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Saturated fatty acid stimulates production of extracellular vesicles by renal tubular epithelial cells

  • Alyssa Cobbs
  • Xiaoming Chen
  • Yuanyuan Zhang
  • Jasmine George
  • Ming-bo Huang
  • Vincent Bond
  • Winston Thompson
  • Xueying ZhaoEmail author
Article

Abstract

Lipotoxicity, an accumulation of intracellular lipid metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of metabolic disease. Palmitic acid, a predominant lipid derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether palmitic acid would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated palmitic acid (PA), and BSA-conjugated oleic acid (OA) for 24–48 h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30–150 nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of lipid-induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of lipid-induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keywords

Extracellular vesicles Lipotoxicity Renal proximal tubules Palmitic acid 

Notes

Acknowledgements

This work was supported by the NIH SC1DK112151, NIH 5T32HL103104-07, NIH/NCRR/RCMI 8G12MD007602 and 8U54MD007588.

Compliance with ethical standards

Conflict of Interest

The authors declare no conflict of interest.

References

  1. 1.
    Saran R, Li Y, Robinson B, Abbott KC, Agodoa LY, Ayanian J, Bragg-Gresham J, Balkrishnan R, Chen JL, Cope E, Eggers PW, Gillen D, Gipson D, Hailpern SM, Hall YN, He K, Herman W, Heung M, Hirth RA, Hutton D, Jacobsen SJ, Kalantar-Zadeh K, Kovesdy CP, Lu Y, Molnar MZ, Morgenstern H, Nallamothu B, Nguyen DV, O’Hare AM, Plattner B, Pisoni R, Port FK, Rao P, Rhee CM, Sakhuja A, Schaubel DE, Selewski DT, Shahinian V, Sim JJ, Song P, Streja E, Kurella TM, Tentori F, White S, Woodside K, Hirth RA (2016) US Renal Data System 2015 Annual Data Report: Epidemiology of Kidney Disease in the United States, Am J Kidney Dis67: Svii, S1–Svii, 305.  https://doi.org/10.1053/j.ajkd.2015.12.014
  2. 2.
    Hager MR, Narla AD, Tannock LR (2017) Dyslipidemia in patients with chronic kidney disease. Rev Endocr Metab Disord 18:29–40.  https://doi.org/10.1007/s11154-016-9402-z CrossRefGoogle Scholar
  3. 3.
    Herman-Edelstein M, Scherzer P, Tobar A, Levi M, Gafter U (2014) Altered renal lipid metabolism and renal lipid accumulation in human diabetic nephropathy. J Lipid Res 55:561–572.  https://doi.org/10.1194/jlr.P040501 CrossRefGoogle Scholar
  4. 4.
    Jiang T, Wang XX, Scherzer P, Wilson P, Tallman J, Takahashi H, Li J, Iwahashi M, Sutherland E, Arend L, Levi M (2007) Farnesoid X receptor modulates renal lipid metabolism, fibrosis, and diabetic nephropathy. Diabetes 56:2485–2493.  https://doi.org/10.2337/db06-1642 CrossRefGoogle Scholar
  5. 5.
    Ormazabal V, Nair S, Elfeky O, Aguayo C, Salomon C, Zuniga FA (2018) Association between insulin resistance and the development of cardiovascular disease. Cardiovasc Diabetol 17:122.  https://doi.org/10.1186/s12933-018-0762-4 CrossRefGoogle Scholar
  6. 6.
    Prieur X, Roszer T, Ricote M (2010) Lipotoxicity in macrophages: evidence from diseases associated with the metabolic syndrome. Biochim Biophys Acta 1801:327–337.  https://doi.org/10.1016/j.bbalip.2009.09.017 CrossRefGoogle Scholar
  7. 7.
    Weinberg JM (2006) Lipotoxicity. Kidney Int 70:1560–1566.  https://doi.org/10.1038/sj.ki.5001834 CrossRefGoogle Scholar
  8. 8.
    Stadler K, Goldberg IJ, Susztak K (2015) The evolving understanding of the contribution of lipid metabolism to diabetic kidney disease. Curr Diabetes Rep 15:40.  https://doi.org/10.1007/s11892-015-0611-8 CrossRefGoogle Scholar
  9. 9.
    Hua W, Huang HZ, Tan LT, Wan JM, Gui HB, Zhao L, Ruan XZ, Chen XM, Du XG (2015) CD36 mediated fatty acid-induced podocyte apoptosis via oxidative stress. PLoS ONE 10:e0127507.  https://doi.org/10.1371/journal.pone.0127507 CrossRefGoogle Scholar
  10. 10.
    Lee E, Choi J, Lee HS (2017) Palmitate induces mitochondrial superoxide generation and activates AMPK in podocytes. J Cell Physiol 232:3209–3217.  https://doi.org/10.1002/jcp.25867 CrossRefGoogle Scholar
  11. 11.
    Sieber J, Lindenmeyer MT, Kampe K, Campbell KN, Cohen CD, Hopfer H, Mundel P, Jehle AW (2010) Regulation of podocyte survival and endoplasmic reticulum stress by fatty acids. Am J Physiol Renal Physiol 299:F821–F829.  https://doi.org/10.1152/ajprenal.00196.2010 CrossRefGoogle Scholar
  12. 12.
    Chen X, Li L, Liu X, Luo R, Liao G, Li L, Liu J, Cheng J, Lu Y, Chen Y (2018) Oleic acid protects saturated fatty acid mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis. Life Sci 203:291–304.  https://doi.org/10.1016/j.lfs.2018.04.022 CrossRefGoogle Scholar
  13. 13.
    Palomer X, Pizarro-Delgado J, Barroso E, Vazquez-Carrera M (2018) Palmitic and oleic acid: the Yin and yang of fatty acids in type 2 diabetes mellitus. Trends Endocrinol Metab 29:178–190.  https://doi.org/10.1016/j.tem.2017.11.009 CrossRefGoogle Scholar
  14. 14.
    Cobbs A, Ballou K, Chen X, George J, Zhao X (2018) Saturated fatty acids bound to albumin enhance osteopontin expression and cleavage in renal proximal tubular cells. Int J Physiol Pathophysiol Pharmacol 10:29–38Google Scholar
  15. 15.
    Xu R, Greening DW, Zhu HJ, Takahashi N, Simpson RJ (2016) Extracellular vesicle isolation and characterization: toward clinical application. J Clin Invest 126:1152–1162.  https://doi.org/10.1172/JCI81129 CrossRefGoogle Scholar
  16. 16.
    Hirsova P, Ibrahim SH, Krishnan A, Verma VK, Bronk SF, Werneburg NW, Charlton MR, Shah VH, Malhi H, Gores GJ (2016) Lipid-induced signaling causes release of inflammatory extracellular vesicles from hepatocytes. Gastroenterology 150:956–967.  https://doi.org/10.1053/j.gastro.2015.12.037 CrossRefGoogle Scholar
  17. 17.
    Lee YS, Kim SY, Ko E, Lee JH, Yi HS, Yoo YJ, Je J, Suh SJ, Jung YK, Kim JH, Seo YS, Yim HJ, Jeong WI, Yeon JE, Um SH, Byun KS (2017) Exosomes derived from palmitic acid-treated hepatocytes induce fibrotic activation of hepatic stellate cells. Sci Rep 7:3710.  https://doi.org/10.1038/s41598-017-03389-2 CrossRefGoogle Scholar
  18. 18.
    Lennon R, Pons D, Sabin MA, Wei C, Shield JP, Coward RJ, Tavare JM, Mathieson PW, Saleem MA, Welsh GI (2009) Saturated fatty acids induce insulin resistance in human podocytes: implications for diabetic nephropathy. Nephrol Dial Transplant 24:3288–3296.  https://doi.org/10.1093/ndt/gfp302 CrossRefGoogle Scholar
  19. 19.
    Sabin MA, Stewart CE, Crowne EC, Turner SJ, Hunt LP, Welsh GI, Grohmann MJ, Holly JM, Shield JP (2007) Fatty acid-induced defects in insulin signalling, in myotubes derived from children, are related to ceramide production from palmitate rather than the accumulation of intramyocellular lipid. J Cell Physiol 211:244–252.  https://doi.org/10.1002/jcp.20922 CrossRefGoogle Scholar
  20. 20.
    Mook S, Halkes CC, Bilecen S, Cabezas MC (2004) In vivo regulation of plasma free fatty acids in insulin resistance. Metabolism 53:1197–1201CrossRefGoogle Scholar
  21. 21.
    Murea M, Freedman BI, Parks JS, Antinozzi PA, Elbein SC, Ma L (2010) Lipotoxicity in diabetic nephropathy: the potential role of fatty acid oxidation. Clin J Am Soc Nephrol 5:2373–2379.  https://doi.org/10.2215/CJN.08160910 CrossRefGoogle Scholar
  22. 22.
    Nakamura K, Sawada K, Kinose Y, Yoshimura A, Toda A, Nakatsuka E, Hashimoto K, Mabuchi S, Morishige KI, Kurachi H, Lengyel E, Kimura T (2017) Exosomes promote ovarian cancer cell invasion through transfer of CD44 to peritoneal mesothelial cells. Mol Cancer Res 15:78–92.  https://doi.org/10.1158/1541-7786.MCR-16-0191 CrossRefGoogle Scholar
  23. 23.
    Huang MB, Gonzalez RR, Lillard J, Bond VC (2017) Secretion modification region-derived peptide blocks exosome release and mediates cell cycle arrest in breast cancer cells. Oncotarget 8:11302–11315.  https://doi.org/10.18632/oncotarget.14513 Google Scholar
  24. 24.
    Baumann JM, Kokabee L, Wang X, Sun Y, Wong J, Conklin DS (2015) Metabolic assays for detection of neutral fat stores. Bio Protoc 5:12CrossRefGoogle Scholar
  25. 25.
    Cohen BC, Shamay A, Argov-Argaman N (2015) Regulation of lipid droplet size in mammary epithelial cells by remodeling of membrane lipid composition-a potential mechanism. PLoS ONE 10:e0121645.  https://doi.org/10.1371/journal.pone.0121645 CrossRefGoogle Scholar
  26. 26.
    Akoumi A, Haffar T, Mousterji M, Kiss RS, Bousette N (2017) Palmitate mediated diacylglycerol accumulation causes endoplasmic reticulum stress, Plin2 degradation, and cell death in H9C2 cardiomyoblasts. Exp Cell Res 354:85–94.  https://doi.org/10.1016/j.yexcr.2017.03.032 CrossRefGoogle Scholar
  27. 27.
    Plotz T, Hartmann M, Lenzen S, Elsner M (2016) The role of lipid droplet formation in the protection of unsaturated fatty acids against palmitic acid induced lipotoxicity to rat insulin-producing cells. Nutr Metab (Lond) 13:16.  https://doi.org/10.1186/s12986-016-0076-z CrossRefGoogle Scholar
  28. 28.
    Borradaile NM, Han X, Harp JD, Gale SE, Ory DS, Schaffer JE (2006) Disruption of endoplasmic reticulum structure and integrity in lipotoxic cell death. J Lipid Res 47:2726–2737.  https://doi.org/10.1194/jlr.M600299-JLR200 CrossRefGoogle Scholar
  29. 29.
    Katsoulieris E, Mabley JG, Samai M, Sharpe MA, Green IC, Chatterjee PK (2010) Lipotoxicity in renal proximal tubular cells: relationship between endoplasmic reticulum stress and oxidative stress pathways. Free Radic Biol Med 48:1654–1662.  https://doi.org/10.1016/j.freeradbiomed.2010.03.021 CrossRefGoogle Scholar
  30. 30.
    Pomatto MAC, Gai C, Bussolati B, Camussi G (2017) Extracellular vesicles in renal pathophysiology. Front Mol Biosci 4:37.  https://doi.org/10.3389/fmolb.2017.00037 CrossRefGoogle Scholar
  31. 31.
    Ban LA, Shackel NA, McLennan SV (2016) Extracellular vesicles: a new frontier in biomarker discovery for non-alcoholic fatty liver disease. Int J Mol Sci 17:376.  https://doi.org/10.3390/ijms17030376 CrossRefGoogle Scholar
  32. 32.
    Harp D, Driss A, Mehrabi S, Chowdhury I, Xu W, Liu D, Garcia-Barrio M, Taylor RN, Gold B, Jefferson S, Sidell N, Thompson W (2016) Exosomes derived from endometriotic stromal cells have enhanced angiogenic effects in vitro. Cell Tissue Res 365:187–196.  https://doi.org/10.1007/s00441-016-2358-1 CrossRefGoogle Scholar
  33. 33.
    Yamamoto CM, Murakami T, Oakes ML, Mitsuhashi M, Kelly C, Henry RR, Sharma K (2018) Uromodulin mRNA from urinary extracellular vesicles correlate to kidney function decline in type 2 diabetes mellitus. Am J Nephrol 47:283–291.  https://doi.org/10.1159/000489129 CrossRefGoogle Scholar
  34. 34.
    Dominguez JM, Dominguez JH, Xie D, Kelly KJ (2018) Human extracellular microvesicles from renal tubules reverse kidney ischemia-reperfusion injury in rats. PLoS ONE 13:e0202550.  https://doi.org/10.1371/journal.pone.0202550 CrossRefGoogle Scholar
  35. 35.
    Gutierrez-Vazquez C, Villarroya-Beltri C, Mittelbrunn M, Sanchez-Madrid F (2013) Transfer of extracellular vesicles during immune cell-cell interactions. Immunol Rev 251:125–142.  https://doi.org/10.1111/imr.12013 CrossRefGoogle Scholar
  36. 36.
    De S, Kuwahara S, Hosojima M, Ishikawa T, Kaseda R, Sarkar P, Yoshioka Y, Kabasawa H, Iida T, Goto S, Toba K, Higuchi Y, Suzuki Y, Hara M, Kurosawa H, Narita I, Hirayama Y, Ochiya T, Saito A (2017) Exocytosis-mediated urinary full-length megalin excretion is linked with the pathogenesis of diabetic nephropathy. Diabetes 66:1391–1404.  https://doi.org/10.2337/db16-1031 CrossRefGoogle Scholar
  37. 37.
    Lv LL, Feng Y, Wen Y, Wu WJ, Ni HF, Li ZL, Zhou LT, Wang B, Zhang JD, Crowley SD, Liu BC (2018) Exosomal CCL2 from tubular epithelial cells is critical for albumin-induced tubulointerstitial inflammation. J Am Soc Nephrol 29:919–935.  https://doi.org/10.1681/ASN.2017050523 CrossRefGoogle Scholar
  38. 38.
    Musso G, Cassader M, Cohney S, De MF, Pinach S, Saba F, Gambino R (2016) Fatty liver and chronic kidney disease: novel mechanistic insights and therapeutic opportunities. Diabetes Care 39:1830–1845.  https://doi.org/10.2337/dc15-1182 CrossRefGoogle Scholar
  39. 39.
    Caruso S, Poon IKH (2018) Apoptotic cell-derived extracellular vesicles: more than just debris. Front Immunol 9:1486.  https://doi.org/10.3389/fimmu.2018.01486 CrossRefGoogle Scholar
  40. 40.
    Lynch C, Panagopoulou M, Gregory CD (2017) Extracellular vesicles arising from apoptotic cells in tumors: roles in cancer pathogenesis and potential clinical applications. Front Immunol 8:1174.  https://doi.org/10.3389/fimmu.2017.01174 CrossRefGoogle Scholar
  41. 41.
    Atkin-Smith GK, Poon IKH (2017) Disassembly of the dying: mechanisms and functions. Trends Cell Biol 27:151–162.  https://doi.org/10.1016/j.tcb.2016.08.011 CrossRefGoogle Scholar
  42. 42.
    Zhang W, Zhou X, Zhang H, Yao Q, Liu Y, Dong Z (2016) Extracellular vesicles in diagnosis and therapy of kidney diseases. Am J Physiol Renal Physiol 311:F844–F851.  https://doi.org/10.1152/ajprenal.00429.2016 CrossRefGoogle Scholar
  43. 43.
    Lu CC, Ma KL, Ruan XZ, Liu BC (2017) The emerging roles of microparticles in diabetic nephropathy. Int J Biol Sci 13:1118–1125.  https://doi.org/10.7150/ijbs.21140 CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of PhysiologyMorehouse School of MedicineAtlantaUSA
  2. 2.Department of Microbiology, Biochemistry & ImmunologyMorehouse School of MedicineAtlantaUSA

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