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The immune enhancement effect of Nelumbo nucifera Gaertner Seed Extract (NSE) in murine macrophage RAW 264.7 cells

RAW 264.7 대식세포에서 연자육 추출물(Nelumbo nucifera Gaertner Seed Extract, NSE)의 면역 증강 효과

  • Se Jeong Kim (School of Food Science and Biotechnology, Kyungpook National University) ;
  • San Kim (School of Food Science and Biotechnology, Kyungpook National University) ;
  • Se Hyeon Jang (School of Food Science and Biotechnology, Kyungpook National University) ;
  • Sung Ran Yoon (Division of Agricultural Environment Research, Gyeongsangbuk-do Agricultural Research & Extension services) ;
  • Bo Ram So (Division of Agricultural Environment Research, Gyeongsangbuk-do Agricultural Research & Extension services) ;
  • Jeong Min Park (Division of Agricultural Environment Research, Gyeongsangbuk-do Agricultural Research & Extension services) ;
  • Jung A Ryu (ivision of Agricultural Environment Research, Gyeongsangbuk-do Agricultural Research & Extension services) ;
  • Sung Keun Jung (School of Food Science and Biotechnology, Kyungpook National University)
  • Received : 2022.12.06
  • Accepted : 2023.01.19
  • Published : 2023.12.31

Abstract

Since the global shock caused by COVID-19, interest in immune-enhancing materials is rapidly increasing, therefore, the development of novel materials is necessary from the industrial and health perspectives. In this study, we selected Nelumbo nucifera Gaertner Seed Extract (NSE) and evaluated immune enhancement effect by using RAW 264.7 murine macrophage cells. NSE significantly up-regulated production of nitric oxide and reactive oxygen species without affecting cell viability in RAW 264.7 cells. Additionally, NSE exhibited an increase of inducible nitric oxide synthase and cyclooxygenase-2 expression in RAW 264.7 cells. The enzyme-linked immunosorbent assay results showed that NSE-treatment significantly enhanced production of interleukin 6 and tumor necrosis factor-α in RAW 264.7 cells. Furthermore, we observed that NSE significantly up-regulated phosphorylation of p65, I kappa B kinase α/β, and I kappa B (IκB) α as well as down-regulation of IκB α expression in RAW 264.7 cells. Our findings indicate that NSE could be the potential health-functional food material with capacity of improving immunity via Nuclear factor-kappa B signaling pathway.

코로나바이러스-19로 인한 세계적인 펜데믹 이후 면역력 강화소재에 대한 관심이 급격히 증가하고 있으므로 산업적, 건강적 측면에서 새로운 소재의 개발이 필요하다. 본 연구에서는 연자육 추출물을 소재로 선정하고 RAW 264.7 쥐 대식세포를 이용하여 면역증진 효과를 평가하였다. 연자육 추출물은 RAW 264.7 세포에서 세포 생존력에 독성을 나타내지 않으면서 nitric oxide 및 reactive oxygen species의 생산을 상향 조절하였다. 또한 연자육 추출물은 RAW 264.7 세포에서 inducible nitric oxide synthase 및 cyclooxygenase-2 발현을 크게 증가시켰다. Enzyme-linked immunosorbent assay 결과에서는 연자육 추출물의 처리가 RAW 264.7 세포에서 interleukin 6 및 tumor necrosis factor-α의 생성을 유의미하게 향상시키는 것으로 나타났다. 또한 연자육 추출물이 p65, I kappa B kinase α/β, 및 I kappa B (IκB) α의 인산화를 크게 상향 조절하고, RAW 264.7 세포에서 IκB α의 발현을 하향 조절하였다. 우리의 연구 결과는 연자육 추출물이 Nuclear factor-kappa B 신호전달 경로를 통해 면역력을 향상시킬 수 있는 잠재적인 건강기능식품 소재가 될 수 있음을 나타낸다.

Keywords

Acknowledgement

본 연구는 2022년 경상북도기술원의 1팀 1교수 책임제 공동연구 '경북식재료활용K-면역농식품기술개발' (과제번호: LP0048882022) 연구비 지원에 의하여 수행되었습니다.

References

  1. Cassandra C, Elizabeth K, Leonard HC (2022) Long COVID and rheumatology: Clinical, diagnostic, and therapeutic implications. Best Pract Res Clin Rheumatol 101794. doi: 10.1016/j.berh.2022.101794
  2. Lee J, Kwan Y, Lee JY, Shin JI, Lee K H Lee, Hong SH, Han YJ, Kronbichler A, Smith L, Koyanagi A, Jacob L, Choi S, Ghayda RA, Park M (2021) Public Interest in Immunity and the Justification for Intervention in the Early Stages of the COVID-19 Pandemic: Analysis of Google Trends Data. J Med Internet Res 23(6): e26368. doi: 10.2196/26368
  3. Tripathy S, Verma DK, Thakur M, Patel AR, Srivastav PP, Singh S, Gupta AK, Chavez-Gonzalez ML, Aguilar CN, Chakravorty N, Verma HK, Utama GL (2021) Curcumin Extraction, Isolation, Quantification and Its Application in Functional Foods: A Review With a Focus on Immune Enhancement Activities and COVID-19. Front Nutr 21(8): 747956. doi: 10.3389/fnut.2021.747956
  4. Dong Y, Jiang W, Wu P, Liu Y, Kuang S, Tang L, Tang W, Zhou X, Feng L (2022) Novel Insight Into Nutritional Regulation in Enhancement of Immune Status and Mediation of Inflammation Dynamics Integrated Study In Vivo and In Vitro of Teleost Grass Carp (Ctenopharyngodon idella): Administration of Threonine. Front Immunol 13: 770969. doi: 10.3389/fimmu.2022.770969
  5. Huang Q, Wang J, Zong R, Wu D, Jin C (2022) A Water-Soluble Polysaccharide from the Fibrous Root of Anemarrhena asphodeloides Bge. and Its Immune Enhancement Effect in Vivo and in Vitro. Evid Based Complement Alternat Med 2022: 8723119. doi: 10.1155/2022/8723119
  6. Peng X, Feng L, Jiang W, Wu P, Liu Y, Jiang J, Kuang S, Tang L, Zhou X (2019) Supplementation exogenous bile acid improved growth and intestinal immune function associated with NF-kappaB and TOR signalling pathways in on-growing grass carp (Ctenopharyngodon idella): Enhancement the effect of protein-sparing by dietary lipid. Fish Shellfish Immunol 92: 552-569. doi: 10.1016/j.fsi.2019.06.047
  7. Li C, Dong Z, Zhang B, Huanga Q, Liu G, Fu X (2020) Structural characterization and immune enhancement activity of a novel polysaccharide from Moringa oleifera leaves. Carbohydr Polym 234: 115897. doi: 10.1016/j.carbpol.2020.115897
  8. Ding H, Shi J, Wang Y, Guo J, Zhao J, Dong L (2011) Neferine inhibits cultured hepatic stellate cell activation and facilitates apoptosis: A possible molecular mechanism. Eur J Pharmacol 650(1): 163-169. doi: 10.1016/j.ejphar.2010.10.025
  9. Sugimoto Y, Nishimura K, Itoh A, Tanahashi T, Nakajima H, Oshiro H, Sun D, Toda T, Yamada J (2015) Serotonergic mechanisms are involved in antidepressant-like effects of bisbenzylisoquinolines liensinine and its analogs isolated from the embryo of Nelumbo nucifera Gaertner seeds in mice. J Pharm Pharmacol 67(12): 1716-1722. doi: 10.1111/jphp.12473
  10. Youn UJ, Lee J, Lee Y J, Nam J W, Bae H, Seo E (2010) Regulation of the 5-HT3A receptor-mediated current by alkyl 4-hydroxybenzoates isolated from the seeds of Nelumbo nucifera. Chem Biodivers 7(9): 2296-2302. doi: 10.1002/cbdv.200900393
  11. Bharathi Priya L, Huang C, Hu R, Balasubramanian B, Baskaran R (2021) An updated review on pharmacological properties of neferine-A bisbenzylisoquinoline alkaloid from Nelumbo nucifera. J Food Biochem 45(12): e13986. doi: 10.1111/jfbc.13986
  12. So BR, Bach TT, Paik JH, Jung SK (2020) Kmeria duperreana (Pierre) Dandy Extract Suppresses LPS-Induced iNOS and NO via Regulation of NF-κB Pathways and p38 in Murin Macrophage RAW 264.7 Cells. Prev Nutr Food Sci 25(2): 166-172. doi: 10.3746/pnf.2020.25.2.166
  13. Kim MJ, Kim DW, Kim JG, Shin Y, Jung SK, Kim Y-J (2021) Analysis of the Chemical, Antioxidant, and Anti-Inflammatory Properties of Pink Pepper (Schinus molle L.). Antioxidants 10(7): 1062. doi: 10.3390/antiox10071062
  14. Lee WJ, Li WY, Lee SW, Jung SK (2021) Anti-Inflammatory and Antioxidant Effects of Soroseris hirsuta Extract by Regulating iNOS/NF-κB and NRF2/HO-1 Pathways in Murine Macrophage RAW 264.7 Cells. Applied Sciences 11(10): 4711. doi: 10.3390/app11104711
  15. Yeung YK, Kang Y, So BR, Jung SK, Chang YH (2021) Structural, antioxidant, prebiotic and anti-inflammatory properties of pectic oligosaccharides hydrolyzed from okra pectin by Fenton reaction. Food Hydrocolloids 118: 106779. doi: 10.1016/j.foodhyd.2021.106779
  16. Song JH, Kwak S, Kim H, Jun W, Lee J, Yoon HG, Kim Y, Choi KC (2019) Dendropanax morbifera Branch Water Extract Increases the Immunostimulatory Activity of RAW264.7 Macrophages and Primary Mouse Splenocytes. J Med Food 22(11): 1136-1145. doi: 10.1089/jmf.2019.4424
  17. Trinh TA, Park J , Oh JH , Park JS, Lee D, Kim CE, Choi HS, Kim SB, Hwang GS, Koo BA, Kang KS (2020) Effect of Herbal Formulation on Immune Response Enhancement in RAW 264.7 Macrophages. Biomolecules 10(3): 424. doi: 10.3390/biom10030424
  18. Garcia-Ortiz A, Serrador JM (2018) Nitric Oxide Signaling in T Cell-Mediated Immunity. Trends Mol Med 24(4): 412-427. doi: 10.3390/biom10030424
  19. Somensia N, Rabelo TK, Guimaraes AG, Quintans-Junior LJ, Souza Araujo AA, Moreira JCF, Gelain DP (2019) Carvacrol suppresses LPS-induced pro-inflammatory activation in RAW 264.7 macrophages through ERK1/2 and NF-kB pathway. Int Immunopharmacol 75: 105743. doi: 10.1016/j.intimp.2019.105743
  20. Facchin BM, Reis GO, Vieira GN, Bramorski Mohr ET, Salvan da Rosa J, Kretzer IF, Demarchi IG, Dalmarco EM (2022) Inflammatory biomarkers on an LPS-induced RAW 264.7 cell model: a systematic review and meta-analysis. Inflamm Res 71(7-8): 741-758. doi: 10.1007/s00011-022-01584-0
  21. Hong YH, Yi YS, Han SY, Aziz N, Kim HG, Park SH, Hossain MA, Baik KS, Choi SY, Lee J, Kim JH, Cho JY (2019) Morinda citrifolia noni water extract enhances innate and adaptive immune responses in healthy mice, ex vivo, and in vitro. Phytother Res 33(3): 676-689. doi: 10.1002/ptr.6256
  22. Lee SM, Choi W, Shi WR, Kim YH, Min J (2021) Enhanced Immune Response by Vacuoles isolated from Saccharomyces cerevisiae in RAW 264.7 Macrophages. Biosci Rep 41(9): BSR20211158. doi: 10.1042/BSR20211158
  23. Hong SH, Ku JM, Kim HI, Ahn CW, Park SH, Seo HS, Shin YC, Ko SG (2017) The immune-enhancing activity of Cervus nippon mantchuricus extract (NGE) in RAW264.7 macrophage cells and immunosuppressed mice. Food Res Int, 99(Pt 1): 623-629. doi: 10.1016/j.foodres.2017.06.053
  24. Ogasawara N, Matsushima M, Kawamura N, Atsumi K. Yamaguchi T, Ochi H, Kusatsugu Y, Oyabu S, Hashimoto N, Hasegawa Y, Ueyama J, Kawabe T (2017) Modulation of immunological activity on macrophages induced by diazinon. Toxicology 379: 22-30. doi: 10.1016/j.tox.2017.01.014
  25. Monmai C, Go SH, Shin I, You SG, Lee H, Kang SB, Park WJ (2018) Immune Enhancement Effect of Asterias amurensis Fatty Acids through NF-kappaB and MAPK Pathways on RAW 264.7 Cells. J Microbiol Biotechnol 28(3): 349-356. doi: 10.4014/jmb.1709.09005
  26. Monmai C, Go SH, Shin IS, You SG, Lee H, Kang SB, Park WJ (2018) Immune-Enhancement and Anti-Inflammatory Activities of Fatty Acids Extracted from Halocynthia aurantium Tunic in RAW264.7 Cells. Mar Drugs 16(9): 309. doi: 10.3390/md16090309
  27. Hughes JE, Srinivasan S, Lynch KR, Proia RL, Ferdek P, Hedrick CC (2008) Sphingosine-1-phosphate induces an antiinflammatory phenotype in macrophages. Circ Res 102(8): 950-958. doi: 10.1161/CIRCRESAHA.107.170779
  28. Monmai C, Kim JS, Baek SH (2022) Transgenic Rice Seed Extracts Exert Immunomodulatory Effects by Modulating Immune-Related Biomarkers in RAW264.7 Macrophage Cells. Nutrients 14(19): 4143. doi: 10.3390/nu14194143