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Inhibitory Effect of Kailan (Brassica oleracea L.) Extract on LPS-Induced Inflammatory Response in Macrophages

카이란(Brassica oleracea L.) 추출물의 대식세포 내에서 LPS에 의한 염증 반응 억제 효과

  • DanHee Yoo (College of Fusion and Convergence, Seowon University) ;
  • In Chul Lee (Department of Bio-Cosmetic Science, Seowon University)
  • 유단희 (서원대학교 융복합대학) ;
  • 이인철 (서원대학교 바이오코스메틱학과)
  • Received : 2024.05.10
  • Accepted : 2024.07.25
  • Published : 2024.09.28

Abstract

In this study, antioxidant and anti-inflammatory activity was studied to confirm the value of kailan (Brassica oleracea L.) as a natural material for cosmetics. For this study measure the antioxidative activity, total polyphenol content was measured, and DPPH and ABTS scavenging activity assays were conducted. As a result of measuring the total polyphenol content of hot water extract of kailan (KRD) and 70% ethanol extract of kailan (KRE), it was found to contain 124.3 mg TAE/100 g and 144.1 mg TAE/100 g, respectively. As a result of DPPH and ABTS radical scavenging ability, it was confirmed that the efficacy was concentration dependent. After treating the cells with LPS, a stimulant, for 2 hours, an experiment was conducted by treating RAW 264.7 cells with KRD and KRE at concentrations of 10, 50, and 100 ㎍/ml. The nitric oxide production inhibitory activity of KRD and KRE showed an inhibitory effect of about 30% at a concentration of 100 ㎍/ml. Cells cultured for 18 hours after stimulant treatment were obtained and used in experiments. The cells obtained in this way were lysed, protein and mRNA were extracted, and the expression of inflammatory mediators' inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was confirmed. It was confirmed that the protein mRNA expression of iNOS and COX-2, measured through western blot and reverse transcription-PCR, was inhibited in a concentration-dependent manner. Based on this, it is judged that has the potential to be used as a natural material in cosmetics.

본 연구를 통해 카이란(Brassica oleracea L.)의 열수 추출물과 70% 에탄올 추출물의 항산화 및 항염 효과를 측정한 결과, 두 추출물 모두 항산화 및 항염에 효과를 가진 것으로 확인되었다. 카이란(Brassica oleracea L.)의 열수 추출물과 70% 에탄올 추출물의 실험 결과 총 폴리페놀 함량에서는 124.19 ± 0.19 mg/100 g, 144.10 ± 0.73 mg/100 g의 함량을 확인하였다. 열수 추출물과 70% 에탄올 추출물의 항산화 실험 결과 전자공여능 측정에서 1,000 ㎍/ml 농도에서 61.04%와 84.53%의 전자공여능이 나타나는 것을 확인하였고, ABTS 라디칼 소거능측정에서는 1,000 ㎍/ml 농도에서 89.13%와 98.61%의 소거능이 있는 것으로 확인하였다. 카이란의 열수 추출물과 70% 에탄올 추출물의 세포생존율 측정 결과, RAW 264.7 세포에서 100 ㎍/ml 농도에서 90% 이상의 세포생존율을 확인하였다. 카이란의 열수 추출물과 70% 에탄올 추출물의 NO 생성 저해 활성 측정 결과 100 ㎍/ml 농도에서 29.90%와 40.94%의 저해 활성을 확인하였다. LPS로 유도된 RAW 264.7 cell 대식세포에 대한 단백질 발현 억제 효과 측정 결과 카이란의 열수 추출물과 70% 에탄올 추출물의 COX-2의 억제 효과율은 100 ㎍/ml 농도에서 52.85%와 96.11%의 억제효과를 나타내었고 iNOS의 억제 효과율은 100 ㎍/ml 농도에서 12.13%와 18.73%의 억제율을 확인하였다. RT-PCR mRNA 측정 결과 카이란(Brassica oleracea L.)의 열수 추출물과 70% 에탄올 추출물의 COX-2에서 100 ㎍/ml 농도에서 98.16%와 92.74%의 발현이 측정되었고 iNOS에서는 92.74%와 76.86%의 발현이 측정되었다. 카이란 추출물의 실험 결과 두 추출물 모두 ABTS 라디칼 소거능 실험에서 대조군인 Vit. C 보다 좋은 효과를 보여 항산화능을 확인하였고 염증 발현 인자인 iNOS와 COX-2에 대해 단백질 발현 억제와 mRNA 발현에 대해 우수한 효능을 확인하였다. 위와 같은 결과들을 토대로, 카이란 추출물이 항산화, 항염 효과를 가진 것을 확인하여 향후 항산화 및 항염 천연 소재로 활용 가능성 및 가치를 가진 것으로 사료된다.

Keywords

References

  1. Yeom HY, Oh MJ, Chae JW, Lee JY. 2022. A study of anti-wrinkle activities as a functional cosmetic ingredient of Rhododendron brachycarpum extracts. J. Life Sci. 32: 622-632. 
  2. Anand SP, Sati N. 2013. Artificial preservatives and their harmful effects: looking toward nature for safer alternatives. Int. J. Pharm. Sci. Res. 4: 2496-2501. 
  3. Willoughby DA. 1975. Heberden Oration, 1974. Human arthritis applied to animal models. Towards a better therapy. Ann. Rheum. Dis. 34: 471-478. 
  4. Isomaki P, Punnonen J. 1997. Pro-and anti-inflammatory cytokines in rheumatoid arthritis. Ann. Med. 29: 499-507. 
  5. Kim SG. 2017. Antioxidant and anti-inflammatory activities of extracts from Ledum palustre L. Korean J. Food Preserv. 24: 1025-1033. 
  6. Fuentes F, Paredes-Gonzalez X, Kong ANT. 2015. Dietary glucosinolates sulforaphane, phenethyl isothiocyanate, indole-3-carbinol/3,3'-diindolylmethane: Antioxidative stress/inflammation, Nrf2, epigenetics/epigenomics and in vivo cancer chemopreventive efficacy. Curr. Pharmacol. Rep. 1: 179-196. 
  7. Singleton VL, Orthofer R, Lamuela-Raventos RM. 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Meth Enzymol. 299: 152-178. 
  8. Blois MS. 1958. Antioxidant determinations by the use or a stable free radical. Nature 181: 1199-1200. 
  9. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Rad. Biol. Med. 26: 1231-1237. 
  10. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. 1987. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47: 936-942. 
  11. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. 1982. Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal. Biochem. 126: 131-138. 
  12. Lee BY, Ahn JJ. 2022. Antioxidative and antimicrobial activities of Rhodiola rosea root as cosmetic material. Asian J. Beauty Cosmetol. 20: 541-549. 
  13. Chandrasekharan NV, Simmons DL. 2004. The cyclooxygenases. Genome Biol. 5: 241. 
  14. Song WY, Chun SS, Choi JH. 2018. Antioxidant activities of selenium-treated Spinacia oleracea L. J. Food Hyg. Saf. 33: 510-515. 
  15. Droge W. 2002. Free radicals in the physiological control of cell function. Physiol. Rev. 82: 47-95. 
  16. Jeong SJ, Kim KH, Yook HS. 2015. Whitening and antioxidant activities of solvent extracts. J. Korean Soc. Food Sci. Nutr. 44: 832-839. 
  17. Mo JH, Oh SJ. 2011. A study on P. Rhizoma extract's anti-micobial activity and cytotoxicity. Asian J. Beauty Cosmetol. 9: 1-12. 
  18. Lim CY. 2010. Nitric oxide and cancer. Korean J. Med. 78: 430-436. 
  19. Lee HJ, Park SE, Choi JH, Kim KM. 2022. Biological activity of Brassica oleracea var. capitata fermented with Lactobacillus plantarum. Korean J. Food Preserv. 29: 241-253. 
  20. Jeong DH, Kang BK, Kim KBWR, Kim MJ, Ahn DH. 2014. Anti-inflammatory activity of Sargassum micracanthum water extract. J. Appl. Biol. Chem. 57: 227-234. 
  21. Korbecki J, Baranowska-Bosiacka I, Gutowska I, Chlubek D. 2014. Cyclooxygenase pathways. Acta Biochim. Pol. 61: 639-649.