인간진피섬유아세포에서 6-Shogaol의 세포보호 및 항염증 효과 = Cytoprotective and Anti-inflmmatory Effects of 6-Shogaol on Human Dermal Fibroblasts

다국어 초록 (Multilingual Abstract)

Purpose: 6-Shogaol is one of the spicy flavor ingredients of ginger and hydrolysis product of gingerols. The purpose of this study is to investigate the potential of 6-shogaol as a natural cosmetic raw material by examining cell protection and inflammation inhibitory activity in human dermal fibroblasts (HDFs). Methods: To verify the cytoprotective effects of 6-shogaol, cell viability was measured by watersoluble tetrazolium salt (WST-1) assay. To verify the anti-inflammatory effects of 6-shogaol, we performed nuclear factor kappa-light-chain-enhancer of activated B cells (NFΚB) promoter luciferase assay to check the expression of NFΚB. Additionally, the expression of inflmmation related gene such as cyclooxygenase 2 (COX2), receptor for advanced glycation end product (RAGE), tumor necrosis factor alpha (TNFΑ), interleukin 6 (IL6), and interleukin 8 (IL8) mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Results: 6-Shogaol showed no toxicity in HDFs at concentrations of 5, 10, 20, and 40 μM, respectively and the cell viability was increased in a dose dependent manner. qRT-PCR analysis showed 6-shogaol treatment downregulated the expression of NFΚB, COX2, RAGE, TNFΑ , IL6, and IL8 in a dose dependent manner, resulting that 6-shogaol leads to protective activities against inflammatory responses in HDFs. Conclusion: As mentioned above, 6-shogaol restored cell viability and decreased the expression of inflammatory factors in a dose dependent manner. Consequently, these results suggest the possibility of 6-shogaol as cosmetic material preventing skin aging, through identifid functions on cytoprotection and anti-inflmmation.

국문 초록 (Abstract)

6-Shogaol은 생강(Zingiber officinale)의 매운맛을 내는 성분인 gingerols의 가수분해 산물로서, 본 연구에서는 6-shogaol 의 인간진피섬유아세포에서의 세포보호와 염증억제 효능을 검증함으로써 천연유래 화장품 원료로서의 사용가능성을 알아보고자 한다. 방법: 6-Shogaol의 세포보호 효과를 검증하기 위하여, water-soluble tetrazolium salt (WST-1) assay를 이용하여 세포생존율을 측정하였다. 6-Shogaol의 항염증 효과를 검증하기 위하여, nuclear factor kappa-light-chain-enhancer of activated B cells (NFΚB) promoter luciferase assay를 이용하여 염증반응 전사인자인 NFΚB의 발현을 측정하였으며 염증관련 유전자들인 cyclooxygenase 2 (COX2), receptor for advanced glycation end product (RAGE), tumor necrosis factor alpha (TNFΑ), interleukin 6 (IL6), interleukin 8 (IL8) 발현량을 quantitative real-time polymerase chain reaction (qRT-PCR)를 이용하여 측정하였다. 결과: 6-Shogaol은 5, 10, 20, 40 µM 각각의 농도에서 인간진피섬유아세포에 대한 독성을 보이지 않았으며 농도의존적으로 10 J/cm²의 ultraviolet A (UVA)에 의해 손상된 인간진피섬유아세포의 생존율을 회복시켰다. 대표적인 염증반응 전사인자인NFΚB의 promoter 활성이 6-shogaol에 의해 농도의존적으로 감소되었으며, 염증유발 유전자인 COX2, RAGEmRNA와 각종 염증관련 사이토카인인 TNFΑ, IL6, IL8 mRNA의 발현도 6-shogaol에 의해 농도의존적으로 감소되었다. 결론: 6-Shogaol은 세포보호 및 항염증에 대한 확인된 효능을 통해 피부노화를 예방하는 화장품 원료로서의 가능성을 보여줄 수 있으리라 사료된다

참고문헌 (Reference)

1 윤영민, "자외선(Ultravilet)이 피부 및 피부세포 내 신호전달체계에 미치는 영향" 한국피부과학연구원 11 (11): 417-426, 2013

2 차화준, "인간진피섬유아세포에서 코코아 주요성분인 Procyanidin B1이 JNK-AP1-TRE Axis를 통한 Matrix-Metalloprotease 1 발현조절에 미치는 영향" 한국피부과학연구원 13 (13): 761-765, 2015

3 Jaszewska E, "UVA-induced ROS generation inhibition by Oenothera paradoxa defatted seeds extract and subsequent cell death in human dermal fibroblasts" 126 : 42-46, 2013

4 Agarwal S, "Tumor necrosis factormediated cytotoxicity involves ADP-ribosylation" 140 : 4187-4192, 1988

5 Oh JH, "Thioredoxin secreted upon ultraviolet A irradiation modulates activities of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in human dermal fibroblasts" 423 : 218-226, 2004

6 Campisi J, "The role of cellular senescence in skin aging" 3 : 1-5, 1998

7 Nathan CF, "Secretory products of macrophages" 79 : 319-326, 1987

8 Baeuerle PA, "Pro-inflammatory signaling: last pieces in the NFkappaB puzzle?" 8 : 19-22, 1998

9 Greaves MW, "Pharmacologic agents released in ultraviolet inflammation studied by continuous skin perfusion" 54 : 365-367, 1970

10 Hruza LL, "Mechanisms of UV-induced inflammation" 100 : 35-41, 1993

11 Kim J, "Inhibition effect of Gynura procumbens extract on UV-B-induced matrixmetalloproteinase expression in human dermal fibroblas" 137 : 427-433, 2011

12 Longley BJ, "Immunology and the skin : current concepts" 548 : 225-232, 1988

13 Frydas S, "Generation of TNFα, IFNγ, IL-6, IL-4 and IL-10 in mouse serum from trichinellosis: effect of the anti-inflammatory compound 4-deoxypyridoxine (4-DPD)" 49 : 179-184, 1996

14 Ju Sin Kim, "Cytotoxic Components from the Dried Rhizomes of Zingiber officinale Roscoe" 대한약학회 31 (31): 415-418, 2008

15 Hirano T, "Complementary DNA for a novel human interleukin(BSF-2)that induces B lymphocytes to produce immunoglobulin" 324 : 73-76, 1986

16 Xie J, "Cellular signalling of the receptor for advanced glycation end products(RAGE)" 25 : 2185-2197, 2013

17 Yang WI, "Blocking the receptor for advanced glycation end product activation attenuates autoimmune myocarditis" 1197-1205, 2014

18 Gomez-Nicola D, "Blockade of IL-15 activity inhibits microglial activation through the NFκB, p38, and ERK1/2 pathways, reducing cytokine and chemokine release" 58 : 264-276, 2010

19 Mendes RA, "An overview on the expression of cyclooxygenase-2 in tumors of the head and neck" 45 : 124-128, 2009

20 Bak MJ, "6-Shogaol-rich extract from ginger up-regulates the antioxidant defense systems in cells and mice" 17 : 8037-8055, 2012

21 Ling H, "6-Shogaol, an active constituent of ginger, inhibits breast cancer cell invasion by reducing matrix metalloproteinase-9expression via blockade of nuclear factor-κB activation" 161 : 1763-1777, 2010

22 Huang HC, "-Shogaol inhibits α-MSH-induced melanogenesis through the acceleration of ERK and PI3K/Akt-mediated MITF degradation" 2014 : 842569-, 2014