Ginsenoside Rg5:Rk1 attenuates TNF-α/IFN-γ-induced production of thymus- and activation-regulated chemokine (TARC/CCL17) and LPS-induced NO production via downregulation of NF-κB/p38 MAPK/STAT1 signaling in human keratinocytes and macrophages

  • Sungeun Ahn
  • Muhammad Hanif Siddiqi
  • Veronica Castro Aceituno
  • Shakina Yesmin Simu
  • Jinglou Zhang
  • Zuly Elizabeth Jimenez Perez
  • Yu-Jin Kim
  • Deok-Chun YangEmail author


Atopic dermatitis (AD) is a chronic skin disease that affects millions of people worldwide. Keratinocytes and macrophages are two cells types that play a pivotal role in the development of AD. These cells produced different chemokines and cytokines, especially thymus and activation-regulated chemokine (TARC/CCL17) and macrophage-derived chemokine (MDC/CCL22), as well as nitric oxide (NO) through inducible nitric oxide synthase (iNOS) and COX2 in response to stimulation by TNF-α/IFN-γ and lipopolysaccharide (LPS) respectively. These mediators are thought to be crucial regulators of the pathogenesis of AD. Although several natural compounds to treat AD have been studied, the effect of Rg5:Rk1 from Panax ginseng (P. ginseng) on AD has not yet been investigated. In this study, we evaluated the inhibitory effect of Rg5:Rk1 on TNF-α/IFN-γ stimulated keratinocytes (HaCaT cells) and LPS-stimulated macrophages (RAW 264.7 cells). Enzyme-linked immunosorbent assay (ELISA) data showed that pretreatment of HaCaT cells with Rg5:Rk1 significantly reduced the TNF-α/IFN-γ-induced increase in TARC/CCL17 expression in a dose-dependent manner. In addition, Rg5:Rk1 decreased LPS-mediated nitric oxide (NO) and reactive oxygen species (ROS) production in RAW 264.7 cells. A considerable reduction in messenger RNA (mRNA) expression of the aforementioned AD mediators was also observed. Pretreatment with Rg5:Rk1 attenuated the TNF-α/IFN-γ-induced phosphorylation of p38 MAPK, STAT1, and NF-κB/IKKβ in HaCaT cells. Together, these findings suggest that ginsenoside Rg5:Rk1 may have a potential anti-AD effect by suppressing NF-κB/p38 MAPK/STAT1 signaling.


Inflammation Atopic dermatitis Keratinocytes/macrophages TARC/CCL17 NF-κB p38 MAPK STAT1 



This research was supported by grants 312064-03-1-HD040 and 313038-03-1-SB010 from the Korea Institute of Planning and Evaluation for Technology (iPET) of the Ministry of Food, Agriculture, Forestry and Fisheries, Republic of Korea. The ginseng sample used in this study was provided by the Ginseng Bank of Kyung Hee University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


  1. Ahn S, Siddiqi MH, Noh H, Kim Y, Kim Y, Jin C, Yang D (2015) Anti-inflammatory activity of ginsenosides in LPS-stimulated RAW 264.7 cells. Sci Bull 60:773–784CrossRefGoogle Scholar
  2. Amarbayasgalan T, Takahashi H, Dekio I, Morita E (2013) Interleukin-8 content in the stratum corneum as an indicator of the severity of inflammation in the lesions of atopic dermatitis. Int Arch Allergy Immunol 160:63–74CrossRefPubMedGoogle Scholar
  3. Bao HY, Zhang J, Yeo SJ, Myung CS, Kim HM, Kim JM, Park JH, Cho J, Kang JS (2005) Memory enhancing and neuroprotective effects of selected ginsenosides. Arch Pharm Res 28:335–342CrossRefPubMedGoogle Scholar
  4. Barker JN, Palmer CN, Zhao Y, Liao H, Hull PR, Lee SP, Allen MH, Meggitt SJ, Reynolds NJ, Trembath RC, McLean WH (2007) Null mutations in the filaggrin gene (FLG) determine major susceptibility to early-onset atopic dermatitis that persists into adulthood. J Investig Dermatol 127:564–567CrossRefPubMedGoogle Scholar
  5. Coleman JW (2001) Nitric oxide in immunity and inflammation. Int Immunopharmacol 1:1397–1406CrossRefPubMedGoogle Scholar
  6. Fabricant DS, Farnsworth NR (2001) The value of plants used in traditional medicine for drug discovery. Environ Health Perspect 109(Suppl 1):69–75CrossRefPubMedPubMedCentralGoogle Scholar
  7. Gallicchio M, Rosa AC, Benetti E, Collino M, Dianzani C, Fantozzi R (2006) Substance P-induced cyclooxygenase-2 expression in human umbilical vein endothelial cells. Br J Pharmacol 147:681–689CrossRefPubMedPubMedCentralGoogle Scholar
  8. Gillis CN (1997) Panax ginseng pharmacology: a nitric oxide link? Biochem Pharmacol 54:1–8CrossRefPubMedGoogle Scholar
  9. Gros E, Bussmann C, Bieber T, Forster I, Novak N (2009) Expression of chemokines and chemokine receptors in lesional and nonlesional upper skin of patients with atopic dermatitis. J Allergy Clin Immunol 124:753–760 e1CrossRefPubMedGoogle Scholar
  10. Han EH, Hwang YP, Choi JH, Yang JH, Seo JK, Chung YC, Jeong HG (2011) Psidium guajava extract inhibits thymus and activation-regulated chemokine (TARC/CCL17) production in human keratinocytes by inducing heme oxygenase-1 and blocking NF-κB and STAT1 activation. Environ Toxicol Pharmacol 32:136–145CrossRefPubMedGoogle Scholar
  11. Harvima IT, Nilsson G (2011) Mast cells as regulators of skin inflammation and immunity. Acta Derm Venereol 91:640–649CrossRefGoogle Scholar
  12. Homey B, Steinhoff M, Ruzicka T, Leung DY (2006) Cytokines and chemokines orchestrate atopic skin inflammation. J Allergy Clin Immunol 118:178–189CrossRefPubMedGoogle Scholar
  13. Imai T, Nagira M, Takagi S, Kakizaki M, Nishimura M, Wang J, Gray PW, Matsushima K, Yoshie O (1999) Selective recruitment of CCR4-bearing Th2 cells toward antigen-presenting cells by the CC chemokines thymus and activation-regulated chemokine and macrophage-derived chemokine. Int Immunol 11:81–88CrossRefPubMedGoogle Scholar
  14. Jachak SM, Saklani A (2007) Challenges and opportunities in drug discovery from plants. Curr Sci 92:1251–1257Google Scholar
  15. Ju SM, Song HY, Lee SJ, Seo WY, Sin DH, Goh AR, Kang YH, Kang IJ, Won MH, Yi JS, Kwon DJ, Bae YS, Choi SY, Park J (2009) Suppression of thymus- and activation-regulated chemokine (TARC/CCL17) production by 1,2,3,4,6-penta-O-galloyl-beta-D-glucose via blockade of NF-kappaB and STAT1 activation in the HaCaT cells. Biochem Biophys Res Commun 387:115–120CrossRefPubMedGoogle Scholar
  16. Kakinuma T, Nakamura K, Wakugawa M, Mitsui H, Tada Y, Saeki H, Torii H, Asahina A, Onai N, Matsushima K (2001) Thymus and activation-regulated chemokine in atopic dermatitis: serum thymus and activation-regulated chemokine level is closely related with disease activity. J Allergy Clin Immunol 107:535–541CrossRefPubMedGoogle Scholar
  17. Kim DH, Chung JH, Yoon JS, Ha YM, Bae S, Lee EK, Jung KJ, Kim MS, Kim YJ, Kim MK (2013) Ginsenoside Rd inhibits the expressions of iNOS and COX-2 by suppressing NF-κB in LPS-stimulated RAW264. 7 cells and mouse liver. J Ginseng Res 37:54CrossRefPubMedPubMedCentralGoogle Scholar
  18. Kim SN, Ha YW, Shin H, Son SH, Wu SJ, Kim YS (2007) Simultaneous quantification of 14 ginsenosides in Panax ginseng CA Meyer (Korean red ginseng) by HPLC-ELSD and its application to quality control. J Pharm Biomed Anal 45:164–170CrossRefPubMedGoogle Scholar
  19. Kim YS, Young MR, Bobe G, Colburn NH, Milner JA (2009) Bioactive food components, inflammatory targets, and cancer prevention. Cancer Prev Res (Phila) 2:200–208CrossRefGoogle Scholar
  20. Kinne RW, Brauer R, Stuhlmuller B, Palombo-Kinne E, Burmester G (2000) Macrophages in rheumatoid arthritis. Arthritis Res 2:189–202CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kisseleva T, Bhattacharya S, Braunstein J, Schindler CW (2002) Signaling through the JAK/STAT pathway, recent advances and future challenges. Gene 285:1–24CrossRefPubMedGoogle Scholar
  22. Ko J, Lee JI, Munoz-Furlong A, Li XM, Sicherer SH (2006) Use of complementary and alternative medicine by food-allergic patients. Ann Allergy Asthma Immunol 97:365–369CrossRefPubMedGoogle Scholar
  23. Kwon DJ, Bae YS, Ju SM, Goh AR, Youn GS, Choi SY, Park J (2012) Casuarinin suppresses TARC/CCL17 and MDC/CCL22 production via blockade of NF-kappaB and STAT1 activation in HaCaT cells. Biochem Biophys Res Commun 417:1254–1259CrossRefPubMedGoogle Scholar
  24. Nakazato J, Kishida M, Kuroiwa R, Fujiwara J, Shimoda M, Shinomiya N (2008) Serum levels of Th2 chemokines, CCL17, CCL22, and CCL27, were the important markers of severity in infantile atopic dermatitis. Pediatr Allergy Immunol 19:605–613PubMedGoogle Scholar
  25. Pivarcsi A, Homey B (2005) Chemokine networks in atopic dermatitis: traffic signals of disease. Curr Allergy Asthma Rep 5:284–290CrossRefPubMedGoogle Scholar
  26. Pugliarello S, Cozzi A, Gisondi P, Girolomoni G (2011) Phenotypes of atopic dermatitis. J Dtsch Dermatol Ges 9:12–20PubMedGoogle Scholar
  27. Qi XF, Kim DH, Yoon YS, Li JH, Song SB, Jin D, Huang XZ, Teng YC, Lee KJ (2009) The adenylyl cyclase-cAMP system suppresses TARC/CCL17 and MDC/CCL22 production through p38 MAPK and NF-kappaB in HaCaT keratinocytes. Mol Immunol 46:1925–1934CrossRefPubMedGoogle Scholar
  28. Qi XF, Kim DH, Yoon YS, Song SB, Teng YC, Cai DQ, Lee KJ (2012) Bambusae caulis in liquamen suppresses the expression of thymus and activation-regulated chemokine and macrophage-derived chemokine in human keratinocytes due to antioxidant effect. Evid Based Complement Alternat Med 2012:617494PubMedPubMedCentralGoogle Scholar
  29. Sekine Y, Yumioka T, Yamamoto T, Muromoto R, Imoto S, Sugiyma K, Oritani K, Shimoda K, Minoguchi M, Akira S, Yoshimura A, Matsuda T (2006) Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J Immunol 176:380–389CrossRefPubMedGoogle Scholar
  30. Shimada Y, Takehara K, Sato S (2004) Both Th2 and Th1 chemokines (TARC/CCL17, MDC/CCL22, and Mig/CXCL9) are elevated in sera from patients with atopic dermatitis. J Dermatol Sci 34:201–208CrossRefPubMedGoogle Scholar
  31. Shin Y, Bae E, Kim D (2006) Inhibitory effect of ginsenoside Rg5 and its metabolite ginsenoside Rh3 in an oxazolone-induced mouse chronic dermatitis model. Arch Pharm Res 29:685–690CrossRefPubMedGoogle Scholar
  32. Siddiqi MH, Siddiqi MZ, Ahn S, Kang S, Kim YJ, Veerappan K, Yang DU, Yang DC (2014) Stimulative effect of ginsenosides Rg5:Rk1 on murine osteoblastic MC3T3-E1 cells. Phytother Res 28:1447–1455CrossRefPubMedGoogle Scholar
  33. Siddiqi MH, Siddiqi MZ, Kang S, Noh HY, Ahn S, Simu SY, Aziz MA, Sathishkumar N, Jimenez Perez ZE, Yang DC (2015) Inhibition of osteoclast differentiation by ginsenoside Rg3 in RAW264.7 cells via RANKL, JNK and p38 MAPK pathways through a modulation of Cathepsin K: an in silico and in vitro study. Phytother Res. doi: 10.1002/ptr.5374 PubMedGoogle Scholar
  34. Takeda K, Akira S (2001) Roles of Toll-like receptors in innate immune responses. Gene Cells 6:733–742CrossRefGoogle Scholar
  35. Tohyama M, Sayama K, Komatsuzawa H, Hanakawa Y, Shirakata Y, Dai X, Yang L, Tokumaru S, Nagai H, Hirakawa S, Sugai M, Hashimoto K (2007) CXCL16 is a novel mediator of the innate immunity of epidermal keratinocytes. Int Immunol 19:1095–1102CrossRefPubMedGoogle Scholar
  36. Zhuang Y, Lyga J (2014) Inflammaging in skin and other tissues—the roles of complement system and macrophage. Inflamm Allergy Drug Targets 13:153–161CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2015

Authors and Affiliations

  • Sungeun Ahn
    • 1
  • Muhammad Hanif Siddiqi
    • 2
  • Veronica Castro Aceituno
    • 1
  • Shakina Yesmin Simu
    • 2
  • Jinglou Zhang
    • 2
  • Zuly Elizabeth Jimenez Perez
    • 2
  • Yu-Jin Kim
    • 1
  • Deok-Chun Yang
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Oriental Medicinal Biotechnology, College of Life SciencesKyung Hee UniversityYonginRepublic of Korea
  2. 2.Ginseng Bank, Graduate School of Biotechnology, College of Life SciencesKyung Hee UniversityYonginRepublic of Korea
  3. 3.Department of Oriental Medicinal Materials and Processing, Graduate School of Biotechnology, College of Life ScienceKyung Hee UniversityYonginSouth Korea

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