[논문]이삭물수세미(Myriophyllum spicatum L.) 에탄올 추출물의 항산화와 항염증 효과

김철환; 이영경; 김민진; 최지수; 황병수; 조표연; 김영준; 정용태

doi:10.7732/kjpr.2023.36.1.015

이삭물수세미(Myriophyllum spicatum L.) 에탄올 추출물의 항산화와 항염증 효과
Evaluation Antioxidant and Anti-inflammatory Activity of Ethanolic Extracts of Myriophyllum spicatum L. in Lipopolysaccharide-stimulated RAW 264.7 Cells 원문보기

韓國資源植物學會誌 = Korean journal of plant resources, v.36 no.1, 2023년, pp.15 - 25

김철환 (국립낙동강생물자원관) , 이영경 (국립낙동강생물자원관) , 김민진 (국립낙동강생물자원관) , 최지수 (국립낙동강생물자원관) , 황병수 (국립낙동강생물자원관) , 조표연 (국립낙동강생물자원관) , 김영준 (고려대학교 식품규제과학과) , 정용태 (국립낙동강생물자원관)

초록
AI-Helper

이삭물수세미는 민간에서는 전초를 고름, 염증 등에 약용으로 사용하였으나, 염증에 대한 연구가 미비한 상황이다. 이에 본 연구에서는 이삭물수세미 추출물(EMS)의 항산화 효능과 항염증 효능을 분석하였다. 항산화 효능은 DPPH 라디칼 소거능과 환원력을 통해 산화적 스트레스를 통해 염증을 유발시킬 수 있는 ROS (Hong et al., 2020; Snezhkina et al., 2019)를 억제하는지 확인하였고, 항염증 효능은 염증 발현 인자인 LPS를 이용하여 RAW 264.7 대식세포에 염증을 유도한 뒤 pro-inflammatory cytokine (TNF-α, IL-1β)과 염증 매개체(NO, PGE₂)의 억제 및 TLR4/Myd88/NF-κB signaling pathway 발현 억제를 통해 확인하였다. 연구 결과, 항산화 효능에 있어서는 DPPH 라디칼 소거능과 Fe³⁺를 Fe²⁺로 환원시키는 환원력이 농도 의존적으로 증가함을 확인하였다. 무독성 상태에서 실험하기 위해 LPS와 EMS를 처리한 RAW 264.7 대식세포에서 90% 이상의 생존율을 나타내는 조건에서 실험을 진행하였다. LPS로 염증이 유도된 RAW 264.7 세포에서 EMS는 염증 매개 인자의 발현 및 생성 억제(iNOS에 의한 NO 생성 및 COX-2에 의한 PGE₂ 생성억제)와 pro-inflammatory cytokine (TNF-α 및 IL-1β)의 생성 또한 억제하였다. 특이적으로 COX-2에 의한 PGE₂ 생성 억제에서는 고농도에서 작용함을 확인하였고, IL-1β에서는 약한 억제력을 보였다. 이후 signaling pathway에서 염증 전사인자 경로를 확인하기 위하여 TLR4/MyD88의 활성을 확인하였고, EMS 처리에 따라 농도 의존적으로 억제되는 것을 확인하였다. 이에 따라 염증 초기 단계에서 NF-κB p65가 nuclear로 들어가는 것을 억제하는지 확인하기 위해 early time (LPS 처리 후 30, 60 min) 조건으로 nuclear에서 p65 인산화를 확인하였다. 그 결과, LPS 자극으로 인해 증가된 p65 인산화가 EMS에 의해 부분적으로 억제됨을 확인하였다. 이상의 결과를 통해 LPS로 염증이 유도된 RAW 264.7 대식세포에서 EMS가 COX-2에 의한 PGE₂ 생성 억제와 IL-1β의 생성에 있어 낮은 억제력을 가진 반면, iNOS에 의한 NO과 TNF-α 생성 및 TLR4/MyD88 singnaling pathway에 있어 강한 억제력을 가짐을 확인하였다. 결론적으로 EMS가 ROS를 제거하고 TLR4/MyD88/NF-κB signaling pathway를 억제함으로써 염증 인자들의 전사를 억제하고, 염증 인자 부분에서는 iNOS에 의한 NO 생성과 TNF-α 생성을 강하게 억제하여 RAW 264.7 대식세포에서 LPS로 자극된 염증을 억제하는 것으로 판단된다. 또한 TLR4/Myd88/NF-κB signaling pathway를 통한 pro-inflammatory cytokine과 염증 매개체와의 연관성에 대한 기초자료로 활용할 수 있는 근거 자료가 될 수 있을 것으로 생각된다.

Abstract ▼ AI-Helper

Myriophyllum spicatum L. has been used as an ornamental in ponds and aquariums, and as a folk remedy for inflammation and pus. Nevertheless, the biological activity and underlying mechanisms of anti-inflammatory effects are unclear. This study is aimed at investigating the antioxidative and anti-inflammatory activities of ethanol extract of Myriophyllum spicatum L. (EMS) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Antioxidant activity of EMS was assessed by radical-scavenging effects on ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals. As inflammatory response parameters produced by LPS-stimulated RAW 264.7 cells were quantified to assess the anti-inflammatory activity of EMS. Our results showed that EMS increased FRAP and DPPH radical-scavenging activity. In EMS-treated RAW 264.7 cells, the production of NO, PGE2, TNF-α and IL-1β was significantly inhibited at the non-cytotoxic concentration. In addition, EMS significantly attenuated LPS-stimulated the toll-like receptor (TLR) 4/myeloid differentiation protein (MyD) 88 signaling pathway, and inhibited nuclear translocation of nuclear factor-kappa B(NF-κB). Positive correlations were noted between anti-inflammatory activity and antioxidant activity. In conclusion, it was indicated that EMS suppresses the transcription of inflammatory factors by inhibiting the TLR4/MyD88/NF-κB signaling pathway, thereby suppressing LPS-stimulated inflammation in RAW 264.7 cells. This study highlights the potential role of EMS against inflammation and associated diseases.

주제어

표/그림 (7)

그림 Fig. 1. Antioxidant (DPPH, Reducing power) activities of ethanol extract of Myriophyllum spicatum L. (EMS). (A) DPPH free radical-scavenging activity and (B) Reducing power of EMS obtained with bioprotease in an in vitro cell-free experiments. Each value is presented as the mean ± SD and is representative of the results obtained from three independent experiments. The significance was determined by the Student's t-test (^*p < 0.05, compared with control).
그림 Fig. 2. Effect of EMS on the viability of RAW 264.7 cells. The cells were treated with the indicated concentrations of EMS alone (A) or in combination with lipopolysaccharide (LPS; 500 ng/mL) (B) for 24 h. Cell viability was assessed using the CCK-8 assay, and the results are expressed as the percentage of surviving cells over control cells (no addition of EMS). Each value is presented as the mean ± SD and is representative of the results obtained from three independent experiments. The significance was determined by the Student's t-test (^*p < 0.05, compared with control group; N.S., not significant).
그림 Fig. 3. Downregulation of nitric oxide (NO) and prostaglandin E₂ (PGE₂) production by EMS in LPS-stimulated RAW 264.7 cells. (A) Cells were pretreated with various concentrations of EMS (50, 100, 200, and 400 ㎍/mL) for 1 h prior to incubation with LPS (500 ng/mL) for 24 h, and nitrite content was measured using the Griess reaction. (B) The PGE₂concentration was measured in culture media using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Each value is presented as the mean ± SD and is representative of the results obtained from three independent experiments. The significance was determined by the Student's t-test (^*p < 0.05, compared with control group; ^#p < 0.05, compared with LPS).
그림 Fig. 4. Effect of EMS on LPS-stimulated iNOS and COX-2 expression in RAW 264.7 cells. RAW 264.7 cells were pretreated with EMS (50, 100, 200, and 400 ㎍/mL) 1 h prior to incubation with LPS (500 ng/mL) for 24 h. (A and B) Cell lysates were prepared for western blot analysis, with antibodies specific for murine iNOS and COX-2, and for an enhanced chemiluminescence (ECL) detection system. The experiment was repeated three times and similar results were obtained. β-actin was used as the internal controls for the western blot analysis, respectively.
그림 Fig. 5. Downregulation of tumor necrosis factor (TNF)-α and interleukin (IL)-1β production by EMS in LPS-stimulated RAW 264.7 cells. Cells were pretreated with various concentrations of EMS (50, 100, 200, and 400 ㎍/mL) for 1 h prior to incubation with LPS (500 ng/mL). After incubation for 24 h, the levels of TNF-α (A), IL-1β (B) present in the supernatants were measured using ELISA kits. Each value is presented as the mean ± SD and is representative of the results obtained from three independent experiments. The significance was determined by the Student's t-test (^*p < 0.05, compared with control group; ^#p < 0.05, compared with LPS).
그림 Fig. 6. Effect of EMS on LPS-stimulated TNF-α and IL-1β expression in RAW 264.7 cells. RAW 264.7 cells were pretreated with EMS (50, 100, 200, and 400 ㎍/mL) 1 h prior to incubation with LPS (500 ng/mL) for 24 h. (A and B) Cell lysates were prepared for western blot analysis, with antibodies specific for murine IL-1β and TNF-α, and for an ECL detection system. The experiment was repeated three times and similar results were obtained. β-actin was used as the internal controls for the western blot analysis, respectively.
그림 Fig. 7. Effects of EMS on LPS-stimulated TLR4 and NF-κB expression in RAW 264.7 cells. (A and B) RAW 264.7 macrophage cells were pretreated with EMS (50, 100, 200, and 400 ㎍/mL) 1 h prior to incubation with LPS (500 ng/mL) for 30 min. Cell lysates were prepared for western blot analysis, with antibodies specific for murine TLR4 and MyD88, and for an ECL detection system. (C and D) After 500 ng/mL LPS treatment with or without 400 μg/mL EMS for the indicated times(30, 60 min), the nuclear proteins were isolated, and the expression of NF-κB was determined by western blot analysis using an ECL detection system. The experiment was repeated three times and similar results were obtained. Nucleolin and β-actin were used as internal controls for the western blot analysis, respectively.

참고문헌 (49)

Anwar, M.A., S. Basith and S. Choi. 2013. Negative regulatory？approaches to the attenuation of toll-like receptor signaling.？Exp. Mol. Med. 45(2):e11.

상세보기
Atreya, R. and M.F. Neurath. 2005. Involvement of IL-6 in the？pathogenesis of inflammatory bowel disease and colon cancer.？Clin. Rex. Allergy Immunol. 28(3):187-196.
Babcock, T.A., W.S. Helton, K.N. Anwar, Y.Y. Zhao and N.J.？Espat. 2004. Synergistic anti-inflammatory activity of omega-3？lipid and rofecoxib pretreatment on macrophage proinflammatory cytokine production occurs via divergent NF-kappaB？activation. J. Parenter Enteral. Nutr. 28(4):232-239.
Braddock, M. and A. Quinn. 2004. Targeting IL-1 in inflammatory disease: New opportunities for therapeutic intervention.？Nat. Rev. Drug Discov. 3(4):330-339.

상세보기
Chainy, G.B.N. and D.K. Sahoo. 2020, Hormones and oxidative？stress: An overview. Free Radic. Res. 54(1):1-26.

상세보기
Cheel, J., C. Theoduloz, J. Rodriguez and G. Schmeda-Hirschmann.？2005. Free radical scavengers and antioxidants from lemongrass (Cymbopogon citratus (DC.) Stapf.). J. Agric. Food？Chem. 53(7):2511-2517.

상세보기
Chelombitko, M.A. 2018. Role of reactive oxygen species in？inflammation: A minireview. Moscow Univ. Biol. Sci. Bull.？73(4):199-202.

상세보기
Cheng, B.C.Y., X.Q. Ma, H.Y. Kwan, K.W. Tse, H.H. Cao, T.？Su, X. Shu, Z.Z. Wu and Z.L. Yu. 2014. A herbal formula？consisting of Rosae Multiflorae Fructus and Lonicerae？Japonicae Flos inhibits inflammatory mediators in LPS-stimulated RAW 264.7 macrophages. J. Ethnopharmacol.？153(3):922-927.

상세보기
Choi, H.Y., J.E. Lim and J.H. Hong. 2010. Curcumin interrupts？the interaction between the androgen receptor and Wnt/β？-catenin signaling pathway in LNCaP prostate cancer cells.？Prostate Cancer Prostatic Dis. 13(4):343-349.

상세보기
Church, L.D. and M.F. McDermott. 2009. Canakinumab, a？fully-human mAb against IL-1beta for the potential treatment？of inflammatory disorders. Curr. Opin. Mol. Ther. 11(1):81-89.

상세보기
Coleman, J.W. 2001. Nitric oxide in immunity and inflammation.？Int. Immunopharmacol. 1(8):1397-1406.

상세보기
Cui, E.J., H.N. Lyu, M.C. Song, H.J. Yang, S.J. Nam, S.K. Park,？H.K. Choi, I.S. Chung and N.I. Baek. 2008. Isolation and？identification of Quercitrin from Myriophyllum spicatum L.？Korean J. Medicinal Crop Sci. 27:62-65.
Dinarello, C.A. 1996. Biological basis for Interleukin-1 in？disease. Blood 87(6):2095-2147.

상세보기
Dinarello, C.A. 2004. Therapeutic strategies to reduce IL-1？activity in treating local and systemic inflammation. Curr.？Opin. Pharmacol. 4:378-385.

상세보기
Fang, H., P.F. Wang, Y. Zhou, Y.C. Wang and Q.W. Yang. 2013.？Toll-like receptor 4 signaling in intracerebral hemorrhage-induced inflammation and injury. J. Neuroinflammation？10(1):1-10.
Feghali, C.A. and T.M. Wright. 1997. Cytokines in acute and？chronic inflammation. Front. Biosci. 2(4):12-26.
Fujihara, M., M. Muroi, K. Tanamoto, T. Suzuki, H. Azuma？and H. Ikeda. 2003. Molecular mechanisms of macrophage？activation and deactivation by lipopolysaccharide: Roles of？the receptor complex. Pharmacol. Ther. 100:171-194.

상세보기
Gross, E.M., H. Meyer and G. Schilling. 1996. Release and？ecological impact of algicidal hydrolysable polyphenols in？Myriophyllum spicatum. Phytochemistry 41(1):133-138.

상세보기
Guha, M. and N. Mackman. 2001. LPS induction of gene？expression in human monocytes. Cell. Signal. 13(2):85-94.

상세보기
Guzik, T., R. Korbut and T. Adamek-Guzik. 2003. Nitric oxide？and superoxide in inflammation. J. Physiol. Pharmacol. 54(4):469-487.
Hong, S., P. Pangloli, R. Perumal, S. Cox, L.E. Noronha, V.P.？Dia and D. Smolensky. 2020. A comparative study on phenolic content, antioxidant activity and anti-inflammatory？capacity of aqueous and ethanolic extracts of sorghum in？lipopolysaccharide-induced RAW 264.7 macrophages. Antioxidants 9(12):1297.

상세보기
Jeong, J.W., H.H. Lee, M.H. Han, G.Y. Kim, W.J. Kim and？Y.H. Choi. 2014. Anti-inflammatory effects of genistein via？suppression of the toll-like receptor 4-mediated signaling？pathway in lipopolysaccharide-stimulated BV2 microglia.？Chem. Biol. Interact. 212:30-39.

상세보기
Jones, N, J. Blagih, F. Zani, A. Rees, D.G. Hill, B.J. Jenkins, C.J.？Bull, D. Moreira, A.I.M. Bantan, J.G. Cronin, D. Avancini,？G.W. Jones, D.K. Finlay, K.H. Vousden, E.E. Vincent and？C.A. Thornton. 2021. Fructose reprogrammes glutamine-dependent oxidative metabolism to support LPS-induced？inflammation. Nat. Commun. 12(1):1209.

상세보기
Kawahara, K., H. Hohjoh, T. Inazumi, S. Tsuchiya and Y. Sugimoto. 2015. Prostaglandin E2-induced inflammation: Relevance of prostaglandin E receptors. Biochim. Biophys. Acta.？1851(4):414-421.
Kim, C.H., Y.K. Lee, J.W. Jeong, B.S. Hwang, Y.T. Jeong,？Y.T. Oh, P.Y. Cho and C.H. Kang. 2021a. Anti-inflammatory？effects of Hemistepta lyrata Bunge in LPS-stimulated RAW？264.7 cells through regulation of MAPK signaling pathway.？Korean J. Plant Res. 34(1):23-30.

원문보기 상세보기
Kim, D.H., E.Y. Hwang and J.H. Son. 2013. Anti-inflammatory？activity of Carthamus tinctorious seed extracts in Raw 264.7？cells. J. Life Sci. 23(1):55-62 (in Korean).

원문보기 상세보기
Kim, D.S., H.J. Eo, Y. Kang and G.H. Park. 2021b. Anti-inflammatory effect of Berchemia berchemiaefolia leaves？through inhibition of NF-κB and MAPK signaling activation？in LPS-stimulated RAW264.7 cells. Korean J. Plant Res.？34(1):31-36.

원문보기 상세보기
Kim, J.H., H.N. Song, H.C. Ko, J.Y. Lee, M.G. Jang and S.J.？Kim. 2017. Anti-oxidant and anti-inflammatory properties？of Clerodendrum trichotomum leaf extracts. J. Life Sci. 27(6):640-645 (in Korean).

원문보기 상세보기
Kim, S.H. and S.A. Kang. 2019. Anti-inflammatory effects of？beopje processed curly dock (Rumex crispus L.) in LPS-induced murine RAW 264.7 cell lines. Korean J. Food &？Nutr. 32(5):408-416 (in Korean).
Laksmitawati, D.R., A.P. Prasanti, N. Larasinta, G.A. Syauta,？R. Hilda, H.U. Ramadaniati, A. Widyastuti, N. Karami, M.？Afni, D.D. Rihibiha, W. Widowati and H.S.W. Kusuma.？2016. Anti-inflammatory potential of gandarusa (Gendarussa？vulgaris Nees) and soursoup (Annona muricata L) extracts？in LPS stimulated-macrophage cell (RAW264.7). J. Nat.？Remedies 16(2):73-81.
Lee, S.H., E.J. Lee, C. Chung, H.Y. Sohn and J.S. Kim. 2019.？Hesperetin ameliorates inflammatory responses in lipopolysaccharide-stimulated RAW 264.7 cells via p38 MAPK and？ERK1/2. J. Life Sci. 29(1):129-134 (in Korean).

원문보기 상세보기
Lee, W.J. 2011. Fiber type specific expression of toll-like？receptor4, IL-6, TNF-α, and suppressor of cytokine signaling-3 after acute exercise in rat skeletal muscles. J. Life Sci.？21(9):1259-1265 (in Korean).

원문보기 상세보기
Li, G.Z., Y. Zhang, J.B. Zhao, G.J. Wu, X.F. Su and C.H. Hang.？2011. Expression of myeloid differentiation primary response？protein 88 (Myd88) in the cerebral cortex after experimental？traumatic brain injury in rats. Brain Res. 1396:96-104.

상세보기
Lopez-Castejon, G. and D. Brough. 2011. Understanding the？mechanism of IL-1β secretion. Cytokine Growth Factor？Rev. 22(4):189-195.
Melendez-Martinez, A.J. 2019. An overview of carotenoids,？apocarotenoids, and vitamin A in agro-food, nutrition, health,？and disease. Mol. Nutr. Food Res. 63:e1801045.

상세보기
Nakai, S., S. Yamada and M. Hosomi. 2005. Anti-cyanobacterial？fatty acids released from Myriophyllum spicatum. Hydrobiologia 543(1):71-78.

상세보기
Nakai, S., Y. Inoue, M. Hosomi and A. Murakami. 2000. Myriophyllum spicatum-released allelopathic polyphenols inhibiting？growth of blue-green algae Microcystis aeruginosa. Water？Res. 34(11):3026-3032.

상세보기
Needleman, P. and P.C. Isakson. 1997. The discovery and？function of COX-2. J. Rheumatol. Suppl. 49:6-8.
Nemmar, A, S. Beegam, N.E. Zaaba, S. Alblooshi, S. Alseiari？and B.H. Ali. 2022. The salutary effects of catalpol on diesel？exhaust particles-induced thrombogenic changes and cardiac？oxidative stress, inflammation and apoptosis. Biomedicines？10(1):99.

상세보기
Parameswaran, N. and S. Patial. 2010. Tumor necrosis factor-α？signaling in macrophages. Crit. Rev. Eukaryot. Gene Expr.？20(2):87-103.

상세보기
Ren, K. and R. Torres. 2009. Role of interleukin-1β during pain？and inflammation. Brain Res. Rev. 60(1):57-64.

상세보기
Sambon, M, A. Gorlova, A. Demelenne, J. Alhama-Riba, B.？Coumans, B. Lakaye, P. Wins, M. Fillet, D.C. Anthony, T.？Strekalova and L. Bettendorff. 2020. Dibenzoylthiamine has？powerful antioxidant and anti-inflammatory properties in？cultured cells and in mouse models of stress and neurodegeneration. Biomedicines 8(9):361.

상세보기
Seo, H.J. and J.B. Jeong. 2020. Immune-enhancing effects of？green lettuce (Lactuca sativa L.) extracts through the TLR4-MAPK/NF-κB signaling pathways in RAW264.7 macrophage cells. Korean J. Plant Res. 33(3):183-193.

원문보기 상세보기
Seo, K.H., J.Y. Park, H.J. Noh, J.Y. Lee, E.Y. Lee, J.G. Han,？J.H. Kim and M.S. Cheong. 2018. Anti-inflammatory effects？of various mushrooms in LPS-stimulated RAW264.7 cells.？Korean J. Plant Res. 31(5):478-488.

원문보기 상세보기
Snezhkina, A.V., A.V. Kudryavtseva, O.L. Kardymon, M.V.？Savvateeva, N.V. Melnikova, G.S. Krasnov and A.A. Dmitriev. 2019. ROS generation and antioxidant defense systems？in normal and malignant cells. Oxid. Med. Cell. Longev. 2019:6175804.

상세보기
Westman, M., M. Korotkova, E.A. Klint, A. Stark, L.P. Audoly,？L. Klareskog, A.K. Ulfgren and P.J. Jakobsson. 2004. Expression of microsomal prostaglandin E synthase 1 in rheumatoid？arthritis synovium. Arthritis Rheum. 50(6):1774-1780.
Yen, G.C. and H.Y. Chen. 1995. Antioxidant activity of various？tea extracts in relation to their antimutagenicity. J. Agric.？Food Chem. 43:27-32.

상세보기
Yoon, Y.I., J.S. Hwang, M.Y. Ahn, Y.B. Lee, M.S. Han, T.W.？Goo and E.Y. Yun. 2015. Inhibition of inflammation by？Popillia flavosellata ethanol extract in LPS- induced RAW？264.7 macrophages. J. Life Sci. 25(9):993-999.

원문보기 상세보기
Zhu, H.T., C. Bian, J.C. Yuan, W.H. Chu, X. Xiang, F. Chen,？C.S. Wang, H. Feng and J.K. Lin. 2014. Curcumin attenuates？acute inflammatory injury by inhibiting the TLR4/MyD88/NF-κB signaling pathway in experimental traumatic brain？injury. J. Neuroinflammation. 11(1):1-17.

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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