Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Antioxidant and Anti-Inflammatory Activities of Ethanol Extract of Clematis trichotoma Nakai

할미밀망 에탄올 추출물의 항산화 및 항염증 활성 평가

  • Jung, Jaemee (Department of Biomedical Laboratory Science, College of Life and Health Sciences, Hoseo University) ;
  • Shin, Mijoon (Department of Biomedical Laboratory Science, College of Life and Health Sciences, Hoseo University) ;
  • Jeong, Naeun (Department of Biomedical Laboratory Science, College of Life and Health Sciences, Hoseo University) ;
  • Hwang, Dahyun (Department of Biomedical Laboratory Science, College of Life and Health Sciences, Hoseo University)
  • 정재미 (호서대학교 생명보건대학 임상병리학과) ;
  • 신미준 (호서대학교 생명보건대학 임상병리학과) ;
  • 정나은 (호서대학교 생명보건대학 임상병리학과) ;
  • 황다현 (호서대학교 생명보건대학 임상병리학과)
  • Received : 2021.05.21
  • Accepted : 2021.05.26
  • Published : 2021.06.30
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Abstract

Clematis trichotoma Nakai (CTN) is a broad-leaved vine plant belonging to the family Ranunculus, native to Korea. Young leaves are used as food, and the stem and roots are used as medicinal materials. Antioxidant studies have been reported on the stems of CTN, but no studies have been conducted on the leaves. In this study, a 70% ethanol extract of CTN was prepared and its antioxidant and anti-inflammatory activities were investigated. For measuring the antioxidant activity, five assays (polyphenol and flavonoid content, reducing power, 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) were performed and CTN showed a concentration-dependent effect in all assays. To investigate the anti-inflammatory activity, we used RAW 264.7 cells. The concentrations (from 31.25 to 250 ㎍/mL) of CTN did not show cytotoxicity. CTN (250 ㎍/mL) inhibited dendritic transformation (34.4%) and also inhibited inflammation as seen by reduced levels of NO (77.4%), IL-6 (85.5%) and TNF-α (41.2%) compared to lipopolysaccharide (LPS). CTN (250 ㎍/mL) also suppressed the expression of the following genes: COX-2 (79.8%), iNOS2 (93.9%), IL-6 (87.6%), and TNF-α (77.3%) compared to LPS. These results demonstrated that CTN has excellent antioxidant and anti-inflammatory activities and can therefore be used as a natural biological resource.

할미밀망은 국내에서 자생하는 미나리아재비과의 낙엽활엽덩굴식물이다. 어린 잎은 식용으로 사용되며 줄기와 뿌리는 약재로 사용된다. 할미밀망 줄기에 대한 항산화 연구는 보고되었으나 잎에 대한 연구는 현재 수행되지 않았다. 본 연구에서는 할미밀망을 70% 에탄올 추출물로 제조하고 항산화 및 항염증 생리 활성을 비교하였다. 항산화 활성을 측정하기 위해 5가지 분석(총 폴리 페놀 함량, 플라보노이드 함량, 환원력, ABTS 및 DPPH 라디칼 소거 활성)이 사용되었으며 모든 분석에서 농도 의존적 효과를 나타냈다. 항염증 활성을 조사하기 위해 RAW 264.7 세포를 사용했다. 할미밀망은 농도(31.25-250 ㎍/mL)에서 세포 독성을 나타내지 않았다. 할미밀망(250 ㎍/mL)은 LPS에 비해 세포의 수지상적 형태 변화를 억제하였고(34.4%), NO (77.4%), IL-6 (85.5%) 및 TNF-α (41.2%)의 발현을 억제함으로써 염증을 낮추는 효과를 나타내었다. 또한 할미밀망(250 ㎍/mL)은 LPS 단독 처리군에 비해 COX-2 (79.8%), iNOS2 (93.9%), IL-6 (87.6%) 및 TNF-α (77.3%)의 유전자 발현을 억제했다. 이러한 결과는 할미밀망이 우수한 항산화 및 항염증 작용을 가지고 있음을 보여준다. 그러므로 할미밀망은 향후 항산화 및 항염증 효과를 가진 천연 생물 유래 소재로 활용될 수 있을 것이다.

Keywords

Acknowledgement

This research was supported by the Academic Research Fund of Hoseo University in 2020-0431.

References

  1. Statistics Korea. Causes of death statistics in 2019 [Internet]. Seoul: Statistics Korea; 2020. 9. 22. [cited 2021 May 1]. Available from:http://kostat.go.kr/portal/eng/pressReleases/8/10/index.-board?bmode=read&bSeq=&aSeq=385629&pageNo=1&rowNum=10&navCount=10&currPg=&searchInfo=&sTarget=title&sTxt=
  2. Sung BK, Chung HY. Significances of molecular inflammation and energy metabolism during aging. J Cancer Prev. 2005;10: 6-17.
  3. Lee SG, Kang H. Effects of Houttuynia cordata extracts of different aerial parts on antioxidants and anti-inflammatory. Biomed Sci Lett. 2018;24:87-92. https://doi.org/10.15616/BSL.2018.24.2.87
  4. Han JS, Yi DH. Effects of pine needles fermentation extracts on antioxidant activity and inhibition of melanin synthesis. Asian J Beauty Cosmetol. 2012;10:619-624.
  5. Leem HH, Kim EO, Seo MJ, Choi SW. Antioxidant and anti-inflammatory activities of Eugenol and its derivatives from Clove (Eugenia caryophyllata Thunb.). J Korea Soc Food Sci Nutr. 2011;40:1361-1369. https://doi.org/10.3746/jkfn.2011.40.10.1361
  6. Na EJ, Jang HH, Kim GR. Review of recent studies and research analysis for anti-oxidant and anti-aging materials. J Korea Soc Food Sci Nutr. 2016;14:481-491. https://doi.org/10.20402/ajbc.2016.0107
  7. Park GH, Lee JY, Kim DH, Cho YJ, An BJ. Anti-oxidant and anti-inflammatory effects of Rosa multiflora root. J Life Sci. 2011;21: 1120-1126. https://doi.org/10.5352/JLS.2011.21.8.1120
  8. Kim KS, Ryu MJ. Physiological activity of the Glycyrrhiza uralensis extracts as a cosmetic product. Asian J Beauty Cosmetol. 2017;15:11-22. https://doi.org/10.20402/ajbc.2016.0084
  9. Cheon YP, Mohammad LM, Park CH, Hong JH, Lee GD, Song JC, et al. Bulnesia sarmienti aqueous extract inhibits inflammation in LPS-stimulated RAW 264.7 Cells. J Life Sci. 2009;19:479-485. https://doi.org/10.5352/JLS.2009.19.4.479
  10. Cho YJ, An BJ. Anti-inflammatory effect of extracts from Cheongmoknosang (morus alba L.) in lipopolysaccharide-stimulated Raw cells. Appl Biol Chem. 2008;51:44-48.
  11. Kim DH, Hwang EY, Son JH. Anti-inflammatory activity of Carthamus tinctorious seed extracts in Raw 264.7 cells. J Life Sci. 2013;23:55-62. https://doi.org/10.5352/JLS.2013.23.1.55
  12. Kim H, Kim TH, Jeon BR, Bang MH, Kim WJ, Park JW, et al. Evaluation of the physiological activity on the skin and identification of the active ingredient of leaf extract from Sanhyang sandolbae (Pyrus ussuriensis) as a new variety. J Korean Soc Food Sci Nutr. 2020;49:35-44. https://doi.org/10.3746/jkfn.2020.49.1.35
  13. Tran MH, Phuong TT, Ki HB. Antioxidant effect of flavonoids isolated from the root of Clematis trichotoma Nakai. Korean J Medicinal Crop Sci. 2005;13:227-231.
  14. Folin O, Denis WA. Colorimetric method for the determination of phenols (and phenol derivatives) in urine. J Biol Chem. 1915; 22:305-308. https://doi.org/10.1016/S0021-9258(18)87648-7
  15. Stankovic MS. Total phenolic content, flavonoid concentration and antioxidant activity of Marrubium peregrinum L. extracts. Kragujevac J. Sci. 2011:33:63-72.
  16. Benzie IFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem. 1996;239:70-76. https://doi.org/10.1006/abio.1996.0292
  17. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26:1231-1237. https://doi.org/10.1016/S0891-5849(98)00315-3
  18. Cho BO, Che DN, Kim J, Kim JH, Shin JY, Kang HJ, et al. In vitro anti-inflammatory and anti-oxidative stress activities of Kushenol C isolated from the roots of Sophora flavescens. Molecules 2020;25:1768. https://doi.org/10.3390/molecules25081768
  19. Livak KJ and Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25:402-408. https://doi.org/10.1006/meth.2001.1262
  20. Saxena RK, Vallyathan V, Lewis DM. Evidence for lipopolysaccharide-induced differentiation of RAW264.7 murine macrophage cell line into dendritic like cells. J Biosci. 2003; 28:129-134. https://doi.org/10.1007/BF02970143
  21. Zin ZM, Abdul-Hamid A, Osman A. Antioxidative activity of extracts from Mengkudu (Morinda citrifolia L.) root, fruit and leaf. Food Chem. 2002;78:227-231. https://doi.org/10.1016/S0308-8146(01)00402-2
  22. Jubany-Mari Tana, Munne-Bosch Sergi, Lopex-Carbonell Marta, Leonor A. Hydrogen peroxide is involved in the acclimation of the Mediterranean shrub, Cistus albidus L., to summer drought. J Exp Bot. 2009;60:107. https://doi.org/10.1093/jxb/ern274
  23. Maxwell SRJ. Prospects for the use of antioxidant therapies. Drugs. 1995;49:345-361. https://doi.org/10.2165/00003495-199549030-00003
  24. Kim EJ, Choi JY, Yu MR, Kim MY, Lee SH, Lee BH. Total polyphenols, total flavonoid contents, and antioxidant activity of Korean natural and medicinal plants. Korean J Food Sci. Technol. 2012;44:337-342. https://doi.org/10.9721/KJFST.2012.44.3.337
  25. Yen G, Duh P, Tsai H. Antioxidant and pro-oxidant properties of ascorbic acid and gallic acid. Food Chem. 2002;79:307-313. https://doi.org/10.1016/S0308-8146(02)00145-0
  26. Kim MJ, Bae GS, Choi JW, Jo IJ, Kim DG, Shin JY, et al. The anti-inflammatory effect of Taraxacum coreanum on lipopolysaccharide induced inflammatory response on RAW 264.7 cells. Kor J Herbology. 2014;29:21-26. https://doi.org/10.6116/kjh.2014.29.6.21
  27. Bae NY, Kim MJ, Kim BYWR, Park JH, Park SH, Sung NK, et al. Anti-inflammatory effect of Chondrus ocellatus holmes ethanol extract on lipopolysaccharide-induced inflammatory responses in RAW 264.7 cells. J Microbial Biotechnol. 2016;44:268-277. https://doi.org/10.4014/mbl.1603.03004
  28. Jew SS, Bae ON, Chung JH. Anti-inflammatory effects of asiaticoside on inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cell line. Toxicol Res. 2003;19:33-37.
  29. Lee KH, Kim HY, Jung HJ, Lee HK. Effect of Chungsangboha-tang on LPS induced anti-inflammatory in THP-1 cells. Korean J. Orient Int. Med. 2008;29:12-24.