CAS  PubMed  Google Scholar 

  5. Boren J, Taskinen MR, Olofsson SO, Levin M (2013) Ectopic lipid storage and insulin resistance: a harmful relationship. J Intern Med 274:25–40. doi:10.1111/joim.12071

    Article  CAS  PubMed  Google Scholar 

  6. Ai L, Xu Q, Wu C, Wang X, Chen Z, Su D, Jiang X, Xu A, Lin Q, Fan Z (2015) A20 Attenuates FFAs-induced lipid accumulation in nonalcoholic steatohepatitis. Int J Biol Sci 11:1436–1446. doi:10.7150/ijbs.13371

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Verstrepen L, Verhelst K, van Loo G, Carpentier I, Ley SC, Beyaert R (2010) Expression, biological activities and mechanisms of action of A20 (TNFAIP3). Biochem Pharmacol 80:2009–2020. doi:10.1016/j.bcp.2010.06.044

    Article  CAS  PubMed  Google Scholar 

  8. Stewart TP, Kim HY, Saxton AM, Kim JH (2010) Genetic and genomic analysis of hyperlipidemia, obesity and diabetes using (C57BL/6 J x TALLYHO/JngJ) F2 mice. BMC Genom 11:713. doi:10.1186/1471-2164-11-713

    Article  CAS  Google Scholar 

  9. Lee YH, Nair S, Rousseau E, Allison DB, Page GP, Tataranni PA, Bogardus C, Permana PA (2005) Microarray profiling of isolated abdominal subcutaneous adipocytes from obese vs non-obese Pima Indians: increased expression of inflammation-related genes. Diabetologia 48:1776–1783. doi:10.1007/s00125-005-1867-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Onate B, Vilahur G, Camino-Lopez S, Diez-Caballero A, Ballesta-Lopez C, Ybarra J, Moscatiello F, Herrero J, Badimon L (2013) Stem cells isolated from adipose tissue of obese patients show changes in their transcriptomic profile that indicate loss in stem cellness and increased commitment to an adipocyte-like phenotype. BMC Genom 14:625. doi:10.1186/1471-2164-14-625

    Article  CAS  Google Scholar 

  11. Gesta S, Bluher M, Yamamoto Y, Norris AW, Berndt J, Kralisch S, Boucher J, Lewis C, Kahn CR (2006) Evidence for a role of developmental genes in the origin of obesity and body fat distribution. Proc Natl Acad Sci USA 103:6676–6681. doi:10.1073/pnas.0601752103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Xu AT, Li Y, Zhao D, Shen J, Xu XT, Qiao YQ, Zhu MM, Wang TR, Cui Y, Ai LY, Ran ZH (2015) High suppressor of cytokine signaling-3 expression impairs stat3-dependent protective effects of interleukin-22 in ulcerative colitis in remission. Inflamm Bowel Dis 21:241–250. doi:10.1097/MIB.0000000000000267

    Article  PubMed  Google Scholar 

  13. Chen ZF, Ai LY, Wang JL, Ren LL, Yu YN, Xu J, Chen HY, Yu J, Li M, Qin WX, Ma X, Shen N, Chen YX, Hong J, Fang JY (2015) Probiotics Clostridium butyricum and Bacillus subtilis ameliorate intestinal tumorigenesis. Future Microbiol 10:1433–1445. doi:10.2217/fmb.15.66

    Article  CAS  PubMed  Google Scholar 

  14. Chawla A, Nguyen KD, Goh YP (2011) Macrophage-mediated inflammation in metabolic disease. Nat Rev Immunol 11:738–749. doi:10.1038/nri3071

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Lumeng CN, Saltiel AR (2011) Inflammatory links between obesity and metabolic disease. J Clin Invest 121:2111–2117. doi:10.1172/jci57132

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. He Z, Li M, Zheng D, Chen Q, Liu W, Feng L (2015) Adipose tissue hypoxia and low-grade inflammation: a possible mechanism for ethanol-related glucose intolerance? Br J Nutr 113:1355–1364. doi:10.1017/S000711451500077X

    Article  CAS  PubMed  Google Scholar 

  17. Priceman SJ, Kujawski M, Shen S, Cherryholmes GA, Lee H, Zhang C, Kruper L, Mortimer J, Jove R, Riggs AD, Yu H (2013) Regulation of adipose tissue T cell subsets by Stat3 is crucial for diet-induced obesity and insulin resistance. Proc Natl Acad Sci USA 110:13079–13084. doi:10.1073/pnas.1311557110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Wertz IE, Newton K, Seshasayee D, Kusam S, Lam C, Zhang J, Popovych N, Helgason E, Schoeffler A, Jeet S, Ramamoorthi N, Kategaya L, Newman RJ, Horikawa K, Dugger D, Sandoval W, Mukund S, Zindal A, Martin F, Quan C, Tom J, Fairbrother WJ, Townsend M, Warming S, DeVoss J, Liu J, Dueber E, Caplazi P, Lee WP, Goodnow CC, Balazs M, Yu K, Kolumam G, Dixit VM (2015) Phosphorylation and linear ubiquitin direct A20 inhibition of inflammation. Nature 528:370–375. doi:10.1038/nature16165

    Article  CAS  PubMed  Google Scholar 

  19. Arvelo MB, Cooper JT, Longo C, Daniel S, Grey ST, Mahiou J, Czismadia E, Abu-Jawdeh G, Ferran C (2002) A20 protects mice from D-galactosamine/lipopolysaccharide acute toxic lethal hepatitis. Hepatology 35:535–543. doi:10.1053/jhep.2002.31309

    Article  CAS  PubMed  Google Scholar 

  20. Hymowitz SG, Wertz IE (2010) A20: from ubiquitin editing to tumour suppression. Nat Rev Cancer 10:332–341. doi:10.1038/nrc2775

    Article  CAS  PubMed  Google Scholar 

  21. Duong BH, Onizawa M, Oses-Prieto JA, Advincula R, Burlingame A, Malynn BA, Ma A (2015) A20 restricts ubiquitination of pro-interleukin-1beta protein complexes and suppresses NLRP3 inflammasome activity. Immunity 42:55–67. doi:10.1016/j.immuni.2014.12.031

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Cooper JT, Stroka DM, Brostjan C, Palmetshofer A, Bach FH, Ferran C (1996) A20 blocks endothelial cell activation through a NF-kappaB-dependent mechanism. J Biol Chem 271:18068–18073

    Article  CAS  PubMed  Google Scholar 

  23. Engelman JA, Lisanti MP, Scherer PE (1998) Specific inhibitors of p38 mitogen-activated protein kinase block 3T3-L1 adipogenesis. J Biol Chem 273:32111–32120

    Article  CAS  PubMed  Google Scholar 

  24. Farmer SR (2006) Transcriptional control of adipocyte formation. Cell Metab 4:263–273. doi:10.1016/j.cmet.2006.07.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Bohgaki M, Tsukiyama T, Nakajima A, Maruyama S, Watanabe M, Koike T, Hatakeyama S (2008) Involvement of Ymer in suppression of NF-kappaB activation by regulated interaction with lysine-63-linked polyubiquitin chain. Biochim Biophys Acta 1783:826–837. doi:10.1016/j.bbamcr.2007.09.006

    Article  CAS  PubMed  Google Scholar 

  26. Lee EG, Boone DL, Chai S, Libby SL, Chien M, Lodolce JP, Ma A (2000) Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice. Science 289:2350–2354

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Chen NK, Chong TW, Loh HL, Lim KH, Gan VH, Wang M, Kon OL (2013) Negative regulatory responses to metabolically triggered inflammation impair renal epithelial immunity in diabetes mellitus. J Mol Med (Berl) 91:587–598. doi:10.1007/s00109-012-0969-x

    Article  CAS  Google Scholar 

  28. Feldstein AE, Werneburg NW, Canbay A, Guicciardi ME, Bronk SF, Rydzewski R, Burgart LJ, Gores GJ (2004) Free fatty acids promote hepatic lipotoxicity by stimulating TNF-alpha expression via a lysosomal pathway. Hepatology 40:185–194. doi:10.1002/hep.20283

    Article  CAS  PubMed  Google Scholar 

  29. Opipari AW Jr, Boguski MS, Dixit VM (1990) The A20 cDNA induced by tumor necrosis factor alpha encodes a novel type of zinc finger protein. J Biol Chem 265:14705–14708

    CAS  PubMed  Google Scholar 

  30. Sarma V, Lin Z, Clark L, Rust BM, Tewari M, Noelle RJ, Dixit VM (1995) Activation of the B-cell surface receptor CD40 induces A20, a novel zinc finger protein that inhibits apoptosis. J Biol Chem 270:12343–12346

    Article  CAS  PubMed  Google Scholar 

  31. Grey ST, Arvelo MB, Hasenkamp W, Bach FH, Ferran C (1999) A20 inhibits cytokine-induced apoptosis and nuclear factor kappaB-dependent gene activation in islets. J Exp Med 190:1135–1146

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Dorronsoro A, Lang V, Jakobsson E, Ferrin I, Salcedo JM, Fernandez-Rueda J, Fechter K, Rodriguez MS, Trigueros C (2013) Identification of the NF-kappaB inhibitor A20 as a key regulator for human adipogenesis. Cell Death Dis 4:e972. doi:10.1038/cddis.2013.494

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Coornaert B, Baens M, Heyninck K, Bekaert T, Haegman M, Staal J, Sun L, Chen ZJ, Marynen P, Beyaert R (2008) T cell antigen receptor stimulation induces MALT1 paracaspase-mediated cleavage of the NF-kappaB inhibitor A20. Nat Immunol 9:263–271. doi:10.1038/ni1561

    Article  CAS  PubMed  Google Scholar 

  34. Duwel M, Welteke V, Oeckinghaus A, Baens M, Kloo B, Ferch U, Darnay BG, Ruland J, Marynen P, Krappmann D (2009) A20 negatively regulates T cell receptor signaling to NF-kappaB by cleaving Malt1 ubiquitin chains. J Immunol 182:7718–7728. doi:10.4049/jimmunol.0803313

    Article  PubMed  Google Scholar 

  35. Longo CR, Patel VI, Shrikhande GV, Scali ST, Csizmadia E, Daniel S, Sun DW, Grey ST, Arvelo MB, Ferran C (2005) A20 protects mice from lethal radical hepatectomy by promoting hepatocyte proliferation via a p21waf1-dependent mechanism. Hepatology 42:156–164. doi:10.1002/hep.20741

    Article  CAS  PubMed  Google Scholar 

  36. Berndt J, Kovacs P, Ruschke K, Kloting N, Fasshauer M, Schon MR, Korner A, Stumvoll M, Bluher M (2007) Fatty acid synthase gene expression in human adipose tissue: association with obesity and type 2 diabetes. Diabetologia 50:1472–1480. doi:10.1007/s00125-007-0689-x

    Article  CAS  PubMed  Google Scholar 

  37. Loftus TM, Jaworsky DE, Frehywot GL, Townsend CA, Ronnett GV, Lane MD, Kuhajda FP (2000) Reduced food intake and body weight in mice treated with fatty acid synthase inhibitors. Science 288:2379–2381

    Article  CAS  PubMed  Google Scholar 

  38. Wu M, Singh SB, Wang J, Chung CC, Salituro G, Karanam BV, Lee SH, Powles M, Ellsworth KP, Lassman ME, Miller C, Myers RW, Tota MR, Zhang BB, Li C (2011) Antidiabetic and antisteatotic effects of the selective fatty acid synthase (FAS) inhibitor platensimycin in mouse models of diabetes. Proc Natl Acad Sci USA 108:5378–5383. doi:10.1073/pnas.1002588108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Smith SJ, Cases S, Jensen DR, Chen HC, Sande E, Tow B, Sanan DA, Raber J, Eckel RH, Farese RV Jr (2000) Obesity resistance and multiple mechanisms of triglyceride synthesis in mice lacking Dgat. Nat Genet 25:87–90. doi:10.1038/75651

    Article  CAS  PubMed  Google Scholar 

  40. Zhang Z, Shu G, Zhu X, Guo J, Cai H, Wang S, Wang L, Gao P, Xi Q, Zhang Y, Yuan L, Jiang Q (2014) Effect of diacylglycerol acyltransferase 2 overexpression in 3T3-L1 is associated to an increase in mono-unsaturated fatty acid accumulation. J Anim Sci Biotechnol 5:29. doi:10.1186/2049-1891-5-29

    Article  PubMed  PubMed Central  Google Scholar 

  41. Makowski L, Hotamisligil GS (2004) Fatty acid binding proteins—the evolutionary crossroads of inflammatory and metabolic responses. J Nutr 134:2464S–2468S

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Xu A, Wang Y, Xu JY, Stejskal D, Tam S, Zhang J, Wat NM, Wong WK, Lam KS (2006) Adipocyte fatty acid-binding protein is a plasma biomarker closely associated with obesity and metabolic syndrome. Clin Chem 52:405–413. doi:10.1373/clinchem.2005.062463

    Article  CAS  PubMed  Google Scholar 

  43. Maeda K (2007) Role of adiponectin and adipocyte fatty acid binding protein in the metabolic syndrome. Diabetes Res Clin Pract 77(Suppl 1):S17–S22. doi:10.1016/j.diabres.2007.01.028

    Article  CAS  PubMed  Google Scholar 

  44. Kilroy G, Carter LE, Newman S, Burk DH, Manuel J, Moller A, Bowtell DD, Mynatt RL, Ghosh S, Floyd ZE (2015) The ubiquitin ligase Siah2 regulates obesity-induced adipose tissue inflammation. Obesity (Silver Spring) 23:2223–2232. doi:10.1002/oby.21220

    Article  CAS  Google Scholar 

  45. Aouadi M, Jager J, Laurent K, Gonzalez T, Cormont M, Binetruy B, Le Marchand-Brustel Y, Tanti JF, Bost F (2007) p38MAP Kinase activity is required for human primary adipocyte differentiation. FEBS Lett 581:5591–5596. doi:10.1016/j.febslet.2007.10.064

    Article  CAS  PubMed  Google Scholar 

  46. Ji J, Zhu J, Hu X, Wang T, Zhang X, Hou AJ, Wang H (2015) (2S)-7,4′-dihydroxy-8-prenylflavan stimulates adipogenesis and glucose uptake through p38MAPK pathway in 3T3-L1 cells. Biochem Biophys Res Commun 460:578–582. doi:10.1016/j.bbrc.2015.03.072

    Article  CAS  PubMed  Google Scholar 

  47. Wang S, Xu Q, Shu G, Wang L, Gao P, Xi Q, Zhang Y, Jiang Q, Zhu X (2015) N-oleoyl glycine, a lipoamino acid, stimulates adipogenesis associated with activation of CB1 receptor and Akt signaling pathway in 3T3-L1 adipocyte. Biochem Biophys Res Commun 466:438–443. doi:10.1016/j.bbrc.2015.09.046

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by grants from the Baoen Wang liver fibrosis research fund, hepatitis prevention and treatment foundation of China (No.20110002), and Key discipline construction plan of Shanghai Public Health projects (No.12GWZX0903).

Authors contribution

Luoyan Ai and Xiaohan Wang conducted the experiments. Zhiwei Chen and Antao Xu analyzed data or performed statistical analysis. Qingqing Xu, Xiaoke Jiang, Dazhi Su, and Changwei Wu collected data and provided essential reagents. Qing Lin and Zhuping Fan designed the research. Luoyan Ai wrote the paper. Zhuping Fan had primary responsibility for final content. All authors have read and approved the final manuscript.

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    Authors and Affiliations

    1. Department of Health Manage Center, School of Medicine, Ren Ji Hospital, Shanghai Jiao Tong University, No. 160, Pujian Road, Shanghai, China

      Luoyan Ai, Xiaohan Wang, Zhiwei Chen, Qing Lin, Dazhi Su, Qingqing Xu, Changwei Wu, Xiaoke Jiang & Zhuping Fan

    2. Division of Gastroenterology and Hepatology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease; Key Laboratory of Gastroenterology & Hepatology, Ministry of Health, Shanghai Jiao Tong University, Shanghai, China

      Luoyan Ai, Dazhi Su, Qingqing Xu, Changwei Wu, Xiaoke Jiang & Antao Xu

    3. The First Hospital of Jiaxing, Jiaxing, Zhejiang, China

      Xiaohan Wang

    4. Division of Rheumatology, School of Medicine, Ren Ji Hospital, Shanghai Jiao Tong University, Shanghai, China

      Zhiwei Chen

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    1. Luoyan Ai
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    Luoyan Ai, Xiaohan Wang, and Zhiwei Chen contributed equally to this work.

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    Ai, L., Wang, X., Chen, Z. et al. A20 reduces lipid storage and inflammation in hypertrophic adipocytes via p38 and Akt signaling. Mol Cell Biochem 420, 73–83 (2016). https://doi.org/10.1007/s11010-016-2768-0

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