Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Immune Enhancement Effects of Neutral Lipids, Glycolipids, Phospholipids from Halocynthia aurantium Tunic on RAW264.7 Macrophages

  • A-yeong Jang (Department of Wellness-Bio Industry, Gangneung-Wonju National University) ;
  • Weerawan Rod-in (Department of Marine Bio Food Science, Gangneung-Wonju National University) ;
  • Il-shik Shin (Department of Marine Bio Food Science, Gangneung-Wonju National University) ;
  • Woo Jung Park (Department of Wellness-Bio Industry, Gangneung-Wonju National University)
  • Received : 2023.07.05
  • Accepted : 2023.10.30
  • Published : 2024.02.28
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Abstract

Fractionated lipids of Halocynthia aurantium (Pyuridae) have been demonstrated to possess anti-inflammatory properties. However, their modulatory properties have not been reported yet. Thus, the objective of this study was to determine immune enhancing effects of fractionated lipids from H. aurantium tunic on macrophage cells. The tunic of H. aurantium was used to isolate total lipids, which were then subsequently separated into neutral lipids, glycolipids, and phospholipids. RAW264.7 cells were stimulated with different concentrations (0.5, 1.0, 2.0, and 4.0%) of each fractionated lipid. Cytotoxicity, production of NO, expression levels of immune-associated genes, and signaling pathways were then determined. Neutral lipids and glycolipids significantly stimulated NO and PGE2 production and expression levels of IL-1β, IL-6, TNF-α, and COX-2 in a dose-dependent manner, while phospholipids ineffectively induced NO production and mRNA expression. Furthermore, it was found that both neutral lipids and glycolipids increased NF-κB p-65, p38, ERK1/2, and JNK phosphorylation, suggesting that these lipids might enhance immunity by activating NF-κB and MAPK signaling pathways. In addition, H. aurantium lipids-induced TNF-α expression was decreased by blocking MAPK or NF-κB signaling pathways. Phagocytic activity of RAW 264.7 cells was also significantly enhanced by neutral lipids and glycolipids. These results suggest that neutral lipids and glycolipids from H. aurantium tunic have potential as immune-enhancing materials.

Keywords

Acknowledgement

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (RS-2023-00248832). This research project was also supported by the University Emphasis Research Institute Support Program (No.2018R1A61A03023584), which is funded by National Research Foundation of Korea. Additionally, this research were supported by the Korea Institute of Marine Science & Technology Promotion (KIMST) funded by the Ministry of Oceans and Fisheries (20220042, Korea Sea Grant Program: Gangwon Sea Grant) and supported by the Ministry of Small and Medium-sized Enterprises (SMEs) and Startups (MSS), Korea, under the "Regional Specialized lndustry Development Pius Prograrn (R&D, S3258709)" supervised by the Korea Technology and Information Promotion Agency for SMEs (TIPA).

References

  1. Kim YO, Park S, Nam BH, Jung YT, Kim DG, Bae KS, et al. 2014. Description of Lutimonas halocynthiae sp. nov., isolated from a golden sea squirt (Halocynthia aurantium), reclassification of Aestuariicola saemankumensis as Lutimonas saemankumensis comb. nov. and emended description of the genus Lutimonas. Int. J. Syst. Evol. Microbiol. 64: 1984-1990. https://doi.org/10.1099/ijs.0.059923-0
  2. Fomenko SE, Kushnerova NF, Lesnikova LN. 2013. Experimental assessment of the efficiency of erythrocyte membrane repair by an extract of the tunic of the ascidian purple sea squirt in carbon tetrachloride poisoning. Pharm. Chem. J. 46: 606-611. https://doi.org/10.1007/s11094-013-0855-z
  3. Lambert G, Karney RC, Rhee WY, Carman MR. 2016. Wild and cultured edible tunicates: a review. Manag. Biol. Invasions 7: 59-66. https://doi.org/10.3391/mbi.2016.7.1.08
  4. Hirose E, Ohtake SI, Azumi K. 2009. Morphological characterization of the tunic in the edible ascidian, Halocynthia roretzi (Drasche), with remarks on 'soft tunic syndrome' in aquaculture. J. Fish Dis. 32: 433-445. https://doi.org/10.1111/j.1365-2761.2009.01034.x
  5. Van Daele Y, Revol JF, Gaill F, Goffinet G. 1992. Characterization and supramolecular architecture of the cellulose-protein fibrils in the tunic of the sea peach (Halocynthia papillosa, Ascidiacea, Urochordata). Biol. Cell 76: 87-96. https://doi.org/10.1016/0248-4900(92)90198-A
  6. Song G, Delroisse J, Schoenaers D, Kim H, Nguyen TC, Horbelt N, et al. 2020. Structure and composition of the tunic in the sea pineapple Halocynthia roretzi: a complex cellulosic composite biomaterial. Acta Biomater. 111: 290-301. https://doi.org/10.1016/j.actbio.2020.04.038
  7. Arumugam V, Venkatesan M, Ramachandran S, Sundaresan U. 2018. Bioactive peptides from marine ascidians and future drug development-a review. J. Int. J. Peptide Res. Ther. 24: 13-18. https://doi.org/10.1007/s10989-017-9662-9
  8. Sawada T. 1992. Tunicates and their immune mechanism. Bull. Yamaguchi Med. Sch. 39: 83-88.
  9. Palanisamy SK, Rajendran NM, Marino A. 2017. Natural products diversity of marine ascidians (tunicates; ascidiacea) and successful drugs in clinical development. Nat. Prod. Bioprospect. 7: 1-111. https://doi.org/10.1007/s13659-016-0115-5
  10. Tabakaeva OV, Tabakaev AV. 2017. Amino acids and related compounds of the ascidian Halocynthia aurantium from the Sea of Japan. Chem. Nat. Compd. 53: 722-725. https://doi.org/10.1007/s10600-017-2099-8
  11. Lordan R, Tsoupras A, Zabetakis I. 2017. Phospholipids of animal and marine origin: structure, function, and anti-inflammatory properties. Molecules 22: 1964.
  12. Calder PC. 2015. Marine omega-3 fatty acids and inflammatory processes: effects, mechanisms and clinical relevance. Biochim. Biophys. Acta 1851: 469-484. https://doi.org/10.1016/j.bbalip.2014.08.010
  13. Xu B, Zhang XY, Jin HZ, Wang C. 2003. Determination of content of fat and composition of fatty acids in Ascidian. Chinese J. Mar. Drugs 22: 37-39.
  14. Ai-li J, Chang-hai W. 2006. Antioxidant properties of natural components from Salvia plebeia on oxidative stability of ascidian oil. Process Biochem. 41: 1111-1116. https://doi.org/10.1016/j.procbio.2005.12.001
  15. Viracaoundin I, Barnathan G, Gaydou EM, Aknin M. 2003. Phospholipid FA from indian ocean tunicates Eudistoma bituminis and Cystodytes violatinctus. Lipids 38: 85-88. https://doi.org/10.1007/s11745-003-1035-7
  16. Lee KH, Park CS, Hong BI, Jung WJ. 1993. Utilization of ascidian, Halocynthia roretzi-2. lipids of ascidian with seasonal and regional variation. Korean J. Fish Aquat. Sci. 26: 141-149.
  17. Mikami N, Hosokawa M, Miyashita K. 2010. Effects of sea squirt (Halocynthia roretzi) lipids on white adipose tissue weight and blood glucose in diabetic/obese KK-Ay  mice. Mol. Med. Rep. 3: 449-453. https://doi.org/10.3892/mmr_00000278
  18. Jang A, Monmai C, Rod-In W, Kim J-E, You S, Lee TH, et al. 2021. Immune-modulation effect of Halocynthia aurantium tunic lipid on RAW264.7 cells. Food Sci. Biotechnol. 31: 101-110. https://doi.org/10.1007/s10068-021-01017-4
  19. Lee IH, Lee YS, Kim CH, Kim CR, Hong T, Menzel L, et al. 2001. Dicynthaurin: an antimicrobial peptide from hemocytes of the solitary tunicate, Halocynthia aurantium. Biochim. Biophys. Acta 1527: 141-148. https://doi.org/10.1016/S0304-4165(01)00156-8
  20. Jang WS, Kim KN, Lee YS, Nam MH, Lee IH. 2002. Halocidin: a new antimicrobial peptide from hemocytes of the solitary tunicate, Halocynthia aurantium. FEBS Lett. 521: 81-86. https://doi.org/10.1016/S0014-5793(02)02827-2
  21. Jang WS, Kim CH, Kim KN, Park SY, Lee JH, Son SM, et al. 2003. Biological activities of synthetic analogs of halocidin, an antimicrobial peptide from the tunicate Halocynthia aurantium. Antimicrob. Agents Chemother. 47: 2481-2486. https://doi.org/10.1128/AAC.47.8.2481-2486.2003
  22. Jo JE, Kim KH, Yoon MH, Kim NY, Lee C, Yook HS. 2010. Quality characteristics and antioxidant activity research of Halocynthia roretzi and Halocynthia aurantium. J. Korean Soc. Food Sci. Nutr. 39: 1481-1486. https://doi.org/10.3746/jkfn.2010.39.10.1481
  23. Chiji H, Hayashi C, Matsumoto M. 2001. Gastroprotective effect of Ascidian, Halocynthia aurantium (Akaboya), extract on acute gastric hemorrhagic lesions in rats, pp. 463-466. In Sawada H, Yokosawa H, Lambert CC (eds.), The Biology of Ascidians, Ed. Springer, Tokyo
  24. Monmai C, Go SH, Shin IS, You SG, Lee H, Kang SB, et al. 2018. Immune-enhancement and anti-inflammatory activities of fatty acids extracted from Halocynthia aurantium tunic in RAW264.7 cells. Mar. Drugs 16: 309.
  25. Han L, Yu J, Chen Y, Cheng D, Wang X, Wang C. 2018. Immunomodulatory activity of docosahexenoic acid on RAW264.7 cells activation through GPR120-mediated signaling pathway. J. Agric. Food Chem. 66: 926-934. https://doi.org/10.1021/acs.jafc.7b05894
  26. Lim JH, Choi GS, Monmai C, Rod-in W, Jang Ay, Park WJ. 2021. Immunomodulatory activities of Ammodytes personatus egg lipid in RAW264.7 cells. Molecules 26: 6027.
  27. Jang A, Rod-in W, Monmai C, Choi GS, Park WJ. 2022. Anti-inflammatory effects of neutral lipids, glycolipids, phospholipids from Halocynthia aurantium tunic by suppressing the activation of NF-κB and MAPKs in LPS-stimulated RAW264.7 macrophages. PLoS One 17: e0270794.
  28. Bligh EG, Dyer WJ. 1959. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37: 911-917. https://doi.org/10.1139/y59-099
  29. Christie W. 1982. Lipid analysis, pp. 2nd Ed. Pergamon Press, New York.
  30. Kim YS, Kim EK, Nawarathna WP, Dong X, Shin WB, Park JS, et al. 2017. Immune-stimulatory effects of Althaea rosea flower extracts through the MAPK signaling pathway in RAW264.7 cells. Molecules 22: 679.
  31. Xie Y, Wang L, Sun H, Wang Y, Yang Z, Zhang G, et al. 2019. Polysaccharide from alfalfa activates RAW 264.7 macrophages through MAPK and NF-κB signaling pathways. Int. J. Biol. Macromol. 126: 960-968. https://doi.org/10.1016/j.ijbiomac.2018.12.227
  32. Deng JJ, Li ZQ, Mo ZQ, Xu S, Mao HH, Shi D, et al. 2020. Immunomodulatory effects of N-acetyl chitooligosaccharides on RAW264.7 macrophages. Mar. Drugs 18: 421.
  33. Lee SH, Lee YP, Kim SY, Jeong MS, Lee MJ, Kang HW, et al. 2008. Inhibition of LPS-induced cyclooxygenase 2 and nitric oxide production by transduced PEP-1-PTEN fusion protein in RAW264.7 macrophage cells. Exp. Mol. Med. 40: 629-638. https://doi.org/10.3858/emm.2008.40.6.629
  34. Kim S, Lee CH, Yeo J-Y, Hwang KW, Park S-Y. 2022. Immunostimulatory activity of stem bark of Kalopanax pictus in RAW 264.7 macrophage. J. Herb. Med. 32: 100504.
  35. He C, Lin HY, Wang CC, Zhang M, Lin YY, Huang FY, et al. 2019. Exopolysaccharide from Paecilomyces lilacinus modulates macrophage activities through the TLR4/NF-κB/MAPK pathway. Mol. Med. Rep. 20: 4943-4952. https://doi.org/10.3892/mmr.2019.10746
  36. Eo HJ, Shin H, Song JH, Park GH. 2021. Immuno-enhancing effects of fruit of Actinidia polygama in macrophages. Food Agric. Immunol. 32: 754-765. https://doi.org/10.1080/09540105.2021.1982868
  37. Son HJ, Eo HJ, Park GH, Jeong JB. 2021. Heracleum moellendorffii root extracts exert immunostimulatory activity through TLR2/4-dependent MAPK activation in mouse macrophages, RAW264.7 cells. Food Sci. Nutr. 9: 514-521. https://doi.org/10.1002/fsn3.2020