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Heat-Treated Paraprobiotic Latilactobacillus sakei KU15041 and Latilactobacillus curvatus KU15003 Show an Antioxidant and Immunostimulatory Effect

  • Jun-Hyun Hyun (Department of Food Science and Biotechnology of Animal Resources, Konkuk University) ;
  • Im-Kyung Woo (Department of Food Science and Biotechnology of Animal Resources, Konkuk University) ;
  • Kee-Tae Kim (Research Institute, WithBio Inc.) ;
  • Young-Seo Park (Department of Food Science and Biotechnology, Gachon University) ;
  • Dae-Kyung Kang (Department of Animal Biotechnology, Dankook University) ;
  • Na-Kyoung Lee (Department of Food Science and Biotechnology of Animal Resources, Konkuk University) ;
  • Hyun-Dong Paik (Department of Food Science and Biotechnology of Animal Resources, Konkuk University)
  • Received : 2023.09.05
  • Accepted : 2023.09.25
  • Published : 2024.02.28

Abstract

The lactic acid bacteria, including Latilactobacillus sakei and Latilactobacillus curvatus, have been widely studied for their preventive and therapeutic effects. In this study, the underlying mechanism of action for the antioxidant and immunostimulatory effects of two strains of heat-treated paraprobiotics was examined. Heat-treated L. sakei KU15041 and L. curvatus KU15003 showed higher radical scavenging activity in both the 2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assays than the commercial probiotic strain LGG. In addition, treatment with these two strains exhibited immunostimulatory effects in RAW 264.7 macrophages, with L. curvatus KU15003 showing a slightly higher effect. Additionally, they promoted phagocytosis and NO production in RAW 264.7 cells without any cytotoxicity. Moreover, the expression of tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 was upregulated. These strains resulted in an increased expression of inducible nitric oxide synthase and cyclooxygenase-2. Moreover, the nuclear factor-κB and mitogen-activated protein kinase signaling pathways were stimulated by these strains. These findings suggest the potential of using L. sakei KU15041 and L. curvatus KU15003 in food or by themselves as probiotics with antioxidant and immune-enhancing properties.

Keywords

Acknowledgement

This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, and Forestry (IPET) through the High Value-added Food Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (#321035-5).

References

  1. Singh S, Singh D. 2023. Current and future prospects of flavonoids for human immune system. In: Jesharwani RK, Keservani RK, Keservani RK, Sharma AK (eds.) Nutraceuticals and functional foods in immunomodulators. Springer, Singapore. 
  2. Iddir M, Brito A, Dingeo G, Fernandez Del Campo SS, Samouda H, La Frano MR, et al. 2020. Strengthening the immune system and reducing inflammation and oxidative stress through diet and nutrition: considerations during the COVID-19 crisis. Nutrients 12: 1562. 
  3. Aliko V, Qirjo M, Sula E, Morina V, Faggio C. 2018. Antioxidant defense system, immune response and erythron profile modulation in gold fish, Carassius auratus, after acute manganese treatment. Fish Shellfish Immunol. 76: 101-109.  https://doi.org/10.1016/j.fsi.2018.02.042
  4. Locati M, Mantovani A, Sica A. 2013. Macrophage activation and polarization as an adaptive component of innate immunity. Adv. Immunol. 120: 163-184.  https://doi.org/10.1016/B978-0-12-417028-5.00006-5
  5. Lee JH, Phelan P, Shin M, Oh BC, Han X, Im SS, et al. 2018. SREBP-1a-stimulated lipid synthesis is required for macrophage phagocytosis downstream of TLR4-directed mTORC1. Proc. Natl. Acad. Sci. USA 115: E12228-E12234.  https://doi.org/10.1073/pnas.1813458115
  6. Moon PD, Lee JS, Kim HY, Han NR, Kang I, Kim HM, et al. 2019. Heat-treated Lactobacillus plantarum increases the immune responses through activation of natural killer cells and macrophages on in vivo and in vitro models. J. Med. Microbiol. 68: 467-474.  https://doi.org/10.1099/jmm.0.000938
  7. Liu Y, Li QZ, Zhou XW. 2021. Immunostimulatory effects of the intracellular polysaccharides isolated from liquid culture of Ophiocordyceps sinensis (Ascomycetes) on RAW264. 7 cells via the MAPK and PI3K/Akt signaling pathways. J. Ethnopharmacol. 275: 114130. 
  8. Kim WJ, Yu HS, Lee NK, Paik HD. 2022. Levilactobacillus brevis KU15151 inhibits Staphylococcus aureus lipoteichoic acid-induced inflammation in RAW 264.7 macrophages. Probiotics Antimicrob. Proteins 14: 767-777.  https://doi.org/10.1007/s12602-022-09949-x
  9. Jeon HL, Lee NK, Yang SJ, Kim WS, Paik HD. 2017. Probiotic characterization of Bacillus subtilis P223 isolated from kimchi. Food Sci. Biotechnol. 26: 1641-1648.  https://doi.org/10.1007/s10068-017-0148-5
  10. Zagorec M, Champomier-Verges MC. 2017. Lactobacillus sakei: a starter for sausage fermentation, a protective culture for meat products. Microorganisms 5: 56. 
  11. Lorenzo JM, Fontan MCG, Cachaldora A, Franco I, Carballo J. 2010. Study of the lactic acid bacteria throughout the manufacture of dry-cured lacon (a Spanish traditional meat product). Effect of some additives. Food Microbiol. 27: 229-235.  https://doi.org/10.1016/j.fm.2009.10.003
  12. Tang G, Zhang L. 2022. Update on strategies of probiotics for the prevention and treatment of colorectal cancer. Nutr. Cancer 74: 27-38.  https://doi.org/10.1080/01635581.2020.1865420
  13. Lee NK, Paik HD. 2021. Prophylactic effects of probiotics on respiratory viruses including COVID-19: a review. Food Sci. Biotechnol. 30: 773-781.  https://doi.org/10.1007/s10068-021-00913-z
  14. Lee NK, Park YS, Kang DK, Paik HD. 2023. Paraprobiotics: definition, manufacturing methods, and functionality. Food Sci. Biotechnol. 32: 1981-1991. 
  15. Kim JG, Dong X, Park SH, Bayazid AB, Jeoung SA, Lim BO. 2021. Bioconversion of black rice and blueberry regulate immunity system through regulation of MAPKs, NF-kB in RAW264.7 macrophage cells. Food Agric. Immunol. 32: 471-481.  https://doi.org/10.1080/09540105.2021.1956434
  16. Song MW, Park JY, Lee HS, Kim KT, Paik HD. 2021. Co-fermentation by Lactobacillus brevis B7 improves the antioxidant and immunomodulatory activities of hydroponic ginseng-fortified yogurt. Antioxidants 10: 1447. 
  17. Choi GH, Bock HJ, Lee NK, Paik HD. 2022. Soy yogurt using Lactobacillus plantarum 200655 and fructooligosaccharides: neuroprotective effects against oxidative stress. J. Food Sci. Technol. 59: 4870-4879.  https://doi.org/10.1007/s13197-022-05575-1
  18. Hyun JH, Yu HS, Woo IK, Lee GW, Lee NK, Paik HD. 2023. Anti-inflammatory activities of Levilactobacillus brevis KU15147 in RAW 264.7 cells stimulated with lipopolysaccharide on attenuating NF-κB, AP-1, and MAPK signaling pathways. Food Sci. Biotechnol. https://doi.org/10.1007/s10068-023-01318-w 
  19. Lee J, Kim HJ, Nguyen TTH, Kim SC, Ree J, Choi TG, Sohng JK, Park YI. 2020. Emodin 8-O-glucoside primes macrophages more strongly than emodin aglycone via activation of phagocytic activity and TLR-2/MAPK/NF-κB signalling pathway. Int. Immunopharmacol. 88: 106936. 
  20. Yu HS, Lee NK, Choi AJ, Choe JS, Bae CH, Paik HD. 2019 Anti-inflammatory potential of probiotic strain Weissella cibaria JW15 isolated from kimchi through regulation of NF-κB and MAPKs pathways in LPS-induced RAW 264.7 cells. J. Microbiol. Biotechnol. 29: 1022-1032.  https://doi.org/10.4014/jmb.1903.03014
  21. Ohteki T, Suzue K, Maki C, Ota T, Koyasu S. 2001. Critical role of IL-15-IL-15R for antigen-presenting cell functions in the innate immune response. Nat. Immunol. 2: 1138-1143.  https://doi.org/10.1038/ni729
  22. Pluddemann A, Mukhopadhyay S, Gordon S. 2011. Innate immunity to intracellular pathogens: macrophage receptors and responses to microbial entry. Immunol. Rev. 240: 11-24.  https://doi.org/10.1111/j.1600-065X.2010.00989.x
  23. Cunningham-Rundles S, McNeeley DF, Moon A. 2005. Mechanisms of nutrient modulation of the immune response. J. Allergy Clin. Immunol. 115: 1119-1128.  https://doi.org/10.1016/j.jaci.2005.04.036
  24. Medzhitov R. 2007. Recognition of microorganisms and activation of the immune response. Nature 449: 819-826.  https://doi.org/10.1038/nature06246
  25. Malhotra N, Kang J. 2013. SMAD regulatory networks construct a balanced immune system. Immunology 139: 1-10.  https://doi.org/10.1111/imm.12076
  26. Takahashi LS, Biller-Takahashi JD, Mansano CFM, Urbinati EC, Gimbo RY, Saita MV. 2017. Long-term organic selenium supplementation overcomes the trade-off between immune and antioxidant systems in pacu (Piaractus mesopotamicus). Fish Shellfish Immunol. 60: 311-317.  https://doi.org/10.1016/j.fsi.2016.11.060
  27. Cristofori F, Dargenio VN, Dargenio C, Miniello VL, Barone M, Francavilla R. 2021. Anti-inflammatory and immunomodulatory effects of probiotics in gut inflammation: a door to the body. Front. Immunol. 12: 578386. 
  28. Yousefi B, Eslami M, Ghasemian A, Kokhaei P, Salek Farrokhi A, Darabi N. 2019. Probiotics importance and their immunomodulatory properties. J. Cell. Physiol. 234: 8008-8018.  https://doi.org/10.1002/jcp.27559
  29. Plaza-Diaz J, Ruiz-Ojeda FJ, Gil-Campos M, Gil A. 2019. Mechanisms of action of probiotics. Adv. Nutr. 10: S49-S66.  https://doi.org/10.1093/advances/nmy063
  30. Kang CH, Kim JS, Kim H, Park HM, Paek NS. 2021. Heat-killed lactic acid bacteria inhibit nitric oxide production via inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells. Probiotics Antimicrob. Proteins 13: 1530-1538.  https://doi.org/10.1007/s12602-021-09781-9
  31. Pique N, Berlanga M, Minana-Galbis D. 2019. Health benefits of heat-killed (Tyndallized) probiotics: an overview. Int. J. Mol. Sci. 20: 2534. 
  32. Song MW, Chung Y, Kim KT, Hong WS, Chang HJ, Paik HD. 2020. Probiotic characteristics of Lactobacillus brevis B13-2 isolated from kimchi and investigation of antioxidant and immune-modulating abilities of its heat-killed cells. LWT-Food Sci. Technol. 128: 109452. 
  33. Ivan AL, Campanini MZ, Martinez RM, Ferreira VS, Steffen VS, Vicentini FT, Vilela FM, Martins FS, Zarpelon AC, Cunha TM. 2014. Pyrrolidine dithiocarbamate inhibits UVB-induced skin inflammation and oxidative stress in hairless mice and exhibits antioxidant activity in vitro. J. Photochem. Photobiol. B Biol. 138: 124-133.  https://doi.org/10.1016/j.jphotobiol.2014.05.010
  34. Song MW, Chung Y, Kim KT, Hong WS, Chang HJ, Paik HD. 2020. Probiotic characteristics of Lactobacillus brevis B13-2 isolated from kimchi and investigation of antioxidant and immune-modulating abilities of its heat-killed cells. LWT-Food Sci. Technol. 128: 109452. 
  35. Deng W, Long Q, Zeng J, Li P, Yang W, Chen X, Xie J. 2017. Mycobacterium tuberculosis PE_PGRS41 enhances the intracellular survival of M. smegmatis within macrophages via blocking innate immunity and inhibition of host defense. Sci. Rep. 7: 1-13.  https://doi.org/10.1038/s41598-016-0028-x
  36. Jeong M, Kim JH, Yang H, Dal Kang S, Song S, Lee D, Lee JS, Park JHY, Byun S, Lee KW. 2019. Heat-killed Lactobacillus plantarum KCTC 13314BP enhances phagocytic activity and immunomodulatory effects via activation of MAPK and STAT3 pathways. J. Microbiol. Biotechnol. 29: 1248-1254.  https://doi.org/10.4014/jmb.1905.05066
  37. Altan-Bonnet G, Mukherjee R 2019. Cytokine-mediated communication: a quantitative appraisal of immune complexity. Nat. Rev. Immunol. 19: 205-217.  https://doi.org/10.1038/s41577-019-0131-x
  38. Jeong DY, Lee ET, Lee J, Shin DC, Lee YH, Park JK. 2023. Effect of chemical structural properties of chitooligosaccharides on the immune activity of macrophages. Macromol. Res. 31: 569-582.  https://doi.org/10.1007/s13233-023-00143-8
  39. Sun X, Wang Z, Shao C, Yu J, Liu H, Chen H, et al. 2021. Analysis of chicken macrophage functions and gene expressions following infectious bronchitis virus M41 infection. Vet. Res. 52: 14. 
  40. Kim JG, Dong X, Park SH, Bayazid AB, Jeoung SA, Lim BO. 2021. Bioconversion of black rice and blueberry regulate immunity system through regulation of MAPKs, NF-kB in RAW264.7 macrophage cells. Food Agric. Immunol. 32: 471-481.  https://doi.org/10.1080/09540105.2021.1956434
  41. Vargas AM, Rivera-Rodriguez DE, Martinez LR. 2020. Methamphetamine alters the TLR4 signaling pathway, NF-κB activation, and pro-inflammatory cytokine production in LPS-challenged NR-9460 microglia-like cells. Mol. Immunol. 121: 159-166.  https://doi.org/10.1016/j.molimm.2020.03.013
  42. Yujiao H, Xinyu T, Xue F, Zhe L, Lin P, Guangliang S, et al. 2023. Selenium deficiency increased duodenal permeability and decreased expression of antimicrobial peptides by activating ROS/NF-κB signal pathway in chickens. BioMetals 36: 137-152. 
  43. Lee J, Kim S, Kang CH. 2022. Immunostimulatory activity of lactic acid bacteria cell-free supernatants through the activation of NF-κB and MAPK signaling pathways in RAW 264.7 cells. Microorganisms 10: 2247. 
  44. Zhou Y, Takano T, Li X, Wang Y, Wang R, Zhu Z, et al. 2022. β-Elemene regulates M1-M2 macrophage balance through the ERK/JNK/P38 MAPK signaling pathway. Commun. Biol. 5: 519.