The Korea Journal of Herbology (대한본초학회지)

The Korea Association of Herbology (대한본초학회)
권호 표지
DOI QR코드

DOI QR Code

Anti-inflammatory Effects of Epimedii Herba Water Extract through Inhibition of Nuclear Factor-κB in RAW 264.7 Cells

RAW 264.7 세포에서 음양곽(淫羊藿) 물 추출물의 nuclear factor-κB 억제를 통한 항염증 효과

  • Jung, Ji Yun (College of Korean Medicine, Daegu Haany University) ;
  • Byun, Sung Hui (College of Korean Medicine, Daegu Haany University) ;
  • Park, Chung A (College of Korean Medicine, Daegu Haany University) ;
  • Cho, Il Je (College of Korean Medicine, Daegu Haany University) ;
  • Kim, Sang Chan (College of Korean Medicine, Daegu Haany University)
  • 정지윤 (대구한의대학교 한의과대학) ;
  • 변성희 (대구한의대학교 한의과대학) ;
  • 박정아 (대구한의대학교 한의과대학) ;
  • 조일제 (대구한의대학교 한의과대학) ;
  • 김상찬 (대구한의대학교 한의과대학)
  • Received : 2017.12.22
  • Accepted : 2018.03.15
  • Published : 2018.03.30
Download PDF
⟨ Previous Next ⟩

Abstract

Objectives : Epimedii Herba has been frequently used in Korean Traditional Medicine to treat impotence, spermatorrhoea, exophthalmos, and forgetfulness. Present study investigated anti-inflammatory effects of Epimedii Herba water extract (EWE) and attempted to elucidate molecular mechanisms involved. Methods : To explore anti-inflammatory effects of EWE, RAW 264.7 cells, a murine macrophage cell line, were pretreated with $10-100{\mu}g/m{\ell}$ of EWE, and then subsequently exposed to $1{\mu}g/m{\ell}$ of lipopolysaccharide (LPS). Levels of nitric oxide (NO), interleukin-6, $interleukin-1{\beta}$, and tumor necrosis $factor-{\alpha}$ were monitored in the medium. Expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by immunoblot and real-time PCR analyses. Signaling pathways related with nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinases were monitored to elucidate molecular mechanisms involved. Finally, the role of three flavonoid compounds in EWE on LPS-mediated NO production were investigated. Results : In conditioned medium, pretreatment of EWE ($100{\mu}g/m{\ell}$) significantly inhibited LPS-stimulated NO and pro-inflammatory cytokine production. In addition, EWE attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by LPS. EWE prevented the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}$, nuclear translocation of $NF-{\kappa}B$, and DNA binding of $NF-{\kappa}B$, while EWE did not change the phosphorylation of mitogen-activated protein kinases by LPS. Moreover, icariin, icaritin, and quercetin partly, but significantly, inhibited the LPS-stimulated NO production. Conclusions : These results suggest that EWE has an ability to prevent inflammation in macrophages through inhibition of $NF-{\kappa}B$ signaling pathway.

Keywords

References

  1. Tacke F. Targeting hepatic macrophages to treat liver diseases. J Hepatol. 2017 ; 66 : 1300-12. https://doi.org/10.1016/j.jhep.2017.02.026
  2. Abbas AK, Lichtman AH, Phillai S. Cellular and molecular immunology. 7th ed. Philadelphia : Saunders. 2012 : 41-88.
  3. Kuzmich NN, Sivak KV, Chubarev VN, Porozov YB, Savateeva-Lyubimova TN, Peri F. TLR4 signaling pathway modulators as potential therapeutics in inflammation and sepsis. Vaccines. 2017 ; 5 : 34. https://doi.org/10.3390/vaccines5040034
  4. Lee SI. Herbology. Seoul : Suseowon. 1981 : 71-2.
  5. Kang BS, Ko UC, Kim SH, Rho SH, Song HJ, Shin MK, Ahn DK, Lee SI, Lee YJ, Lee TH, Ju YS. Herbology. Seoul : Younglimsa. 1992 : 553-4.
  6. Park HJ, Koo YK, Park MJ, Hwang YK, Hwang SY, Park NC. Restoration of spermatogenesis using a new combined herbal formula of Epimedium koreanum Nakai and Angelica gigas Nakai in an luteinizing hormone-releasing hormone agonist-induced ratmodel of male infertility. World J Mens Health. 2018, in press.
  7. Wu L, Du ZR, Xu AL, Yan Z, Xiao HH, Wong MS, Yao XS, Chen WF. Neuroprotective effects of total flavonoid fraction of the Epimedium koreanum Nakaiextract on dopaminergic neurons: In vivo and in vitro. Biomed Pharmacother. 2017 ; 91 : 656-63. https://doi.org/10.1016/j.biopha.2017.04.083
  8. Luo G, Gu F, Zhang Y, Liu T, Guo P, Huang Y. Icariside II promotes osteogenic differentiation of bone marrow stromal cells in beagle canine. Int J Clin Exp Pathol. 2015 ; 8 : 4367-77.
  9. Jung JY, Park SM, Ko HL, Lee JR, Park CA, Byun SH, Ku SK, Cho IJ, Kim SC. Epimedium koreanum Nakai water extract ameliorates oxidative stressmediated liver injury by activating nuclear factor erythroid 2-related factor 2. Am J Chin Med. 2018; 46 : 1-20. https://doi.org/10.1142/S0192415X18500015
  10. Kim DH, Jung HA, Sohn HS, Kim JW, Choi JS. Potential of icariin metabolites from Epimedium koreanum Nakai as antidiabetic therapeutic agents. Molecules. 2017 ; 22 : 986. https://doi.org/10.3390/molecules22060986
  11. Lee W, Nam JH, Cho HJ, Lee JY, Cho WK, Kim Y, We YM, Ma JY, Hoe HS. Epimedium koreanum Nakai inhibits PMA-induced cancer cell migration and invasion by modulating NF-${\kappa}B$/MMP-9 signaling in monomorphic malignant human glioma cells. Oncol Rep. 2017 ; 38 : 3619-31.
  12. Han YY, Song MY, Hwang MS, Hwang JH, Park YK, Jung HW. Epimedium koreanum Nakai and its main constituent icariin suppress lipid accumulation during adipocyte differentiation of 3T3-L1 preadipocytes. Chin J Nat Med. 2016 ; 14 : 671-6.
  13. Yuk SS. Effects of Epimedii Herba water extract on the inflammatory mediators in macrophage cells. Kyungwon University. 2009.
  14. Kim JK, Lee JE, Jung EH, Jung JY, Jung DH, Ku SK, Cho IJ, Kim SC. Hemistepsin A ameliorates acute inflammation in macrophages via inhibition of nuclear factor-${\kappa}B$ and activation of nuclear factor erythroid 2-related factor 2. Food Chem Toxicol. 2018 ; 111 : 176-88. https://doi.org/10.1016/j.fct.2017.11.014
  15. Hwangbo M, Jung JY, Ki SH, Park SM, Jegal KH, Cho IJ, Lee JH, Kang SH, Park SD, Ku SK, Kim SC, Zhao RJ, Jee SY, Kim YW. U-Bang-Haequi Tang: A Herbal Prescription that Prevents Acute Inflammation through Inhibition of NF-${\kappa}B$Mediated Inducible Nitric Oxide Synthase. Evid Based Complement Alternat Med. 2014 ; 2014 : 542825.
  16. Cho WK, Weeratunga P, Lee BH, Park JS, Kim CJ, Ma JY, Lee JS. Epimedium koreanum Nakai displays broad spectrum of antiviral activity in vitro and in vivo by inducing cellular antiviral state. Viruses. 2015 ; 7 : 352-77. https://doi.org/10.3390/v7010352
  17. Wang C, Feng L, Su J, Cui L, Dan Liu, Yan J, Ding C, Tan X, Jia X. Polysaccharides from Epimedium koreanum Nakai with immunomodulatory activity and inhibitory effect on tumor growth in LLCbearing mice. J Ethnopharmacol. 2017 ; 207 : 8-18. https://doi.org/10.1016/j.jep.2017.06.014
  18. Jin Q, Lee C, Lee JW, Yeon ET, Lee D, Han SB, Hong JT, Kim Y, Lee MK, Hwang BY. 2-Phenoxychromones and prenylflavonoids from Epimedium koreanum and their inhibitory effects on LPS-induced nitric oxide and interleukin-$1{\beta}$ production. J Nat Prod. 2014 ; 77 : 1724-8. https://doi.org/10.1021/np400831p
  19. Choi HJ, Eun JS, Park YR, Kim DK, Li R, Moon WS, Park JM, Kim HS, Cho NP, Cho SD, Soh Y. Ikarisoside A inhibits inducible nitric oxide synthase in lipopolysaccharide-stimulated RAW 264.7 cells via p38 kinase and nuclear factor-kappaB signaling pathways. Eur J Pharmacol. 2008 ; 601 : 171-8. https://doi.org/10.1016/j.ejphar.2008.09.032
  20. Guzik TJ, Korbut R, Adamek-Guzik T. Nitric oxide and superoxide in inflammation and immune regulation. J Physiol Pharmacol. 2003 ; 54 : 469-87.
  21. Schulze J, Weber K, Baranowsky A, Streichert T, Lange T, Spiro AS, Albers J, Seitz S, Zustin J, Amling M, Fehse B, Schinke T. p65-Dependent production of interleukin-$1{\beta}$ by osteolytic prostate cancer cells causes an induction of chemokine expression in osteoblasts. Cancer Lett. 2012 ; 317 : 106-13. https://doi.org/10.1016/j.canlet.2011.11.016
  22. Vila-del Sol V, Fresno M. Involvement of TNF and NF-${\kappa}B$ in the transcriptional control of cyclooxygenase-2 expression by IFN-$\gamma$ in macrophages. J Immunol. 2005 ; 174 : 2825-33. https://doi.org/10.4049/jimmunol.174.5.2825
  23. Qiao-Wen X, Richard W, and Carl N. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon-g and bacterial lipopolysaccharide. J Exp Med. 1993 ; 177 : 1779-84. https://doi.org/10.1084/jem.177.6.1779
  24. Rhodes KL, Golub SH, Economou JS. The regulation of the human tumor necrosis factor alpha promoter region in macrophage, T cell, and B cell lines. J Biol Chem. 1992 ; 267 : 22102-7.
  25. Libermann TA, Baltimore D. Activation of interleukin-6 gene expression through the NF-${\kappa}B$ transcription factor. Mol Cell Biol. 1990 ; 10 : 2327-34. https://doi.org/10.1128/MCB.10.5.2327
  26. Hoffmann A, Natoli G, Ghosh G. Transcriptional regulation via the Nf-Kappab signaling module. Oncogene. 2006 ; 25 : 6706-16. https://doi.org/10.1038/sj.onc.1209933
  27. Lai X, Ye Y, Sun C, Huang X, Tang X, Zeng X, Yin P, Zeng Y. Icaritin exhibits anti-inflammatory effects in the mouse peritoneal macrophages and peritonitis model. Int Immunopharmacol. 2013 ; 16 : 41-9. https://doi.org/10.1016/j.intimp.2013.03.025
  28. Xu CQ, Liu BJ, Wu JF, Xu YC, Duan XH, Cao YX, Dong JC. Icariin attenuates LPS-induced acute inflammatory responses involvement of PI3K Akt and NF-kappaB signaling pathway. Eur J Pharmacol. 2010 ; 642 : 146-53. https://doi.org/10.1016/j.ejphar.2010.05.012
  29. Endale M, Park SC, Kim S, Kim SH, Yang Y, Cho JY, Rhee MH. Quercetin disrupts tyrosinephosphorylated phosphatidylinositol 3-kinase and myeloid differentiation factor-88 association, and inhibits MAPK AP-1 and IKK NF-${\kappa}B$-induced inflammatory mediators production in RAW 264.7 cells. Immunobiology. 2013 ; 218 : 1452-67. https://doi.org/10.1016/j.imbio.2013.04.019