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Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo

자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과

  • Byun, Eui-Hong (Department of Food Science and Technology, Kongju National University)
  • 변의홍 (공주대학교 식품공학과)
  • Received : 2017.05.30
  • Accepted : 2017.07.21
  • Published : 2017.10.31

Abstract

This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.

본 연구는 자소엽 조다당 추출물(PCP)의 면역활성에 관하여 알아보기 위하여, 선천면역계의 대표적인 면역세포인 수지상세포 및 후천면역계의 대표적인 면역세포인 비장세포에 PCP를 처리하여 면역세포의 면역활성능에 관하여 측정하였다. 수지상세포에 PCP를 1, 5 및 $10{\mu}g/mL$의 농도를 처리한 결과, 세포생존율이 $103.4{\pm}3.8$, $108.8{\pm}2.1$, $117.8{\pm}3.3%$ (n =3)로 나타나 세포독성을 유발하지 않았으며, 주요 면역활성 인자인 산화질소의 분비능을 관찰한 결과, 각각 $2.7{\pm}0.2$, $4.5{\pm}0.2$, $7.3{\pm}0.3{\mu}M$ (n =3)로 농도 의존적으로 나타났다. 농도(1, 5 및 $10{\mu}g/mL$)별 PCP 처리구에서 사이토카인의 분비능을 관찰한 결과, TNF-${\alpha}$ ($372.3{\pm}0.32$, $604.8{\pm}0.54$$954.2{\pm}1.32pg/mL$), IL-6 ($508.4{\pm}0.39$, $761.5{\pm}1.34$$1038.5{\pm}1.67pg/mL$), IL-$1{\beta}$ ($314.5{\pm}1.04$, $524.8{\pm}1.89$$664.8{\pm}0.89pg/mL$), IL-12 ($321.4{\pm}0.94$, $832.5{\pm}0.85$$901.{\pm}0.94pg/mL$)가 유의적으로 증가되는 것으로 관찰되었다. 또한 후천면역에서 중요한 역할을 수행하는 면역 T 세포가 다량으로 분포하는 비장 조직으로부터 비장세포를 분리하여 PCP를 처리하였을 때, 세포 증식능이 유의적으로 증가하였으며, 면역활성을 유도하는 Th1 세포가 분비하는 사이토카인의 함량 또한 유의적으로 증가되는 것으로 나타났다. 이러한 결과로 미루어 볼 때 PCP의 처리는 선천면역뿐만 아니라 후천면역에 관여하는 다양한 면역세포의 활성화에 직간접적으로 관여하는 것으로 사료된다. 본 연구는 자소엽조다당 추출물의 면역활성 유도 효과에 관한 가능성을 제시하였고, 향후 자소엽 조다당 추출물을 분리 및 정제과정을 통하여 구조분석 및 정제된 조다당 추출물의 정확한 면역활성과 면역활성기전에 관한 면밀한 연구가 필요할 것으로 사료된다.

Keywords

References

  1. Hong HD, Cho CW, Rhee YK, Choi HD, Lee HS. Status of technology development using immune-modulating polysaccharide. Food Sci. Ind. 45: 1-11 (2012)
  2. Jiang MH, Zhu L, Jiang JG. Immunoregulatory actions of polysaccharides from Chinese herbal medicine. Expert Opin. Ther. Tar. 14: 1367-1402 (2010) https://doi.org/10.1517/14728222.2010.531010
  3. Cha JH, Lim EM. Effects of Gardeniae fructus on cytokines in mouse macrophage. J. Korean Obstet. Gynecol. 27: 1-16 (2014)
  4. Erwig LP, Rees AJ. Macrophage activation and programing and its role for macrophage function in glomerular inflammation. Kidney Blood Press. R. 22: 21-25 (1999) https://doi.org/10.1159/000025905
  5. But PPH, Kimura T, Guo JX, Sung CK, Han BH. International Collation of Traditional and Folk Medicine. World Scientific, Singapore. pp. 202-203 (1997)
  6. Ueda H, Yamazaki C, Yamazaki M. Inhibitory effect of perilla leaf extract and luteolin on mouse skin tumor promotion. Biol. Pharm. Bull. 26: 560-563 (2003) https://doi.org/10.1248/bpb.26.560
  7. Oh HA, Park C, Ahn HJ, Park YS, Kim HM. Effect of Perilla frutescens var. acuta Kudo and rosmarinic acid on allergic inflammatory reactions. Exp. Biol. Med. 236: 99-106 (2011) https://doi.org/10.1258/ebm.2010.010252
  8. Makino T, Furuta Y, Wakushima H, Fujii H, Saito K, Kano Y. Anti-allergic effect of Perilla frutescens and its active constituents. Phytother. Res. 17: 240-243 (2003) https://doi.org/10.1002/ptr.1115
  9. Choi UK, Lee OH, Lim SI, Kim YC. Optimization of antibacterial activity of Perilla frutescens var. acuta leaf against Pseudomonas aeruginosa using the evolutionary operation factorial design technique. Int. J. Mol. Sci. 11: 3922-3932 (2010) https://doi.org/10.3390/ijms11103922
  10. Meng L, Lozano YF, Gaydou EM, Li B. Antioxidant activities of polyphenols extracted from Perilla frutescens varieties. Molecules 14: 133-140 (2008) https://doi.org/10.3390/molecules14010133
  11. Kim DH, Kim YC, Choi UK. Optimization of antibacterial activity of Perilla frutescens var. acuta leaf against Staphylococcus aureus using evolutionary operation factorial design technique. Int. J. Mol. Sci. 12: 2395-2407 (2011) https://doi.org/10.3390/ijms12042395
  12. Fujita T, Funayoshi A, Nakayama M. A phenylpropanoid glucoside from Perilla frutescens. Phytochemistry 37: 543-546 (1994) https://doi.org/10.1016/0031-9422(94)85096-8
  13. Banno N, Akihisa T, Tokuda H, Yasukawa K, Higashihara H, Ukita M, Watanabe K, Kimura Y, Hasegawa J, Nishino H, Nishino H. Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effects. Biosci. Biotechnol. Biochem. 68: 85-90 (2004) https://doi.org/10.1271/bbb.68.85
  14. Zhao Q, Shepherd EG, Manson ME, Nelin LD, Sorokin A, Liu Y. The role of mitogen-activated protein kinase phosphatase-1 in the response of alveolar macrophages to lipopolysaccharide: Attenuation of proinflammatory cytokine biosynthesis via feedback control. J. Biol. Chem. 280: 8101-8108 (2005) https://doi.org/10.1074/jbc.M411760200
  15. Schepetkin IA, Quinn MT. Botanical polysaccharides: Macrophage immunomodulation and therapeutic potential. Int. Immunopharmacol. 6: 317-333 (2006) https://doi.org/10.1016/j.intimp.2005.10.005
  16. Sim SM, Im GH, Kim JW, Lee UY, Kim HW, Lee MU, Lee TS. The immuno-modulatory and antitumor effects of crude polysaccharides extracted from Daedaleopsis tricolor. Kor. J. Mycol. 31: 161-167 (2003) https://doi.org/10.4489/KJM.2003.31.3.161
  17. Lee HJ, Lee CW, Choi MS, Son DJ, Hong JT. Effects of esthetic essential oils on LPS-induced nitric oxide generation in murine macrophage RAW 264.7 cells. J. Soc. Cosmet. Sci. Korea 32: 111-116 (2006)
  18. Klimp AH, de Vries EG, Scherphof GL, Daemen T. A potential role of macrophage activation in the treatment of cancer. Crit. Rev. Oncol. Hematol. 44: 143-161 (2002) https://doi.org/10.1016/S1040-8428(01)00203-7
  19. Palmer RM, Ashton DS, Moncada S. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 333: 664-666 (1988) https://doi.org/10.1038/333664a0
  20. Chiou WF, Chou CJ, Chen CF. Camptothecin suppresses nitric oxide biosynthesis in RAW 264.7 macrophages. Life Sci. 69: 625-625 (2001) https://doi.org/10.1016/S0024-3205(01)01154-7
  21. Seo WG, Pae HO, Oh GS, Kim NY, Kwon TO, Shin MK, Chia KY, Chung HT. The aqueous extract of Rhodiola sachalinensis root enhances the expression of inducible nitric oxide synthase gene in RAW 264.7 macrophages. J. Ethnopharmacol. 76: 119-123 (2001) https://doi.org/10.1016/S0378-8741(01)00220-3
  22. Hibbs JB, Taintor RR, Vavrin I, Rachlin EM. Nitric oxide: A cytotoxic activated macrophage effector molecule. Biochem. Biophys. Res. Commun. 157: 87-82 (1998)
  23. Lee K, Sohn Y, Lee MJ, Cho HS, Jang MH, Han NY, Shin KW, Kim SH, Cho IH, Bu Y, Jung HS. Effects of Angelica acutiloba on mast cell-mediated allergic reactions in vitro and in vivo. Immunopharm. Immunot. 34: 571-577 (2012) https://doi.org/10.3109/08923973.2011.636048
  24. Calixto JB, Campos MM, Otuki MF, Santos AR. Anti-inflammatory compounds of plant origin. Part II. modulation of proinflammatory cytokines, chemokines and adhesion molecules. Planta Med. 70: 93-103 (2004) https://doi.org/10.1055/s-2004-815483
  25. Lee JH, Kim YS, Lim EM. Effects of Angelicae Pubescentis Radix water extract on immune property in RAW 264.7 macrophages. J. Korean Oriental Med. 32: 175-184 (2011)
  26. Byun EH. Comparison study of immunomodulatory activity of polysaccharide and ethanol extracted from Sargassum fulvellum. J. Korean Soc. Food Sci. Nutr. 44: 1621-1628 (2015) https://doi.org/10.3746/jkfn.2015.44.11.1621
  27. Kim HS, Kang JS. Preparation and characteristics of bread by medicinal herb composites with immunostimulating activity. J Korean Soc. Food Sci. Nutr. 37: 109-116 (2008) https://doi.org/10.3746/jkfn.2008.37.1.109
  28. Lee TS, Shim SM, Im KH, Kim JW, Lee UY, Shim MJ, Lee MW. Studies on immuno-modulatory and antitumor effects of crude polysaccharides extracted from Paecilomyces sinclairii. Kor. J. Mycol. 31: 155-160 (2003) https://doi.org/10.4489/KJM.2003.31.3.155
  29. Lee JK, Lee MK, Yun YP, Kim Y, Kim JS, Kim YS, Kim K, Han SS, Lee CK. Acemannan purified from Aloe vera induces phenotypic and functional maturation of immature dendritic cells. Int. Immunopharmacol. 1: 1275-1284 (2001) https://doi.org/10.1016/S1567-5769(01)00052-2
  30. Kikuchi T, Ohno N, Ohno T. Maturation of dendritic cells induced by Candida beta-D-glucan. Int. Immunopharmacol. 2: 1503-1508 (2002) https://doi.org/10.1016/S1567-5769(02)00084-X
  31. Piani A, Hossle JP, Birchler T, Siegrist CA, Heumann D, Davies G, Loeliger S, Seger R, Lauener RP. Expression of MHC class II molecules contributes to lipopolysaccharide responsiveness. Eur. J. Immunol. 30: 3140-3146 (2000) https://doi.org/10.1002/1521-4141(200011)30:11<3140::AID-IMMU3140>3.0.CO;2-O
  32. Hegde NR, Chevalier MS, Johnson DC. Viral inhibition of MHC class II antigen presentation. Trends Immunol. 24: 278-285 (2003) https://doi.org/10.1016/S1471-4906(03)00099-1
  33. Mo ZQ, Wang JL, Yang M, Ni LY, Wang HQ, Lao GF, Li YW, Li AX, Luo XC, Dan XM. Characterization and expression analysis of grouper (Epinephelus coioides) co-stimulatory molecules CD83 and CD80/86 postCryptocaryon irritans infection. Fish Shellfish Immun. 67: 467-474 (2017) https://doi.org/10.1016/j.fsi.2017.05.064
  34. Ryu HS, Kim J, Kim HS. Enhancing effect of Sorghum bicolor L. Moench (Sorghum, su-su) extracts on mouse spleen and macrophage cell activation. Korean J. Food Nutr. 19: 176-182 (2006)
  35. Shan BE, Yoshida Y, Kuroda E, Yamashita U. Immunomodulating activity of seaweed extract on human lymphocytes in vitro. Int. J. Immunopharmacol. 21: 59-70 (1999) https://doi.org/10.1016/S0192-0561(98)00063-0
  36. Medzhitov R. Toll-like receptors and innate immunity. Nat. Rev. Immunol. 1: 135-145 (2001) https://doi.org/10.1038/35100529
  37. Chen FY, Ye YP, Sun HX, Li HX, Shi H. Stemucronatoside L, a pregnane glycoside from the roots of Stephanotis mucronata, inhibits Th1/Th2 immune responses in vitro. Chem. Biodivers. 6: 916-923b (2009) https://doi.org/10.1002/cbdv.200800159
  38. Lee SH, Ahn WY. Chemical constituents of saccharides and tritperpenoids in the the korean native mistletoes. J. Korean Wood Sci. Technol. 24: 28-36 (1996)