브라질린의 RAW264.7 세포에서의 항염효과 = Anti-inflammatory Effect of Brazilin in RAW264.7 Cells

다국어 초록 (Multilingual Abstract)

Purpose: This study aimed to determine the skin anti-inflammatory effect of brazilin on RAW264.7 cells. Methods: To determine anti-inflammatory effects of brazilin on RAW264.7 cells, we measured cell viability, gene expressions, and prostaglandin E 2 (PGE 2 )/nitric oxide (NO) production. Results: Cells were treated with brazilin to confirm cytotoxicity; no toxicity was observed when cells were treated at a concentration of ≤1 μg/mL. In RAW264.7 cells, the number of intracellular granule were increased after lipopolysaccharide (LPS) treatment; however, brazilin treatment decreased intracellular granules similar to that in the control group. In this research, we elucidated the anti-inflammatory effects of brazilin on the mRNA expression of interleukin 1 alpha (IL1α), interleukin 1 beta (IL1β), interleukin 6 (IL6), and tumor necrosis factor alpha (TNFα) and PGE 2 /NO production. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) showed that brazilin administration increased the mRNA levels of IL1α /IL1β /IL6 /TNFα genes, and analysis of PGE 2 /NO also revealed that brazilin induces anti-inflammatory effects. Conclusion: We identified the anti-inflammatory effect of brazilin, and found that brazilin can be a potent cosmetic ingredient for anti-inflammatory agent for atopic dermatitis. Based on this result, we believe that brazilin has the potential to be used in not only cosmetics, but also medicine or healthcare food.

국문 초록 (Abstract)

목적: 본 연구는 브라질린에 의한 RAW264.7 세포의 항염 효과를 확인하기 위하여 수행되었다. 방법: 브라질린이 RAW264.7세포에 항염효과를 나타내는지 확인하기 위하여, CCK-8 실험법을 통한 세포생존률 측정, 실시간 유전자 중합효소 연쇄반응을 통한 싸이토카인의 유전자 발현양상, 효소결합면역흡착검사법을 통한 PGE 2 와 산화질소의 생성 정도를 확인하였다. 결과: RAW264.7 세포에 브라질린을 처리하여 세포독성을 확인한 결과, 1 μg/mL 이하의 농도로 처리시 독성을 나타내지 않음을 확인할 수 있었다.
RAW264.7 세포는 LPS 자극에 의해 세포질 내 소과립이 증가되나 브라질린을 처리하면 대조군과 유사하게 세포질 내 소과립이 감소하는 경향을 보였다. 본 연구를 통해 브라질린이 인터류킨1α (interleukin 1α, IL1α), 인터류킨1β (interleukin 1β, IL1β), 인터류킨 6 (interleukin 6, IL6), tumor necrosis factor α (TNFα) 유전자의 발현을 감소시킴을 확인할 수 있었다. 또한 산화질소의 생성을 감소시키고 PGE 2 의 생성을 감소시킴을 통해 브라질린의 항염효과를 확인할 수 있었다. 결론: 본 연구를 통해 브라질린의 항염효과를 확인하였고, 브라질린이 항염 및 아토피를 타겟으로 하는 화장품 원료로서의 가능성을 제시하였다. 브라질린이 피부미용, 화장품, 건강식품 등의 산업 등에도 응용될 수 있을 것으로 여겨지며 그 가능을 확인하기 위해 브라질린이 염증세포에 미치는 기전연구가 필요할 것으로 보인다.

참고문헌 (Reference)

1 임도연, "화피의 휘발성 향기성분 조성과 주요성분인 Benzaldehyde의 생리활성" 한국피부과학연구원 12 (12): 155-162, 2014

2 손명수, "홍삼에서 추출한 오일의 항산화 및 보습활성" 한국피부과학연구원 11 (11): 489-494, 2013

3 정금선, "한국산 갈퀴덩굴속(Galium L.)의 세포분류학적 연구" 한국식물분류학회 39 (39): 42-47, 2009

4 백영미, "구판의 LPS로 유도된 염증 반응 억제 효과" 한의병리학회 26 (26): 26-34, 2012

5 이해진, "加味消風散이 염증 및 아토피피부염 동물병태에 미치는 영향" 한의병리학회 28 (28): 146-153, 2014

6 Kirkwood TB, "Understanding the odd science of aging" 120 : 437-447, 2005

7 Akira S, "Toll-like receptor signaling" 4 : 499-511, 2004

8 Noman AS, "Thalidomide inhibits lipopolysaccharide-induced tumor necrosis factor-alpha production via down-regulation of MyD88 expression" 15 : 33-41, 2009

9 Gomez PF, "Resolution of inflammation: prostaglandin E2dissociates nuclear trafficking of individual NF-kappa B subunits (p65, p50) in stimulated rheumatoid synovial fibroblasts" 175 : 6924-6930, 2005

10 Talwar HS, "Reduced type I and type III procollagens in photodamaged adult human skin" 105 : 285-290, 1995

11 Xu X, "Punicalagin induces Nrf2/HO-1 expression via upregulation of PI3K/AKT pathway and inhibits LPSinduced oxidative stress in RAW264.7 macrophages" 2015 : 380218-, 2015

12 Harris SG, "Prostaglandins as modulators of immunity" 23 : 144-150, 2002

13 Albina JE, "Nitric oxide in inflammation and immunity" 3 : 46-64, 1995

14 Zhang Y, "Neutrophil recruitment by tumor necrosis factor from mast cells in immune complex peritonitis" 258 : 1957-1959, 1992

15 반주연, "Microarray Analysis of Gene Expression Profiles in Response to Treatment with Melatonin in Lipopolysaccharide Activated RAW 264.7 Cells" 대한약리학회 15 (15): 23-29, 2011

16 Fujiwara N, "Macrophages in inflammation" 4 : 281-286, 2005

17 Hirohashi N, "Low-dose lipopolysaccharide (LPS)pretreatment of mouse macrophages modulates LPSdependent interleukin-6 production in vitro" 64 : 1011-1015, 1996

18 Gabay C, "Interleukin-6 and chronic inflammation" 8 : S3-, 2006

19 Yun KJ, "Inhibition of LPSinduced NO and PGE2 production by asiatic acid via NF-kB inactivation in RAW 264.7 macrophages:possible involvement of the IKK and MAPK pathways" 8 : 431-441, 2008

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

21 Roitt I, "Immunology sixth edition" Mosby 119-441, 2001

22 Lord PC, "Expression of interleukin-1 alpha and beta genes by human blood polymorphonuclear leukocytes" 87 : 1312-1321, 1991

23 Hong CH, "Evaluation of natural products on inhibition of inducible cyclooxygenase (COX-2) and nitric oxide synthase (iNOS) in cultured mouse macrophage cells" 83 : 153-159, 2002

24 Jones DL, "Emerging models and paradigms for stem cell ageing" 13 : 506-512, 2011

25 Hwang GS, "Effects of brazilin on the phospholipase A2 activity and changes of intracellular free calcium concentration in rat platelets" 21 : 774-778, 1998

26 Kim S, "Compound K induces expression of hyaluronan synthase 2 gene in transformed human keratinocytes and increases hyaluronan in hairless mouse skin" 316 : 348-355, 2004

27 Namazi MR, "Cetirizine and allopurinol as novel weapons against cellular autoimmune disorders" 4 : 349-353, 2004

28 Horwood NJ, "Bruton's tyrosine kinase is required for TLR2 and TLR4-induced TNF, but not IL-6, production" 176 : 3635-3641, 2006

29 Moon CK, "Brazilin protects cultured rat hepatocytes from BrCCl3-induced toxicity" 15 : 81-91, 1992

30 Hwang HS, "Brazilin and Caesalpinia sappan L. extract protect epidermal keratinocytes from oxidative stress by inducing the expression of GPX7" 16 : 203-209, 2018

31 Puchtler H, "Application of current chemical concepts to metal-hematein and -brazilein stains" 85 : 353-364, 1986

32 Lawrence T, "Anti-inflammatory lipid mediators and insights into the resolution of inflammation" 2 : 787-795, 2002

33 Van Triel JJ, "Allergic inflammation in the upper respiratory tract of the rat upon repeated inhalation exposure to the contact allergen dinitrochlorobenzene (DNCB)" 269 : 73-80, 2010

34 Park JE, "Alkaloids from Chelidonium majus and their inhibitory effects on LPSinduced NO production in RAW264.7 cells" 21 : 6960-6963, 2011