Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Enhancement of the Anti-inflammatory Activities of Aralia continentalis Kitagawa Extracts Fermented by Lactobacillus plantarum

추출용액에 따른 유산균 발효 땅두릅의 항염증 효과

  • Woo, Young Min (Natural Science Institute, Silla University) ;
  • Kim, Ok Ju (Natural Science Institute, Silla University) ;
  • Jo, Eun Sol (Natural Science Institute, Silla University) ;
  • Jo, Min Young (Department of Pharmaceutical Engineering, Silla University) ;
  • Ahn, Mee Young (Department of Pharmaceutical Engineering, Silla University) ;
  • Lee, Sang-Hyeon (Department of Pharmaceutical Engineering, Silla University) ;
  • Ha, Jong-Myung (Department of Pharmaceutical Engineering, Silla University) ;
  • Kim, Andre (Department of Pharmaceutical Engineering, Silla University)
  • 우영민 (신라대학교 자연과학연구소) ;
  • 김옥주 (신라대학교 자연과학연구소) ;
  • 조은솔 (신라대학교 자연과학연구소) ;
  • 조민영 (신라대학교 바이오산업학부 제약공학전공) ;
  • 안미영 (신라대학교 바이오산업학부 제약공학전공) ;
  • 이상현 (신라대학교 바이오산업학부 제약공학전공) ;
  • 하종명 (신라대학교 바이오산업학부 제약공학전공) ;
  • 김안드레 (신라대학교 바이오산업학부 제약공학전공)
  • Received : 2018.10.22
  • Accepted : 2018.11.08
  • Published : 2018.12.30
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Abstract

We investigated the anti-inflammatory activities of various organic solvent extracts with and without Lactobacillus plantarum fermentation of Aralia continentalis Kitagawa which has hypotensive effects in addition to excitatory effects on the central nervous system. It has been used to treat arthritis, colds, neuralgia, rheumatism, and itchy skin. Our extracts were tested for their anti-inflammatory potential on NO production and the expression of inflammatory factors in lipopolysaccharide-stimulated RAW264.7 macrophages. Extracts with and without L. plantarum fermentation were prepared using water, ethanol, hexane, ethyl acetate, and butanol. The RAW264.7 cells were tested for toxicity and the anti-inflammatory activity of each extract was determined at a concentration with no toxicity to the cells. The extracts used in this study significantly inhibited both the production of NO and the mRNA expression of COX-2 and iNOS, the major inflammatory factors. The production of inflammation-related cytokines $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ was also significantly reduced. These results suggest that the extracts involving fermentation by L. plantarum can inhibit cytokines by controlling the expression of inflammatory cytokine genes. It is considered that the water, ethanol, and butanol extracts after fermentation with L. plantarum could be useful as functional natural materials with anti-inflammatory effects.

본 연구는 추출 용매별 땅두릅 추출물과 Lactobacillus plantarum으로 발효한 후 용매별로 추출한 땅두릅 발효추출물을 이용하여 항염증 효과를 측정한 연구이다. 땅두릅은 중추신경 계통에 대한 흥분작용이 있고 혈압강하 작용이 알려져 있으며 관절염, 감기, 신경통, 류마티스, 피부가려움증 등에 쓰인다. LPS (lipopolysaccharide)로 염증을 유도한 마우스 유래 macrophage에서의 NO 생성 및 염증관련인자의 발현에 미치는 영향을 검토하여 항염증 소재로서의 이용 가능성을 확인하였다. 건조 땅두릅을 water, ethanol, hexane, ethyl acetate, butanol을 이용하여 추출한 추출물들과 L. plantarum으로 발효한 후 추출한 추출물들에서 RAW264.7 대식세포에 대한 독성 여부를 측정하였고, 세포에 대한 독성이 나타나지 않는 농도에서 추출물의 항염증 활성을 확인하였다. 본 실험에 사용된 추출물은 LPS로 유도된 NO 생성을 유의적으로 억제하였으며, 주요 염증 유발인자인 COX-2와 iNOS의 발현 또한 유의적으로 억제하는 효과를 나타냈다. 염증관련 cytokine인 $IL-1{\beta}$, IL-6 및 $TNF-{\alpha}$의 생성량 또한 유의적으로 감소시켰다. 이상의 결과로부터 땅두릅을 L. plantarum으로 발효한 후 water, ethanol, butanol로 추출하였을 때 염증 억제 효과가 있는 기능성 식품 소재로서의 개발 가능성이 있을 것으로 사료된다.

Keywords

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Fig. 1. Effect of various extracts from A. continentalis Kitagawa on the viability of the RAW264.7 cells by the CCK assay.

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Fig. 2. Effect of various extracts from A. continentalis Kitagawa on LPS-induced nitrite production in RAW264.7 cells.

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Fig. 3. Effect of various extracts from A. continentalis Kitagawa on COX-2 and iNOS gene expression in the LPS-induced RAW264.7 cells.

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Fig. 4. Effect of various extracts from A. continentalis Kitagawa on the production and gene expression of inflammatory cytokines in the LPS-induced RAW264.7 cells.

Table 1. Primers for RT-PCR analysis

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Table 2. Effect of various extracts from A. continentalis Kitagawa on DPPH radical scavenging activity

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References

  1. Akihisa, T., Kokke, W. C. M. C., Kimura, Y. and Tamura, T. 1993. Isokarounidiol (D-C-Friedooleana-6,8-diene-3-alpha, 29-diol) the first naturally occurring triterpene with a delta-6,8-conjugated diene system-Iodine-mediated dehydrogenation and isomerization of its diacetate. J. Org. Chem. 58, 1959-1962. https://doi.org/10.1021/jo00059a063
  2. An, S. M., Kim, H. G., Choi, E. J., Hwang, H. H., Lee, E. S., Beak, J. H., Bhu, Y. C. and Ko, J. S. 2014. Screening for antiinflammatory activities in extracts from Korean herb medicines. J. Soc. Cosmetic Sci. Korea 40, 95-108.
  3. Aono, K., Isobe, K., Kiuchi, K., Fan, Z. H., Ito, M., Takeuchi, A., Miyachi, M., Nakashima, I. and Nimura, Y. 1997. In vitro and in vivo expression of inducible nitric oxide synthase during experimental endotoxemia: involvement of other cytokines. J. Cell. Biochem. 65, 349-358. https://doi.org/10.1002/(SICI)1097-4644(19970601)65:3<349::AID-JCB5>3.0.CO;2-S
  4. Blois, M. S. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181, 1198-1200.
  5. Cha, J. H., Kim, Y. S. and Lee, E. M. 2010. Effects of Prunellae Spica water extract on immune response in macrophage cells. J. Orien. Obstet. Gynecol. 23, 91-100.
  6. Choi, W. S., Kwon, H. S., No, R. H., Choi, G. P. and Lee, H. Y. 2013. Enhancement of anti-inflammatory activities of fermented Scutellaria baicalensis extracts using Lactobacillus rhamnosus. J. Soc. Cosmet. Scientists Korea 39, 303-311. https://doi.org/10.15230/SCSK.2013.39.4.303
  7. Dang, N. H., Zhang, X. F., Zheng, M. S., Son, K. H., Chang, H. W., Kim, H. P., Bae, K. H. and Kang, S. S. 2005. Inhibitory constituents against cyclooxygenases from Aralia cordata Thunb. Arch. Pharm. Res. 28, 28-33. https://doi.org/10.1007/BF02975131
  8. Ha, Y. B., Park. J. H., Jang. J. W., Lim, D. W. and Kim, J. E. 2016. Comparative study for anti-inflammatory and anti-obesity effect of fractions from leaf and stem of Sasa borealis. J. Physiol. Pathol. Kor. Med. 30, 229-235. https://doi.org/10.15188/kjopp.2016.08.30.4.229
  9. Han, W. S. 2005. Isolation of the antimicrobial compounds from Aralia cordata Thunb. extreact. Kor. J. Medicinal Crop. Sci. 13, 182-185.
  10. Han, W., Mercenier, A., Belgnaoui, A. A., Pavan, S., Lamine, F., Swan, I. I., Kleerebezem, M., Cartier, C. S., Hisbergues, M., Bueno, L., Theodorou, V. and Fioramonti, J. 2006. Improvement of an experimental colitis in rats by lactic acid bacteria producing superoxide dismutase. Inflamm. Bowel Dis. 12, 1044-1052. https://doi.org/10.1097/01.mib.0000235101.09231.9e
  11. Jeong, H., Kim, H., Ju, E., Lee, S. G., Kong, C. S. and Seo, Y. 2017. Antiinflammatory activity of solvent-partitioned fractions from Atriplex gmelinii C. A. Mey. in LPS-stimulated RAW264.7 macrophages. J. Life. Sci. 2, 187-193.
  12. Jeong, S. I., Han, W. S., Yun, Y. H. and Kim, K. J. 2006. Continentalic acid from Aralia continentalis shows activity against methicillin-resistant Staphylococcus aureus. Phytother. Res. 20, 511-514. https://doi.org/10.1002/ptr.1894
  13. Jin, K. S., Lee, J. Y., Kwon, H. J. and Kim, B. W. 2014. Antioxidative and anti-inflammatory activities of Ardisia arborescens ethanol extract. J. Life Sci. 24, 713-720. https://doi.org/10.5352/JLS.2014.24.7.713
  14. Kang, C. H., Koo, J. R. and So, J. S. 2012. Inhibitory effects of Aralia cordata Thumb extracts on nitric oxide synthesis in RAW 264.7 macrophage cells. Kor. J. Food Sci. Technol. 44, 621-627. https://doi.org/10.9721/KJFST.2012.44.5.621
  15. Kang, M. S., Oh, H. J., Lee, H. C. and Oh, J. S. 2009. Isolation and identification of lactic acid bacteria inhibiting the proliferation of Propionibacterium acnes and Staphylococcus epidermidis. J. Bacteriol. Virol. 39, 11-19. https://doi.org/10.4167/jbv.2009.39.1.11
  16. Kim, C. M. and Park, Y. K. 2009. The effects of different extracts of Ostericum koreanum on the production of inflammatory mediators in LPS-stimulated RAW264.7 cells. Kor. J. Herbology 24, 169-178.
  17. Kim, H. S. and Ham, J. S. 2003. Antioxidative ability of lactic acid bacteria. Kor. J. Food Sci. Ani. Resoul. 23, 186-192.
  18. Kim, P. K., Jung, K. I., Choi, Y. J. and Gal, S. W. 2017. Anti-inflammatory effects of lemon myrtle (Backhousia citriodora) leaf extracts in LPS-induced RAW 264.7 Cells. J. Life Sci. 9, 986-993.
  19. Kim, S. I., Lee, S. M., Lee, C. Y., Son, H. J., Hwang, D. Y., Lee, H. and Kim, D. S. 2016. Antimicrobial activity and characteristics of Asparagus Cochinchinensis fermented with lactic acid bacteria. Food Eng. Prog. 20, 278-284. https://doi.org/10.13050/foodengprog.2016.20.4.278
  20. Kong, C. S. 2014. Anti-inflammatory activity of the solventpartitioned fractions from Spergularia marina in LPS stimulated RAW 264.7 cells. Prev. Nutr. Food Sci. 19, 261-267. https://doi.org/10.3746/pnf.2014.19.4.261
  21. Lee, H., Yang, S. G., Park, S. N. and Jeon, D. Y. 1992. Effect of Lactobacilli on reactive oxygen scavenging and immune stimulation. Kor. J. Biotechnol. Bioeng. 7, 290-295.
  22. Lee, K. H. and Rhee, K. H. 2015. Screening of anti-Inflammatory herbs having the activation of MAPK family proteins. Kor. J. Food Nutr. 28, 343-350. https://doi.org/10.9799/ksfan.2015.28.3.343
  23. Lee, S. E., Seong, N. S., Bang, J. K., Park, C. G., Sung, J. S. and Song, J. 2003. Antioxidative activities of Korean medicinal plants. Kor. J. Med. Crop Sci. 11, 127-134.
  24. Lee, S. G., Jo, D. J., Chang, H. J. and Kang, H. 2015. Antioxidant and anti-inflammatory activities of ethanol extracts from Aralia continentalis Kitagawa. J. Naturopath. 4, 10-14.
  25. Lim, S. M. 2010. Resistance to reactive oxygen species and antioxidant activities of some strains of lactic acid bacteria from the mustard leaf Kimchi. Kor. J. Microbiol. 46, 375-382.
  26. Masood, M. I., Qadir, M. I., Shirazi, J. H. and Khan, I. U. 2011. Beneficial effects of lactic acid bacteria on human beings. Crit. Rev. Microbiol. 37, 91-98. https://doi.org/10.3109/1040841X.2010.536522
  27. Moncade, S. and Higges, A. 1993. The L-arginine-nitric oxide pathway. N. Engl. J. Med. 329, 2002-2012. https://doi.org/10.1056/NEJM199312303292706
  28. Mun, O. J., Kwon, M. S., Bae, M. J., Ahn, B. N., Fatih, K., Kim, M. H., Lee, S. H., Yu, K. H., Kim, Y. Y., Seo, Y. W. and Kong, C. S. 2015. Anti-inflammatory activity of Hizikia fusiformis extracts fermented with Lactobacillus casei in LPS stimulated RAW 264.7 macrophages. Kor. Soc. Biotechnol. Bioeng. J. 30, 38-43.
  29. Padwad, Y., Ganju, L., Jain, M., Chanda, S., Karan, D., Banerjee, P. K. and Sawhney, R. C. 2006. Effect of leaf extract of Seabuckthorn on lipopolysaccharide induced inflammatory response in murine macrophages. Int. Immunopharmacol. 6, 46-52. https://doi.org/10.1016/j.intimp.2005.07.015
  30. Park, H. J., Hong, M. S., Lee, J. S., Leem, K. H., Kim, C. J., Kim, J. W. and Lim, S. 2005. Effects of aralia continentalis on hyperalgesia with peripheral inflammation. Phytother. Res. 19, 511-513. https://doi.org/10.1002/ptr.1693
  31. Reddy, G. V., Friend, B. A., Shahani, K. M. and Farmer, R. E. 1983. Antitumor activity of yogurt components. J. Food Prot. 46, 8-11. https://doi.org/10.4315/0362-028X-46.1.8
  32. Ryu, H. S. 2008. Effects of Job's Tears (Yul-Moo) extracts on mouse splenocyte and macrophage cell activation. Kor. J. Food Nutr. 21, 1-6.
  33. Seok, E. K., Kim, T. H., Lee, J. C., Chung, P. K. and Lee, K. K. 1987. Lowering of serum cholesterol by Lactobacillus sporogenes. Yakhak Hoeji 31, 302-307.
  34. Shahani, K. M., Friend, B. A. and Bailey, P. J. 1983. Antitumor activity of fermented colostrum and milk. J. Food Prot. 46, 385-386. https://doi.org/10.4315/0362-028X-46.5.385
  35. Stuehr, D. J., Cho, H. J., Kwon, N. S., Weise, M. F. and Nathan, C. F. 1991. Purification and characterization of the cytokine-induced macrophage nitric oxide synthase: An FAD- and FMN-containing flavoprotein. Proc. Natl. Acad. Sic. USA. 88, 7773-7777. https://doi.org/10.1073/pnas.88.17.7773
  36. Turini, M. E. and DuBois, R. N. 2002. Cyclooxygenase-2: a therapeutic target. Annu. Rev. Med. 53, 35-57. https://doi.org/10.1146/annurev.med.53.082901.103952
  37. Yang, H. M., Lim, S. S., Lee, Y. S., Shin, H. K., Oh, Y. S. and Kim, J. K. 2007. Comparison of the anti-inflammatory effects of the extracts from Rubus coreanus and Rubus occidentalis. Kor. J. Food Sci. Technol. 39, 342-347.
  38. Yang, S. A., Im, N. K., Jhee, K. H. and Lee, I. S. 2008. Effects of Aralia continentalis Kitagawa on antiplatelet and antioxidative activities. J. Life Sci. 18, 357-362. https://doi.org/10.5352/JLS.2008.18.3.357
  39. Yu, A R., Park, H. Y., Choi, I. W., Park, Y. K., Hong, H. D. and Choi, H. D. 2012. Immune enhancing effect of medicinal herb extracts on a RAW 264.7 macrophage cell line. J. Kor. Soc. Food Sci. Nutr. 41, 1521-1527. https://doi.org/10.3746/jkfn.2012.41.11.1521
  40. Yun, S. G., Jin, S., Jeong, H. Y., Yun, H. J., Do, M. Y., Kim, B. W. and Kwon, H. J. 2015. Anti-oxidant, anti-inflammatory and anti-cancer effect of methanol extract of Pogostemon cablin. J. Life Sci. 25, 44-52. https://doi.org/10.5352/JLS.2015.25.1.44