[논문]포공영, 금은화, 익모초 혼합물의 항산화 및 항염증 효과에 관한 연구

허성신; 김영일

doi:10.13048/jkm.23027

포공영, 금은화, 익모초 혼합물의 항산화 및 항염증 효과에 관한 연구
The Study on Antioxidant and Anti-inflammatory Effects of Taraxacum platycarpum H. Dahlstedt, Lonicera japonica Thunberg and Leonurus japonicus Houtt. Complex 원문보기

Journal of Korean Medicine = 대한한의학회지, v.44 no.3, 2023년, pp.10 - 28

허성신 (대전대학교 한의과대학 침구학교실) , 김영일 (대전대학교 한의과대학 침구학교실)

Abstract ▼ AI-Helper

Objectives: This study was designed to experiment with the antioxidant and anti-inflammatory effects of Taraxacum platycarpum H. Dahlstedt, Lonicera japonica Thunberg, and Leonurus japonicus Houtt. complex (TLL) in LPS-induced RAW264.7 cell. Methods: The antioxidant activity of TLL was measured by FRAP assay, DPPH radical scavenging activity, ABTS radical scavenging activity. Total polyphenol and flavonoid contents of TLL were measured by using standard methods. The anti-inflammatory effects of TLL were measured by NO production, biomarker production (PGE2, IL-1β, IL-6, TNF-α), mRNA expression level (iNOS, COX-2, IL-1β, IL-6, TNF-α) and protein expression level (ERK, JNK, p38). Results: Total polyphenol and flavonoid contents in TLL were 58.03±1.02 mg of Gallic acid equivalents (GAE)/g and 16.58±0.60 mg of Quercetin equivalents (QE)/g respectively. In FRAP assay, DPPH and ABTS radical scavenging activity, a concentration-dependent increase in TLL was observed. To explore antioxidant and anti-inflammatory effects of TLL, RAW 264.7 cells were treated with TLL and LPS for 24 hours. Cell viability of RAW 264.7 cells were measured by adding EZ-Cytox, It was remarkably increased at 50, 100, 200 ㎍/㎖ concentrations of TLL. NO, ROS, iNOS, IL-1β, IL-6, TNF-α, ERK, JNK and p38 were remarkably decreased at 50, 100, 200 ㎍/㎖ concentrations of TLL compared to the control group. PGE2 and COX-2 were remarkably decreased at 100, 200 ㎍/㎖ concentrations Conclusion: These results suggest that TLL complex has antioxidant and anti-inflammatory effects.

주제어

표/그림 (19)

표 Table 1. The Sequence of Primers in Real-Time PCR
그림 Fig. 1. DPPH Radical Scavenging Activity of TLL Each concentration (1, 10, 100, and 1000 ㎍/㎖) of TLL was incubated with DPPH solution for 30 min. Activities were determined by measuring absorbance at 517 ㎚. The results were provided as mean ± SEM (n=3)
그림 Fig. 3. FRAP Value of TLL. Each concentration (1, 5, 10, and 50 ㎍/㎖) of TLL was incubated with FRAP regent solution for 30 min. Activities were determined by measuring absorbance at 593 ㎚. The results were provided as mean ± SEM (n=3)
그림 Fig. 2. ABTS Radical Scavenging Activity of TLL Each concentration (1, 10, 100, and 1000 ㎍/㎖) of TLL was incubated with ABTS solution for 10 min. Activities were determined by measuring absorbance at 732 ㎚. The results were provided as mean ± SEM (n=3)
그림 Fig. 4. Cell viability of TLL in RAW264.7 cell. RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL for 24 h. EZ-Cytox was added to treated cells and reacted for 30 min, thereafter microplate reader was used for measuring absorbance of the solution at 450 ㎚. Cell viability was calculated as percentage with regard to the control. The results were provided as mean ± SEM (n=3)
그림 Fig. 5. Effect of TLL on ROS level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100 and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. ROS levels were calculated as percentage with regard to the control. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, *: p<0.05, **: p<0.01, ***: p <0.001 compared to control).
그림 Fig. 6. Effect of TLL on NO level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100 and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. NO levels were calculated as percentage with regard to the control. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, ***: p<0.001 compared to control).
그림 Fig. 7. Effect of TLL on PGE2 level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. Treated cells were measured by mouse PGE₂ ELISA kit. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, ***: p<0.001 compared to control).
그림 Fig. 9. Effect of TLL on IL-6 level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. Treated cells were measured by mouse IL-6 ELISA kit. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, **: p<0.01, ***: p<0.001 compared to control).
그림 Fig. 8. Effect of TLL on IL-1β level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. Treated cells were measured by mouse IL-1beta ELISA kit. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, **: p<0.01, ***: p<0.001 compared to control).
그림 Fig. 10. Effect of TLL on TNF-α level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. Treated cells were measured by mouse TNF-α ELISA kit. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, *: p<0.05, ***: p<0.001 compared to control).
그림 Fig. 11. Effect of TLL on iNOS mRNA expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖ of TLL with 100 ng/㎖) LPS for 24 h. The mRNA levels of iNOS expression were measured by polymerase chain reaction. The normal group was not treated LPS. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, *: p<0.05, **: p<0.01, ***: p<0.001 compared to control).
그림 Fig. 13. Effect of TLL on IL-1β mRNA expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The mRNA levels of IL-1β expression were measured by polymerase chain reaction. The normal group was not treated LPS. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, **: p<0.01, ***: p<0.001 compared to control).
그림 Fig. 12. Effect of TLL on COX-2 mRNA expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The mRNA levels of COX-2 expression were measured by polymerase chain reaction. The normal group was not treated LPS. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, ***: p<0.001 compared to control).
그림 Fig. 14. Effect of TLL on IL-6 mRNA expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The mRNA levels of IL-6 expression were measured by polymerase chain reaction. The normal group was not treated LPS. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, **: p<0.01, ***: p<0.001 compared to control).
그림 Fig. 15. Effect of TLL on TNF-α mRNA expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The mRNA levels of TNF-α expression were measured by polymerase chain reaction. The normal group was not treated LPS. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p <0.001 compared to normal, *: p<0.05, ***: p<0.001 compared to control).
그림 Fig. 16. Effect of TLL on ERK protein expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The total cell extracts were subjected to western blot and 10% SDS-PAGE analysis with the respective primary and secondary antibodies. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, **: p<0.01, ***: p <0.001 compared to control).
그림 Fig. 17. Effect of TLL on JNK protein expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The total cell extracts were subjected to western blot and 10% SDS-PAGE analysis with the respective primary and secondary antibodies. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, ***: p<0.001 compared to control).
그림 Fig. 18. Effect of TLL on p38 protein expression level in RAW264.7 cell RAW264.7 cells were treated by each concentration (50, 100, and 200 ㎍/㎖) of TLL with 100 ng/㎖ LPS for 24 h. The total cell extracts were subjected to western blot and 10% SDS-PAGE analysis with the respective primary and secondary antibodies. The results were provided as mean ± SEM (n=3) (Significance of results, +++: p<0.001 compared to normal, ***: p<0.001 compared to control).

참고문헌 (41)

Reece JB, Urry LA, Cain ML, Wasserman SA,？Minorsky PV, Jackson RB, & Jeon SH tr.？Campbell Biology (10th ed). Seoul: Bio？Science. 2016: 202-6.
Pack JH, & Kweon GR. (2013). Clinical？Applications of Antioxidants. Hanyang Medical？Reviews. 33(2), 130-6. http://dx.doi.org/10.7599/hmr.2013.33.2.130

상세보기
Coyle JT, & Puttfarcken P. (1993). Oxidative？stress, glutamate, and neurodegenerative disorders.？Science. 262(5134), 689-95. doi: 10.1126/science.7901908

상세보기
Kwag SG, Kwon SY, Kim SK, Min BW, Seo？YM, Lee JM, et al. Basic Pathology. Seoul:？Jungmunkag. 2010: 84.
Punt J, Stranford SA, Jones PP, Owen JA, &？The Korean Society for Microbiology tr. Kuby？Immunology (8th ed). Seoul: Panmunedu. 2020:？142-3.
Abbas AK. Lichtman AH, Pillai S, & Kim PH？tr. Basic Immunology: Functions and Disoders？of the Immune System (6th ed). Seoul:？Panmunedu. 2020: 35, 44, 47.
Park MG, Yoo JD, & Lee KH. (2020). Current？Guidelines for Non-Steroidal Anti-Inflammatory？Drugs. J Korean Orthop Assoc. 55(1), 9-28.？https://doi.org/10.4055/jkoa.2020.55.1.9

상세보기
Golan DE, Armstrong EJ, Armstrong AW, &？Kim IK tr. Principles of Pharmacology: The？pathophysiologic Basis of Drug Therapy (4th？ed). Seoul: Shinilbooks. 2017: 795-8, 811.
Cha HY, Jeong AR, Cheon JH, Ahn SH, Park SY, & Kim KB. (2015). The Anti-oxidative？and Anti-inflammatory Effect of Lonicera？Japonica on Ulcerative Colitis Induced by？Dextran Sulfate Sodium in Mice. Korean J？Pediatr. 29(3), 54-64. http://dx.doi.org/10.7778/jpkm.2015.29.3.054

원문보기 상세보기
Choi YN, Choi YK, Nan L, & Choo BK.？(2020). Anti-oxidant and Anti-inflammatory？Effects of Ethanol Extracts from Leonurus？japonicus Houtt. on LPS-induced RAW 264.7？Cells. Korean J Org Agri. 28(4), 659-77.？http://dx.doi.org/10.11625/KJOA.2020.28.4.659

원문보기 상세보기
Lee YS. (2020). Anti-aging, Antioxidant and？Anti-inflammation Activities of Lonicerae？Flos, Dictamni Radicis Cortex and Forsythiae？Fructus Extracts. Journal of Investigative？Cosmetology. 16(1), 1-10. https://doi.org/10.15810/jic.2020.16.1.001
Herbology Editorial Committee of Korean？Medicine schools. Herbology [Boncho-hak]？(4th ed). Seoul: Young-Lim Press. 2020:？236-8, 240-2, 452-3.
Noh KH, & Baek JH. (2009). Inhibitory Effect？of Taraxaci Herba Extract (THE) on？Pro-inflammatory Mediatory. The Journal of？Korean Oriental Pediatrics. 23(3), 165-76.
Kang JR. (2009). Anti-oxidation and Melanin？Inhibition Characteristics of Taraxacum？platycarpum and Compositae plants. Ph.D.？thesis, Konkuk University.
So MS. (2009). Effect of Taraxacum mongolieum？Hand-Mass on Proliferation of Breast Cancer？Cells and Nitric Oxide Production. Journal of？Korean Health & Fundamental Medical Science.？2(3), 128-31.
Jo JH, Lyu JH, Kim CH, Kang KH, Yoon HJ,？Lee SY, et al. (2007). The Anti-allergic Effects？of Taraxaci Herba on the RBL-2H3 Cells. J？Korean Med Ophthalmol Otolaryngol Dermatol.？20(1), 209-17.
Lee HW, & Ma CJ. (2021). Neuroprotective？Activity of Lonicerin Isolated from Lonicera？japonica. Kor J Pharmacogn. 52(1), 19-25.？https://doi.org/10.22889/KJP.2021.52.1.19

원문보기 상세보기
Ahn SH, & Kim HH. (2010). Lonicerae Flos？Inhibited COX-2 and MMP-9 in LPS Induced？Arthritis of Mouse through Regulation of MIF.？Korean J Oriental Physiology & Pathology.？24(2), 242-8.
Park CW. Herbal Pharmacology [Hanyak-yakri-hak]. Seoul: Shin Kwang Pub. 2005:？169, 243-4.
Nho JH, Lee HJ, Jang JH, Yang BD, Woo？KW, Kim HA, et al. (2019). Protective Effect？of Water Extract of Leonurus japonicus？Houttuyn against HCl/EtOH-induced Gastric？Mucosal Damage and Genotoxicity Evaluation？using Micronucleus Test. Korean J. Plant Res.？32(4), 282-9. https://doi.org/10.7732/kjpr.2019.32.4.282

원문보기 상세보기
Xie M. Formulas of Chinese Medicine [fangji-xue].？Beijing: People's Medical Pubishing House.？2008: 453-4.
Park JH, Park JY, & Park SD. (2014).？Anti-Oxidative and Anti-Inflammatory Effect？of 7 Herbal Extracts and Methods of Herbal？Formula Compositioning. Herbal Formula？Science. 22(2): 87-103. doi: 10.14374/HFS.2014.22.2.087

원문보기 상세보기
Hong SJ, Choi EG, Lim HS, Shon JB, & Jeong？SS. (2001). Effect of herbal dentifrice on？dental plaque and gingivitis. Korean Academy of Dental Health. 25(4): 347-355.
Choi KM, Jeon JH, Kim ES, Sung KJ, & Kim？YI. (2021). Study on Inflammatory-Control？Mechanisms of Lonicera japonica Thunberg,？Forsythia viridissima Lindley, and Taraxacum？platycarpum H. Dahlstedt Complex. Journal of？Haehwa Medicine, 30(1): 11-31.
Lee SB. (2021). Anti-inflammatory Effects of？Taraxacum platycarpum H. Dahlstedt, Lonicera？japonica Thunberg, and Houttuynia cordata？Thunberg Complex in Lipopolysaccharide-Stimulated Macrophage and Mice. M.S.？thesis, Daejeon University.
Moon SC. (2022). Antioxidant and Anti-inflammatory Effects of Taraxacum？platycarpum H. Dahlstedt, Lonicera japonica？Thunberg, Citrus unshiu Markovich complex.？M.S. thesis, Daejeon University.
Zorov DB, Juhaszova M, & Sollott SJ. (2014).？Mitochondrial reactive oxygen species (ROS)？and ROS-induced ROS release. Physiol Rev.？94(3), 909-50. doi: 10.1152/physrev.00026.2013

상세보기
Li R, Jia Z, & Trush MA. (2016). Defining？ROS in Biology and Medicine. Reactive Oxygen？Species (Apex). 1(1), 9-21. doi: 10.20455/ros.2016.803
Frankel EN. (1996). Antioxidants in lipid foods？and their impact on quality. Food Chemistry.？57(1), 51-5. https://doi.org/10.1016/0308-8146(96)00067-2

상세보기
Kim HC. Herbal Pharmacology [Hanyak-yakri-hak]. Seoul: Jipmoon. 2001: 150-2, 155-6,？334-6.
Yang JH, Yoo HR, Kim YS, & Seol IC.？(2021). The Effect of Lonicera Japonica？Thunberg on Inflammatory Factor Expression？Associated with Atherosclerosis. J Int Korean？Med. 42(1), 25-39. https://doi.org/10.22246/jikm.2021.42.1.25

원문보기 상세보기
Kim TY, Jang SA, Chae YB, & Bak JP.？(2016). Antioxidant and Protective Effects of？Leonurus sibiricus L. Extract on Ultraviolet B？(UVB)-induced Damage in Human Keratinocytes.？Korean J. Plant Res. 29(1), 11-9. http://dx.doi.org/10.7732/kjpr.2016.29.1.011

원문보기 상세보기
Yoon HJ, Moon ME, Park HS, Im SY, & Kim？YH. (2007) Chitosan oligosaccharide (COS)？inhibits LPS-induced inflammatory effects in？RAW 264.7 macrophage cells. Biochemical？and Biophysical Research Communications.？358(3), 954-9. doi: 10.1016/j.bbrc.2007.05.042

상세보기
Shin JH, & Yoo SK. (2012). Antioxidant？Properties in Microbial Fermentation Products？of Lonicera japonica Thunb. Extract. Journal？of the East Asian Society of Dietary Life.？22(1), 95-102.
Ozben T. (2007). Oxidative stress and apoptosis:？impact on cancer therapy. Journal of Pharmaceutical？Sciences. 96(9), 2181-96.
Moloney JN, & Cotter TG. (2018). ROS？signalling in the biology of cancer. Semin Cell？Dev Biol. 80, 50-64. doi: 10.1016/j.semcdb.2017.05.023

상세보기
Yim CY. (2010). Nitric oxide and cancer.？Korean J Med. 78(4), 430-6.
Jin JR, Kim JH, Lim JH, Jung YH, & Kim？KH. PP+ Immunology. Seoul: Life Science.？2020: 40.
Kim MJ, Park SH, Kim KBWR, Choi JS, &？Ahn DH. (2017). Anti-Inflammatory Effect of？Chondria crassicaulis Ethanol Extract on？MAPKs and NF-κB Signaling Pathway in LPS-Induced RAW 264.7 Macrophages. KSBB？Journal. 32(4), 352-60. http://dx.doi.org/10.7841/ksbbj.2017.32.4.352

상세보기
Chan ED, & Riches DW. (2001). IFN-gamma？+ LPS induction of iNOS is modulated by？ERK, JNK/SAPK, and p38(mapk) in a mouse？macrophage cell line. Am J Physiol Cell？Physiol. 280(3), C441-50. https://doi.org/10.1152/ajpcell.2001.280.3.C441

상세보기
Murakami-Mori K, Mori S, & Nakamura S.？(1999). p38MAP kinase is a negative regulator？for ERK1/2-mediated growth of AIDS-associated？Kaposi's sarcoma cells. Biochem Biophys Res？Commun. 264(3), 676-82. https://doi.org/10.1006/bbrc.1999.1574

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저자의 다른 논문 :

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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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