[논문]송라 추출물의 세포 수준에서 항노화 및 모유두세포 활성화 효과

김민정; 최원영; 심현우; 신은진; 이정노; 박성민; 유화선

doi:10.15230/scsk.2024.50.1.37

송라 추출물의 세포 수준에서 항노화 및 모유두세포 활성화 효과
In Vitro Anti-aging and Hair Follicle Dermal Papilla Cells Activation Effects of Usnea diffracta Vain Extract 원문보기

大韓化粧品學會誌 = Journal of the society of cosmetic scientists of Korea, v.50 no.1, 2024년, pp.37 - 48

김민정 ((주)코씨드바이오팜 피부사랑임상연구센터) , 최원영 ((주)코씨드바이오팜 피부사랑임상연구센터) , 심현우 ((주)코씨드바이오팜 피부사랑임상연구센터) , 신은진 ((주)코씨드바이오팜 피부사랑임상연구센터) , 이정노 ((주)코씨드바이오팜 피부사랑임상연구센터) , 박성민 ((주)코씨드바이오팜 피부사랑임상연구센터) , 유화선 ((주)코씨드바이오팜 피부사랑임상연구센터)

초록
AI-Helper

송라(Usnea diffracta Vain.)는 송라속(Usnea)에 속하는 지의류 중 하나이며, 이전 연구에서 항산화, 항균, 항염, 항종양 및 심혈관 보호 등의 약리학적 활성이 보고되어 있으나 피부 및 모발에서의 효능은 잘 알려져 있지 않다. 따라서 본 연구에서는 세포 수준에서 송라 추출물(UDE)의 항노화 및 모유두세포 증식에 대한 효과를 검증하였다. 실험 결과, 송라 추출물은 인간 섬유아세포에서 UVA에 의해 증가된 MMP-1의 발현 및 상위기전인 MAPKs (ERK, p38, JNK)와 AP-1 (c-Fos, c-Jun)의 활성을 유의적으로 감소하는 것을 확인하였다. 또한, 송라 추출물은 인간 모유두세포의 증식을 유의하게 증가시켰으며, 모발 성장인자인 VEGF 및 KGF의 mRNA 발현을 유의하게 증가시켰다. 이로 인하여, 모발 증식 및 성장인자의 발현에 관여하는 ERK/CREB의 인산화를 농도 의존적으로 증가시켰다. 송라 추출물의 주성분 확인을 위해 송라 추출물을 농축 후 Prep-LC를 이용하여 main peak로 나타난 분획을 분리 정제하였고, NMR 및 Mess 분석을 통해 diffractaic acid로 동정하였다. Diffractaic acid는 인간 섬유아세포에서 UVA에 의해 증가된 MMP-1의 발현을 유의적으로 감소시켰으며, 인간 모유두세포의 증식을 농도 의존적으로 증가시켰다. 이를 통해 송라 추출물은 항노화 및 모유두세포 활성 증가 효능을 갖는 화장품 천연소재로서의 활용 가능성을 입증하였다.

Abstract ▼ AI-Helper

Songla (Usnea diffracta Vain.) is one of the lichens belonging to the genus Usnea, and pharmacological activities such as antioxidant, antimicrobial, anti-inflammatory, anti-tumor and cardiovascular protection have been reported in previous studies, but its efficacy in skin and hair is not well known. In this study, the effect of Usnea diffracta extract (UDE) on anti-aging and dermal papilla cell proliferation was verified in vitro. As a result of the experiment, it was confirmed that the UDE significantly reduced the expression of MMP-1 and the activity of MAPKs (ERK, p38, JNK) and AP-1 (c-Fos, c-Jun), which were increased by UVA in HDFn. In addition, the UDE significantly increased the proliferation of HFDPC and significantly increased the mRNA expression of VEGF and KGF, which are hair growth factors. Accordingly, the phosphorylation of ERK/CREB involved in hair proliferation and expression of growth factors was increased in a concentration-dependent manner. The main component represented by the main peak was separated and purified using Prep LC by concentrating the UDE, which was confirmed as diffractaic acid through NMR and Mess analysis. Isolated diffractaic acid significantly reduced the expression of MMP-1 increased by UVA in HDFn and increased the proliferation of HFDPC in a concentration-dependent manner. The result suggest that UDE proved its usability as a natural cosmetic material with anti-aging and dermal papilla cell activation effects.

주제어

표/그림 (9)

$Figure 1. The flowchart of Usnea diffracta extracts liquid-liquid separation.$ 그림 Figure 1. The flowchart of Usnea diffracta extracts liquid-liquid separation.
표 Table 1. Polymerase Chain Reaction (PCR) Primer Sequences
$Figure 2. Effect of Usnea diffracta extracts (UDE) on MMP-1 expression in HDFn cells. (A) Cells were treated with UDE for 24 h. Cell viability was determined using MTT assay. (B) Cells were irradiated with UVA (2 J/cm2) followed by treatment various concentrations with UDE for 6 h. mRNA and protein expressions of MMP-1 were examined using RT-PCR and western blot analysis. Band intensities relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. ###p < 0.001 represent significant difference compared to control and **p < 0.01 and ***p < 0.001 compared to UVA treated group.$ 그림 Figure 2. Effect of Usnea diffracta extracts (UDE) on MMP-1 expression in HDFn cells. (A) Cells were treated with UDE for 24 h. Cell viability was determined using MTT assay. (B) Cells were irradiated with UVA (2 J/cm²) followed by treatment various concentrations with UDE for 6 h. mRNA and protein expressions of MMP-1 were examined using RT-PCR and western blot analysis. Band intensities relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. ^###p < 0.001 represent significant difference compared to control and **p < 0.01 and ***p < 0.001 compared to UVA treated group.
$Figure 3. Effect of Usnea diffracta extracts (UDE) on MAPK/AP-1 signaling pathway expression in HDFn cells. Cells were irradiated with UVA (2 J/cm2) followed by treatment various concentrations with UDE for 20 min. Protein expressions of (A) MAPK and (B) AP-1 were examined using western blot analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. ###p < 0.001 represent significant difference compared to control and *p < 0.05 and ***p < 0.001 compared to UVA treated group.$ 그림 Figure 3. Effect of Usnea diffracta extracts (UDE) on MAPK/AP-1 signaling pathway expression in HDFn cells. Cells were irradiated with UVA (2 J/cm²) followed by treatment various concentrations with UDE for 20 min. Protein expressions of (A) MAPK and (B) AP-1 were examined using western blot analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. ^###p < 0.001 represent significant difference compared to control and *p < 0.05 and ***p < 0.001 compared to UVA treated group.
$Figure 4. Effect of Usnea diffracta extracts (UDE) on Cell proliferation and growth factor expression in HFDPC cells. (A) HFDPC cells were treated with UDE for 48 h. Cell proliferation was determined using MTT assay. (B) HFDPC cells were treated with UDE for 6 h. mRNA expressions of VEGF and KGF was examined using RT-PCR analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. *p < 0.05 and ***p < 0.001 represent significant difference compared to control.$ 그림 Figure 4. Effect of Usnea diffracta extracts (UDE) on Cell proliferation and growth factor expression in HFDPC cells. (A) HFDPC cells were treated with UDE for 48 h. Cell proliferation was determined using MTT assay. (B) HFDPC cells were treated with UDE for 6 h. mRNA expressions of VEGF and KGF was examined using RT-PCR analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. *p < 0.05 and ***p < 0.001 represent significant difference compared to control.
$Figure 5. Effect of Usnea diffracta extracts (UDE) on ERK/CREB signaling pathway expression in HFDPC cells. Cells were treated with UDE for 30 min. Protein expressions of ERK, phospho-ERK, CREB, and phospho-CREB were examined using western blot analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 represent significant difference compared to control.$ 그림 Figure 5. Effect of Usnea diffracta extracts (UDE) on ERK/CREB signaling pathway expression in HFDPC cells. Cells were treated with UDE for 30 min. Protein expressions of ERK, phospho-ERK, CREB, and phospho-CREB were examined using western blot analysis and relative expression levels were calculated using the ImageJ 1.47 software. All data are the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 represent significant difference compared to control.
$Figure 6. HPLC chromatogram data of diffractaic acid. (A) shows HPLC chromatogram of diffractaic acid and usnic acid and (B) shows HPLC chromatogram of Usnea diffracta extracts (UDE).$ 그림 Figure 6. HPLC chromatogram data of diffractaic acid. (A) shows HPLC chromatogram of diffractaic acid and usnic acid and (B) shows HPLC chromatogram of Usnea diffracta extracts (UDE).
$Figure 8. Effect of diffractaic acid on MMP-1 expression and hair papilla cells proliferation. (A) HDFn cells were irradiated with UVA (2 J/cm2) followed by treatment various concentrations with diffractaic acid for 6 h. mRNA and protein expressions of MMP-1 were examined using RT-PCR and western blot analysis. Band intensities relative expression levels were calculated using the ImageJ 1.47 software. (B) HFDPC cells were treated with diffractaic acid for 48 h. Cell proliferation was determined using MTT assay. All data are the mean ± SD of three independent experiments. ###p < 0.001 represent significant difference compared to control and **p < 0.01 and ***p < 0.001 compared to UVA treated group.$ 그림 Figure 8. Effect of diffractaic acid on MMP-1 expression and hair papilla cells proliferation. (A) HDFn cells were irradiated with UVA (2 J/cm²) followed by treatment various concentrations with diffractaic acid for 6 h. mRNA and protein expressions of MMP-1 were examined using RT-PCR and western blot analysis. Band intensities relative expression levels were calculated using the ImageJ 1.47 software. (B) HFDPC cells were treated with diffractaic acid for 48 h. Cell proliferation was determined using MTT assay. All data are the mean ± SD of three independent experiments. ^###p < 0.001 represent significant difference compared to control and **p < 0.01 and ***p < 0.001 compared to UVA treated group.
$Figure 7. The Chemical structure of diffractaic acid isolated from Usnea diffracta Vain.$ 그림 Figure 7. The Chemical structure of diffractaic acid isolated from Usnea diffracta Vain.

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저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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