JNK-AP1 기전을 통한 국화꽃 추출물의 MMP1 발현 변화 조절 = Extracts of Chrysanthemum indicum Linne Mediated Regulation of MMP1 via JNK-AP1 Pathway

국문 초록 (Abstract)

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.

다국어 초록 (Multilingual 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.

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