JNK-AP1 기전을 통한 국화꽃 추출물의 MMP1 발현 변화 조절
Extracts of Chrysanthemum indicum Linne Mediated Regulation of MMP1 via JNK-AP1 Pathway
1
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
목적: 국화꽃은 다양한 항산화 물질을 내포하고 있어 한국, 중국 일본 등지에서 예로부터 감기, 두통, 어지러움, 고혈압 등에 사용되어 왔다. 그러나 지금까지 피부 진피에 국화꽃이 미치는 영향에 대해서는 잘 알려지지 않았다. 따라서 본 연구진은 실험을 통해, 국화꽃이 진피층의 세포외기질을 조절하여 피부노화를 경감시킬 수 있음을 제시하였다. 방법: 실험에 사용한 국화꽃 추출물은 건조한 국화꽃을 분쇄하여 70% ethanol을 사용하여 침출하였고, MTT assay를 통해 세포독성실험과 이후 실험에 사용할 농도를 정하였다. qRT-PCR을 이용하여 MMP1의 mRNA 발현 변화를 확인하였으며, pGL-TRE luciferase reporter vector를 활용하여 AP1의 promoter 활성을 측정하였다. JNK와 p-JNK의 단백질 발현은 western blot을 통해 확인하였다. 결과: 실험을 통해 국화꽃 추출물이 산화적 스트레스가 유도하는 MMP1 발현을 감소시키는 것을 증명하였으며 더불어, 이러한 현상이 국화꽃 추출물이 MMP1의 상위조절인자인 AP1의 전사활성을 조절하는 것으로부터 야기됨을 밝혔다. 마지막으로는 AP1이 JNK의 인산화를 저해하는 기작으로부터 조절되는 것을 증명하여 종합적으로 국화꽃 추출물이 JNK-AP1 경로를 통해 MMP1의 발현을 조절하는 것을 입증하였다. 결론: 본 연구를 통해 국화꽃 추출물이 노화와 주름생성을 경감시킬 수 있는 잠재적인 화장품 원료로서의 가능성을 확인하였다.
Abstract
Purpose: Chrysanthemum morifolium flower, Chrysanthemum indicum Linne (C. indicum Linne), contains various antioxidants which are used to care cold, headache, dizziness, and hypertension in Korea, China, and Japan. However, in dermis, effects of C. indicum Linne are not understood, currently. Thus, in present study, we suggest an evidence that C. indicum Linne decreases skin aging by regulating extracellular matrix of dermis. Methods: Dried C. indicum Linne was extracted with 70% ethanol and concentrations of this experiment were determined by cytotoxicity with MTT assay. Matrix metalloprotease 1 (MMP1) mRNA expression was confirmed by qRT-PCR and activity of AP1 promoter was measured with pGL-TRE luciferase reporter vector. Moreover, protein expression of JNK and p-JNK was assessed by western blot. Results: As shown results, we demonstrated that C. indicum Linne extracts decreased MMP1 expression induced by oxidative stress. In addition, C. indicum Linne extracts repressed MMP1 transcription by regulating AP1 transcriptional activity. Lastly, we demonstrated AP1 was regulated by blocking phosphorylation of JNK. Overall, C. indicum Linne extracts regulates MMP1 expression using JNK-AP1 pathway. Conclusion: C. indicum Linne extracts has potential to reduce aging and formation of wrinkle and to use as a cosmetic ingredient.
참고문헌 (24)
- An DG, "Illustrated book of Korean medicinal herbs", pp.1~115 (1998), Kyohak Publishing
- Angel P, "Function and regulation of AP-1 subunits in skin physiology and pathology", Oncogene, 20, pp.2413~2423 (2001)
- Bateman JF, "Extracellular matrix", pp.22~67 (1996), Harwood Academic Publishers
- Chung HY, "Dietary modulation of prostanoid synthesis in the aging process: role of cyclooxygenase-2", Mechanisms of Ageing and Development, 111, pp.97~106 (1999)
- Chung JH, "Regulations of collagen synthesis by ascorbic acid, transforming growth factor-beta and interferon-gamma in human dermal fibroblasts cultured in three-dimensional collagen gel are photoaging- and aging-independent", Journal of Dermatological Science, 15, pp.188~200 (1997)
- Coulomb B, "The collagen lattice: a model for studying the physiology, biosynthetic function and pharmacology of the skin", British Journal of Dermatology, 111, pp.83~87 (1984)
- Dasgupta J, "Reactive oxygen species control senescence-associated matrix metalloproteinase-1 through c-Jun-N-terminal kinase", Journal of Cellular Physiology, 225, pp.52~62 (2010)
- Dy LC, "Augmentation of ultraviolet B radiation-induced tumor necrosis factor production by the epidermal platelet-activating factor receptor", The Journal of Biological Chemistry, 274, pp.26917~26921 (1999)
- Fisher GJ, "Mechanisms of photoaging and chronological skin aging", Archives of Dermatology, 138, pp.1462~1470 (2002)
- Fisher GJ, "Molecular mechanisms of photoaging and its prevention by retinoic acid: ultraviolet irradiation induces MAP kinase signal transduction cascades that induce Ap-1-regulated matrix metalloproteinases that degrade human skin in vivo", Journal of Investigative Dermatology Symposium Proceedings, 3, pp.61~68 (1998)
- Fisher GJ, "Pathophysiology of premature skin aging induced by ultraviolet light", The New England Journal of Medicine, 337, pp.1419~1428 (1997)
- Gross S, "Inactivation of protein-tyrosine phosphatases as mechanism of UV-induced signal transduction", The Journal of Biological Chemistry, 274, pp.26378~26386 (1999)
- Karin M, "The regulation of AP-1 activity by mitogen-activated protein kinases", The Journal of Biological Chemistry, 270, pp.16483~16486 (1995)
- 이현정, "국화차를 포함하는 허브차의 CCl_4로 유도된 간세포손상 보호 및 항유전독성 효과", 한국식품영양과학회지, 40(1), pp.78~83 (2011), 한국식품영양과학회
- Leppä S, "Differential regulation of c-Jun by ERK and JNK during PC12 cell differentiation", The EMBO Journal, 17, pp.4404~4413 (1998)
- Nam SH, "Antibacterial activities of extracts from Chrysanthemum boreale M", Applied Biological Chemistry, 38, pp.269~272 (1995)
- Park KK, "Finding of TRE (TPA responsive element) in the sequence of human taurine transporter promoter", Advances in Experimental Medicine and Biology, 526, pp.159~166 (2003)
- Park NY, "Optimization of extraction conditions for ethanol extracts from Chrysanthemum morifolium by response surface methodology", Korean Journal of Food Science and Technology, 30, pp.1189~1196 (1998)
- Poulalhon N, "Modulation of collagen and MMP-1 gene expression in fibroblasts by the immunosuppressive drug rapamycin. a direct role as an antifibrotic agent?", The Journal of Biological Chemistry, 281, pp.33045~33052 (2006)
- Risse G, "Asymmetrical recognition of the palindromic AP1 binding site (TRE) by fos protein complexes", The EMBO Journal, 8, pp.3825~3832 (1989)
- Rittié L, "UV-light-induced signal cascades and skin aging", Ageing Research Reviews, 1, pp.705~720 (2002)
- Uitto J, "Connective tissue biochemistry of the aging dermis. age-related alterations in collagen and elastin", Dermatologic Clinics, 4, pp.433~446 (1986)
- Varani J, "Inhibition of type I procollagen synthesis by damaged collagen in photoaged skin and by collagenase-degraded collagen in vitro", The American Journal of Pathology, 158, pp.931~942 (2001)
- Westermarck J, "Enhancement of fibroblast collagenase-1 (MMP-1) gene expression by tumor promoter okadaic acid is mediated by stress-activated protein kinases Jun N-terminal kinase and p38", Matrix Biology, 17, pp.547~557 (1998)