[논문]할미밀망 에탄올 추출물의 항산화 및 항염증 활성 평가

정재미; 신미준; 정나은; 황다현

doi:10.15324/kjcls.2021.53.2.165

할미밀망 에탄올 추출물의 항산화 및 항염증 활성 평가
Antioxidant and Anti-Inflammatory Activities of Ethanol Extract of Clematis trichotoma Nakai 원문보기

Korean journal of clinical laboratory science : KJCLS = 대한임상검사과학회지, v.53 no.2, 2021년, pp.165 - 173

정재미 (호서대학교 생명보건대학 임상병리학과) , 신미준 (호서대학교 생명보건대학 임상병리학과) , 정나은 (호서대학교 생명보건대학 임상병리학과) , 황다현 (호서대학교 생명보건대학 임상병리학과)

초록
AI-Helper

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.

Abstract ▼ AI-Helper

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.

주제어

표/그림 (6)

표 Table 1. PCR primer lists
표 Table 2. Antioxidant activities of Clematis trichotoma Nakai
그림 Figure 2. Effect of CTN on dendritic transformation in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 31.25, 62.5, 125, 250 μg/mL of CTN for 30 min and co-treated by LPS for 24 h. NC was treated with only the culture medium, and the LPS group was treated with only 1 μg/mL LPS. Cell morphology was monitored under a light microscope at ×400 magnitude. Activated RAW 264.7 cells are indicated by arrows. The activation index percentage was expressed as the number of cells with activated morphology relative to the total number of cells, quantified in 5 random fields. Results are expressed as mean±SD of three independent tests in triplicate. Asterisks mean a significant difference between LPS group and each group by Student's t-test. **P<0.01, ***P<0.001. Abbreviations: See Figure 1.; ns, not significant.
그림 Figure 1. Cytotoxic effect of CTN in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 31.25, 62.5, 125, 250 μg/mL of CTN for 30 min and co-treated by LPS for 24 h. NC was treated with only the culture medium, and the LPS group was treated with only 1 μg/mL LPS. Results are expressed as mean±SD of three independent tests in triplicate. Asterisks mean a significant difference between LPS group and each group by Student's t-test. *P<0.05, **P<0.01, ***P<0.001. Abbreviations: NC, Negative control; LPS, Lipopolysaccharides; CTN, Clematis trichotoma Nakai.
그림 Figure 3. Anti-inflammatory activity of CTN in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 31.25, 62.5, 125, 250 μg/mL of CTN for 30 min and co-treated by LPS for 24 h. Supernatants were collected and (A) NO, (B) IL-6, (C) TNF-α were measured. NC was treated with only the culture medium, and the LPS group was treated with only 1 μg/mL LPS. Results are expressed as mean±SD of three independent tests in triplicate. Asterisks mean a significant difference between LPS group and each group by Student's t-test. *P<0.05, **P<0.01, ***P<0.001. Abbreviations: See Figure 1.
그림 Figure 4. Changes in the mRNA expression level of inflammatory mediators by CTN in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells were treated with 31.25, 62.5, 125, 250 μg/mL of CTN for 30 min and co-treated by LPS for 24 h. The cells were collected and (A) COX2, (B) iNOS, (C) IL-6, (D) TNF-α mRNA level were estimated NC was treated with only the culture medium, and the LPS group was treated with only 1 μg/mL LPS. Results are expressed as mean±SD of three independent tests in triplicate. Asterisks mean a significant difference between LPS group and each group by Student's t-test. *P<0.05, **P<0.01, ***P<0.001. Abbreviations: See Figure 2.

참고문헌 (29)

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?bmoderead&bSeq&aSeq385629&pageNo1&rowNum10&navCount10&currPg&searchInfo&sTargettitle&sTxt
Sung BK, Chung HY. Significances of molecular inflammation and energy metabolism during aging. J Cancer Prev. 2005;10: 6-17.
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

원문보기 상세보기
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.
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

원문보기 상세보기
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
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

원문보기 상세보기
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

상세보기
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

원문보기 상세보기
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.
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

원문보기 상세보기
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

상세보기
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.

원문보기 상세보기
Folin O, Denis WA. Colorimetric method for the determination of phenols (and phenol derivatives) in urine. J Biol Chem. 1915; 22:305-308.

상세보기
Stankovic MS. Total phenolic content, flavonoid concentration and antioxidant activity of Marrubium peregrinum L. extracts. Kragujevac J. Sci. 2011:33:63-72.
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.

상세보기
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.

상세보기
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

상세보기
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

상세보기
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.

상세보기
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

상세보기
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

상세보기
Maxwell SRJ. Prospects for the use of antioxidant therapies. Drugs. 1995;49:345-361.

상세보기
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

원문보기 상세보기
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

상세보기
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
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

원문보기 상세보기
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.
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.

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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