[논문]LPS에 의해 활성화된 대식세포에서 애플망고 껍질(Mangifera indica L. Peel)의 항염증 효능 검증

김효민; 유단희; 이인철

doi:10.48022/mbl.2207.07002

LPS에 의해 활성화된 대식세포에서 애플망고 껍질(Mangifera indica L. Peel)의 항염증 효능 검증
Verification of Anti-Inflammatory Efficacy of Apple Mango (Mangifera indica L.) Peel in LPS-Activated Macrophage 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.50 no.3, 2022년, pp.337 - 346

김효민 (경북대학교 약학대학) , 유단희 (서원대학교 융복합대학) , 이인철 (서원대학교 바이오코스메틱학과)

초록
AI-Helper

최근 국내 기후 변화로 인해 열대 과일 재배가 가능해지게 되었다. 특히 대표적으로 제주도에서 애플망고의 재배가 활발히 이루어지고 있으며 이로 인해 소비자들의 수요가 증가하고 있다. 그러나 애플망고 과육을 제외한 껍질과 같은 부산물은 활용도가 떨어져 폐기되고 있는 실정이다. 이에 본 연구에서는 부산물로서 폐기되어지는 애플망고 껍질을 활용하기 위해 이를 이용하여 항산화 및 항염증 효과를 검증하고자 열수 및 70% 에탄올로 추출하여 실험을 진행하였다. 먼저 애플망고 껍질의 열수 및 70% 에탄올 추출물의 총 폴리페놀 함량을 확인하였으며, 그 결과 66.08 ± 0.12 mg TAE/100 g, 100.13 ± 0.23 mg TAE/100 g의 함량을 나타내었다. 항산화 활성을 측정하기 위해 전자공여능과 2,2'- azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) 라디칼 소거능을 측정하였다. 전자공여능 측정결과, 1,000 ㎍/ml 농도에서 열수 추출물은 86.89%가 나타났고, 70% 에탄올 추출물은 94.06%를 확인하였다. ABTS 라디칼 소거능을 측정 결과로 1,000 ㎍/ml 농도에서 열수 추출물은 80.62% 를 나타내었고, 70% 에탄올 추출물은 98.66%의 소거능을 확인하였다. 이로 인해 애플망고 껍질 추출물의 항산화 활성은 대조군인 비타민 C와 유사한 효과를 가진 것을 확인하였다. 애플망고 껍질 추출물의 세포 생존율을 측정한 결과, 대식세포인 RAW 264.7 세포에서는 500 ㎍/ml 농도에서 99% 이상의 세포 생존율을 확인하였다. 이후 세포실험은 세포생존율이 99% 이상인 농도구간을 설정하여 진행하였다. 염증성 매개 물질로 알려진 nitric oxide (NO)에 대한 생성 억제 효과를 확인한 결과, 애플망고 껍질의 열수 추출물은 46%의 억제 효과가 나타났고, 70% 에탄올 추출물은 40%의 억제 효과를 확인하여 애플망고 추출물의 NO 생성량 저해 효과를 가진 것을 확인하였다. 염증 매개 인자의 억제 효과를 확인하기 위해 RAW 264.7 세포를 이용하여 iNOS와 COX-2의 단백질 및 mRNA 발현량을 측정하였다. 그 결과, LPS 단독 처리군에 비해 애플망고 껍질 추출물을 처리하였을 때 iNOS와 COX-2의 단백질 발현량은 최종 농도인 500 ㎍/ml에서 열수 추출물은 12%, 97% 억제하는 것을 확인하였고, 70% 에탄올 추출물은 62%, 91% 억제하는 것을 확인하였다. 또한 LPS 단독 처리군에 비해 iNOS와 COX-2의 mRNA 발현량 결과, 500 ㎍/ml 농도에서 열수 추출물은 3%, 33%의 억제효과를 확인하였고, 70% 에탄올 추출물은 29%, 36%의 억제효과가 나타나는 것을 확인하였다. 본 연구 결과들을 토대로 애플망고 껍질이 항산화 및 항염증 활성을 갖고 있는 것을 검증하여 천연 소재로서의 활용 가치가 높을 것으로 사료된다.

Abstract ▼ AI-Helper

The purpose of this study was to investigate the antioxidant and anti-inflammatory activities of hot water (AMPW) and 70% ethanol (AMPE) extracts of apple mango (Mangifera indica L.) peel. The antioxidant activities were measured using a total polyphenol, electron-donating, 2,2'-azinobis [3-ethylbenzothiazoline6-sulfonic acid] (ABTS) radical scavenging assay. The total polyphenol content of AMPW and AMPE was 66.08 ± 0.62 mg TAE/100 g and 100.13 ± 0.23 mg TAE/100 g, respectively. As a result of measuring the electrondonating ability, at a concentration of 1,000 ㎍/ml, AMPW and AMPE showed an effectiveness of 86% and 94%, respectively. The ABTS assay showed 80% and 98% respective radical scavenging activity for AMPW and AMPE, at a concentration of 1,000 ㎍/ml. The cell viability on macrophage cells was performed using a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay, and the results showed more than 90% cell viability at a 100 ㎍/ml concentration. Anti-inflammatory activity was verified by confirming nitric oxide (NO) production inhibitory activity, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) protein and mRNA expression inhibitory activity from lipopolysaccharide (LPS)-treated RAW 264.7 cells. The NO production inhibitory effects were measured using the Griess assay, which confirmed 45% and 40% inhibition after treatment with AMPW and AMPE, respectively. Moreover, the protein and mRNA expression of inflammatory-related factors iNOS and COX-2, decreased in a concentrationdependent manner. In conclusion, this study showed antioxidant and anti-inflammatory effects of Mangifera indica L. peel and revealed its promising potential for application as an antioxidant and anti-inflammatory agent.

주제어

표/그림 (10)

표 Table 1. Sequence of the primers used for RT-PCR.
그림 Fig. 1. Electron donating activity of extracts from Mangifera indica L. peel. ■ AMPW: Apple mango peel extracted with distilled water at 100℃ ▨ AMPE: Apple mango peel extracted with 70% ethanol at room temperature ☐ Vit. C: L-ascorbic acid Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs Vit. C)
표 Table 2. Total polyphenol contents in Mangifera indica L. peel.
그림 Fig. 2. ABTS radical scavenging activity of extracts from Mangifera indica L. peel. ■ AMPW: Apple mango peel extracted with distilled water at 100℃ ▨ AMPE: Apple mango peel extracted with 70% ethanol at room temperature ☐ Vit. C: L-ascorbic acid Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs Vit. C).
그림 Fig. 3. Cell viability of extracts from Mangifera indica L. peel on RAW 264.7 cells. In order to measure the cell viability using the MTT assay, Mangifera indica L. peel extracts were treated with each concentration (5, 10, 50, 100, 500 and 1,000 μg/ml) in RAW 264.7 cells. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs control). ■ AMPW: Apple mango peel extracted with distilled water at 100℃ ☐ AMPE: Apple mango peel extracted with 70% ethanol at room temperature
그림 Fig. 4. Inhibitory effects of extracts from Mangifera indica L. peel on NO production in LPS-induced RAW 264.7 cells. RAW 264.7 cells (2 × 10⁵ cells/well) were pretreated with LPS (10 μg/ml) for 2 h and then treated Mangifera indica L. peel extracts at an appropriate concentration (10, 50, 100 and 500 μg/ml) for 16 h. NO production was determined in culture supernatant by griess reagent. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.00 vs LPS alone-treated group). ■ AMPW: Apple mango peel extracted with distilled water at 100℃ ▨ AMPE: Apple mango peel extracted with 70% ethanol at room temperature
그림 Fig. 5. iNOS, COX-2 protein expression levels of hot water extracts from Mangifera indica L. peel in RAW 264.7 cells. RAW 264.7 cells (1 × 10⁶ cells/well) were pretreated with LPS (10 μg/ml) for 2 h and then treated water extracts from Mangifera indica L. peel at an appropriate concentration (10, 50, 100 and 500 μg/ml) for 16 h. iNOS and COX-2 protein expression was determined in RAW 264.7 cells treated with AMPW. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs LPS alone-treated group).
그림 Fig. 6. iNOS, COX-2 protein expression levels of 70% ethanol extracts from Mangifera indica L. peel in RAW 264.7 cells. RAW 264.7 cells (1 × 10⁶ cells/well) were pretreated with LPS (10 μg/ml) for 2 h and then treated ethanol extracts from Mangifera indica L. peel at an appropriate concentration (10, 50, 100 and 500 μg/ml) for 16 h. iNOS and COX-2 protein expression was determined in RAW 264.7 cells treated with AMPE. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs LPS alonetreated group).
그림 Fig. 7. iNOS, COX-2 mRNA expression levels of hot water extracts from Mangifera indica L. peel in RAW 264.7 cells. RAW 264.7 cells (1 × 10⁶ cells/well) were pretreated with LPS (10 μg/ml) for 2 h and then treated water extracts from Mangifera indica L. peel at an appropriate concentration (10, 50, 100 and 500 μg/ml) for 16 h. iNOS and COX-2 protein expression was determined in RAW 264.7 cells treated with AMPW. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs LPS alone-treated group).
그림 Fig. 8. iNOS, COX-2 mRNA expression levels of 70% ethanol extracts from Mangifera indica L. peel in RAW 264.7 cells. RAW 264.7 cells (1 × 10⁶ cells/well) were pretreated with LPS (10 μg/ml) for 2 h and then treated ethanol extracts from Mangifera indica L. peel at an appropriate concentration (10, 50, 100 and 500 μg/ml) for 16 h. iNOS and COX-2 protein expression was determined in RAW 264.7 cells treated with AMPE. Results are means ± SD from triplicate experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs LPS alone-treated group).

참고문헌 (45)

Jung DH, Shin JH, Cho YY, Son JE. 2015. Development of a twovariable spatial leaf photosynthetic model of irwin mango grown in greenhouse. J. Bio-Env. Con. 24: 161-166.
An MR, Keum YS, Lee SK. 2015. Comparative analysis of volatile flavor compounds in Taiwan apple mango and Philippines Carabao Mango. Korean J. Food Sci. Technol. 47: 191-197.

원문보기 상세보기
Naef R, Velluz A, Jaquier A. 2006. The perfume of carabao mangoes (Mangifera indica L.): Identification of uncommon unsaturated fatty acid esters in the SPME of the intact fruit. Eur. Food Res. Technol. 222: 554-558.

상세보기
Ji ST, Youm JW, Yoo JY. 2018. A feasibility study on the cultivation of tropical fruit in Korea: Focused on Mango. JKAIS 19: 252-263.
Navarro M, Arnaez E, Moreira I, Quesada S, Azofeifa G, Wilhelm K, et al. 2019. Polyphenolic characterization, antioxidant, and cytotoxic activities of Mangifera indica cultivars from Costa Rica. Foods 8: 384.

상세보기
Lauricella M, Lo Galbo V, Cernigliaro C, Maggio A, Palumbo Piccionello A, Calvaruso G, et al. 2019. The anti-cancer effect of Mangifera indica L. peel extract is associated to γH2AX-mediated apoptosis in colon cancer cells. Antioxidants (Basel) 8: 422.
Lim K, Cabajar AA, Lobarbio C, Taboada EB, Lacks DJ. 2019. Extraction of bioactive compounds from mango (Mangifera indica L. var. Carabao) seed kernel with ethanol-water binary solvent systems. J. Food Sci. Technol. 56: 2536-2544.

상세보기
Alanon ME, Palomo I, Rodriguez L, Fuentes E, Arraez-Roman D, Segura-Carretero A. 2019. Antiplatelet activity of natural bioactive extracts from Mango (Mangifera Indica L.) and its by-products. Antioxidants (Basel) 8: 517.

상세보기
Dhananjaya BL, Shivalingaiah S. 2016. The anti-inflammatory activity of standard aqueous stem bark extract of Mangifera indica L. as evident in inhibition of Group IA sPLA2. An. Acad. Bras. Cienc. 88: 197-209.

상세보기
Kim MK. 2017. Comparison of antioxidant and antimicrobial activities of pulp and peel extracts of Mango. J. Invest. Cosmetol. 13: 113-118.

상세보기
Kim HK. 2018. The effects of fruit and vegetable bark extract on learning ability and memory improvement. JCCT 4: 261-267.
Jung ES. 2007. Antioxidant and antibacterial activity of extracts from Mangifera indica. Sunchon National University, Sunchun, Korea.
Sivakumar D, Jiang Y, Yahia EM. 2011. Maintaining mango (Mangifera indica L.) fruit quality during the export chain. Food Res. Int. 44: 1254-1263.

상세보기
Lee JY, Yoo DH, Joo DH, Kim SR, Jo HS, Joo SH, et al. 2017. Antiinflammatory effects of Amelanchier asiatica fruits ethanol extract. J. Soc. Cosmet. Scientists Korea, 43: 19-26.

원문보기 상세보기
Lee SH, Suh SJ, Lee KH, Yang JB, Choi SU, Park SS. 2013. Antiinflammatory effect of peel extracts from citrus fruits. J. Food Hyg. Saf. 28: 342-348.

원문보기 상세보기
Jeong DH, Kim KBWR, Kang BK, Jung SA, Kim HJ, Jeong HY, et al. 2012. Anti-inflammatory activity of the water extract of Sargassum fulvellum. KSBB J. 27: 325-329.

원문보기 상세보기
Jung YS, Eun CS, Jung YT, Kim HJ, Yu MH. 2013. Anti-inflammatory effects of Picrasma Quassioides (D.DON) BENN leaves extracts. J. Life Sci. 23: 629-636.

원문보기 상세보기
RodriguezVita J, Lawrence T. 2010. The resolution of inflammation and cancer. Cytokine Growth Fact. Rev. 21: 61-65.

상세보기
Lee JY, Yoo DH, Jeong YS, Joo SH, Chae JW. 2018. Verification of anti-inflammatory activities of the ethanol extracts of Glechoma hederacea var. longituba in RAW264.7 cells. J. Life Sci. 28: 429-434.

원문보기 상세보기
Roh KB, Lee JA, Park JH, Jung KS, Jung ES, Park DH. 2017. Antiinflammatory and anti-allergic effects of Gnaphalium affine extract. J. Soc. Cosmet. Sci. Korea 43: 103-114.

원문보기 상세보기
Knowles RG, Moncada S. 1994. Nitric oxide synthases in mammals. Biochem. J. 298: 249-258.

상세보기
Nathan C. 1997. Inducible nitric oxide synthase: what difference does it make?. J. Clin. Invest. 100: 2417-2423.

상세보기
Xue Q, Yan Y, Zhang R, Xiong H. 2018. Regulation of iNOS on immune cells and its role in diseases. Int. J. Mol. Sci. 19: 3805.
Kim HJ, Lee DJ, Ku JJ, Choi K, Park GW, Kang SH, et al. 2013. Antiinflammatory effect of extracts from folk plants in Ulleung Island. Korean J. Plant Res. 26: 169-177.

원문보기 상세보기
Kim JY, Jung KS, Jeong HG. 2004. Suppressive effects of the kahweol and cafestol on cyclooxygenase-2 expression in macrophages. FEBS Lett. 569: 321-326.

상세보기
Kwon YB, Yoo BS, Kim DS, Moon SJ, Yoon MS, Park SN. 2010. Anti inflammatory activity of Viburnum dilatatum Thunb. extract as cosmetic ingredient. J. Soc. Cosmet. Sci. Korea 36: 183-191.
Lee SH, Suh SJ, Lee KH, Yang JB, Choi SU, Park SS. 2013. AntiInflammatory effect of peel extracts from citrus fruits. J. Food Hyg. Saf. 28: 342-348.

원문보기 상세보기
Singleton VL, Orthofer R, Lamuela-Raventos RM. 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods Enzymol. 299: 152-178.
Blois MS. 1958. Antioxidant determinations by the use or a stable free radical. Nature 181: 1199-200.

상세보기
Roberta R, Nicoletta P, Anna P, Ananth P, Min Y, Catherine RE. 1999. Antioxidant acitivity applying an improved ABTS radical cation decolorization assay. Free Rad. Biol. Med. 26: 1231-1237.

상세보기
Jeong DH, Kim KBWR, Kang BK, Jung SA, Kim HJ, Jeong HY, et al. 2012. Anti-inflammatory activity of the water extract of Sargassum fulvellum. KSBB J. 27: 325-329.

원문보기 상세보기
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.

상세보기
Yoo YC, Lee GW, Cho YH. 2016. Antioxidant and anti-inflammatory effects of extracts from the flowers of Weigela subsessilis on RAW 264.7 macrophages. J. Life Sci. 26: 338-345.

원문보기 상세보기
Lee SO, Lee HJ, Yu MH, Im HG, Lee IS. 2005. Total polyphenol contents and antioxidant activities of methanol extracts from vegetables produced in Ullung Island. Korea J. Food Sci. Technol. 37: 233-240.
Jeong HJ, Park SB, Kim SA, Kim HK. 2007. Total polyphenol content and antioxidative activity of wild grape (Vitis coignetiae) extracts depending on ethanol concentrations. J. Korean Soc. Food Sci. Nutr. 36: 1491-1496.

원문보기 상세보기
Slater TF. 1984. Free-radical mechanisms in tissue injury. Biochem J. 222: 1-15.

상세보기
Yu MH, Im HG, Lee HJ, Ji YJ, Lee IS. 2006. Components and their antioxidative activities of methanol extracts from sarcocarp and seed of Zizyphus jujuba var. inermis Rehder. Korean J. Sci. Technol. 38: 128-134.
Jung SB. 2018. A study of the antioxidative activities of purple corn husk extracts as cosmetic ingredient. Silla University, Busan, Korea.
Jang DY. 2020. A Study on the effectiveness evaluation of Rhus javanica L. fruit extracts and the application of cosmetics. Mokwon University, Deajeon, Korea.
Jo HJ, Kim JW, Yoon JA, Kim KI, Chung KH, Song BC, et al. 2014. Antioxidant activities of Amaranth (Amaranthus spp. L.) flower extracts. Korean J. Food Nutr. 27: 175-182.

원문보기 상세보기
Kim YE, Yang JW, Lee CH, Kwon EK. 2009. ABTS radical scavenging and anti-tumor effects of Tricholoma matsutake sing. (Pine Mushroom). J. Korean Soc. Food Sci. Nutr. 38: 555-560.

원문보기 상세보기
Yoon WJ, Lee JA, Kim KN, Kim JY, Park SY. 2007. In vitro antiinflammatory activity of the Artemisia fukudo extracts in murine macrophage RAW264.7 cells. Korean J. Food Sci. Technol. 39: 464-469.
Cossenza M, Socodato R, Portugal CC, Domith IC, Gladulich LF, Encarnacao TG, et al. 2014. Nitric oxide in the nervous system: biochemical, developmental, and neurobiological aspects. Vitam. Horm. 96: 79-125.

상세보기
Golden BD, Abramson SB. 1999. Selective cyclooxygenase-2 inhibitor. Rheum. Dis. Clin. N. Am. 25: 359-378.

상세보기
Kang CH, Koo JR, So JS. 2012. Inhibitory Effects of Aralia cordata thunb extracts on nitric oxide synthesis in RAW264.7 macrophage cells. Korean J. Food Sci. Technol. 44: 621-627.

원문보기 상세보기

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].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증