[논문]CCD-986sk 세포 내 주박 분획물의 항주름 효능

장영아; 이혜정

doi:10.12925/jkocs.2024.41.2.508

CCD-986sk 세포 내 주박 분획물의 항주름 효능
Anti-wrinkle effects of solvent fractions from Jubak on CCD-986sk

Journal of the Korean Applied Science and Technology = 한국응용과학기술학회지, v.41 no.2, 2024년, pp.508 - 519

장영아 (대구한의대학교 화장품공학부) , 이혜정 (덕성여자대학교 약학대학)

초록
AI-Helper

본 연구에서는 주박의 기능성 화장품 소재로서의 활용 가능성을 평가하기 위해 분획물에 따른 항산화 활성평가와 인간섬유아세포인 CCD-986sk 세포에서의 주름효능 평가를 진행하였다. ABTS⁺ radical 소거능 측정을 통해 항산화 활성을 확인한 결과, 주박 Ethyl Acetate 분획물 1,000 ㎍/ml 농도에서 75.5%로 나타나 분획 용매 중 가장 높은 항산화 활성을 보였다. Elastase, Collagenase 저해 활성 측정을 통해 주름 개선 효과를 확인하였으며, 두 실험 결과 모두 주박 Ethyl Acetate 분획물이 1,000 ㎍/ml 농도에서 가장 높은 효능을 나타내었다. UVB로 유도된 CCD-986sk 세포 내 pro-collagen type I의 합성율을 측정한 결과 주박은 동일농도에서 20 ㎍/ml에서 Ethyl Acetate, Water, Acetonitrile, Hexan 분획물 순으로 높은 효능을 보였다. 콜라겐 분해효소인 MMP-1의 저해률을 측정한 결과 4가지 용매 분획몰은 20 ㎍/ml에서 모두 70% 이상의 효능을 나타내었다. Real time PCR 실험에서 pro-collagen type I, MMP-1 및 MMP-3의 mRNA 발현량을 측정한 결과 UVB 단독군 대비 Jubak 추출물을 함께 처리했을 때 pro-collagen type I의 단백질 발현량이 증가되었고, MMP-1과 MMP-3의 mRNA 발현량은 감소하는 것을 확인하였으며 대조군(EGCG) 다음으로 Ethyl Acetate 추출물이 주름 개선에 가장 효능이 우수하였다. 이상의 결과로 주박의 용매 분획물 중 Ethyl Acetate 분획물은 UVB 자극에 의한 광노화에 대한 항주름 효과가 있음을 확인하여 화장품의 천연물 소재로서의 이용이 기대된다.

Abstract ▼ AI-Helper

In this study, in order to evaluate the possibility of using Jubak as a functional cosmetic material, evaluation of antioxidant activity according to fractions and anti-wrinkle efficacy in CCD-986sk cells, a human fibroblast, were conducted. As a result of confirming the antioxidant activity by measuring ABTS+ radical scavenging ability, Jubak's Ethyl Acetate fractions was found to be 75.5% at a concentration of 1,000 ㎍/ml, showing the highest antioxidant activity among the extraction solvents. The wrinkle improvement effect was confirmed by measuring the inhibitory activity of elastase and collagenase, and in both test results, Jubak's Ethyl Acetate fractions showed the highest efficacy at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I in CCD-986sk cells induced by UVB, Jubak showed the highest efficacy in the order of Ethyl Acetate, Water, Acetonitrile, and Hexan fractions at the same concentration of 20 ㎍/ml. As a result of measuring the inhibition rate of MMP-1, a collagen degrading enzyme, all four solvent fractions showed an efficacy of more than 70% at 20 ㎍/ml. As a result of measuring the mRNA expression levels of pro-collagen type I, MMP-1, and MMP-3 in a real-time PCR experiment, the protein expression level of pro-collagen type I increased when treated with Jubak fractions compared to the UVB group alone. The mRNA expression levels of MMP-1 and MMP-3 were confirmed to be decreased, and Ethyl Acetate fractions was the most effective in improving wrinkles after the control group (EGCG). As a result, it was confirmed that the Ethyl Acetate fractions among Jubak's solvent fractions has an anti-wrinkle effect against photoaging caused by UVB stimulation, and is expected to be used as a natural material for cosmetics.

주제어

표/그림 (8)

$Fig. 1. ABTS<sup>+</sup> radical scavenging activity of Jubak fractions. The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-k) above the bars are significantly different at p<0.05.$ 그림 Fig. 1. ABTS⁺ radical scavenging activity of Jubak fractions. The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-k) above the bars are significantly different at p<0.05.
표 Table 1. Primer sequences.
$Fig. 2. Elastase inhibition activity of Jubak fractions. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-m) above the bars are significantly different at p<0.05.$ 그림 Fig. 2. Elastase inhibition activity of Jubak fractions. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-m) above the bars are significantly different at p<0.05.
$Fig. 3. Collagenase inhibition activity of Jubak fractions. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-l) above the bars are significantly different at p<0.05.$ 그림 Fig. 3. Collagenase inhibition activity of Jubak fractions. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. Statistical analysis was performed using Duncan's multiple range test. Means with different letters (a-l) above the bars are significantly different at p<0.05.
$Fig. 4. Cell viability of Jubak fractions in CCD-986sk fibroblast cells. The cells were treated with concentrations (0, 5, 10, 20, 40. 80, 160 μg/ml) of Jubak extract. Cell viability was measured using the MTT assay. The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the extract treated only group. *p< 0.05).$ 그림 Fig. 4. Cell viability of Jubak fractions in CCD-986sk fibroblast cells. The cells were treated with concentrations (0, 5, 10, 20, 40. 80, 160 μg/ml) of Jubak extract. Cell viability was measured using the MTT assay. The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the extract treated only group. *p< 0.05).
$Fig. 5. Effect of Jubak fractions on UVB-induced pro-collagen type I production in CCD-986sk fibroblast cells. The cells were treated with Jubak fractions (10, 20 μg/ml) and EGCG (10 μg/ml). The production of pro-collagen was determined by ELISA kit. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the extract treated only group.. *p< 0.05).$ 그림 Fig. 5. Effect of Jubak fractions on UVB-induced pro-collagen type I production in CCD-986sk fibroblast cells. The cells were treated with Jubak fractions (10, 20 μg/ml) and EGCG (10 μg/ml). The production of pro-collagen was determined by ELISA kit. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the extract treated only group.. *p< 0.05).
$Fig. 6. Effect of Jubak fractions on UVB-induced MMP-1 expression in CCD-986sk fibroblast cells. The cells were treated with Jubak fractions (10, 20μg/ml) and EGCG (10 μg/ml). The expression of MMP-1 was determined by ELISA kit. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the stimulus only group. *p< 0.05).$ 그림 Fig. 6. Effect of Jubak fractions on UVB-induced MMP-1 expression in CCD-986sk fibroblast cells. The cells were treated with Jubak fractions (10, 20μg/ml) and EGCG (10 μg/ml). The expression of MMP-1 was determined by ELISA kit. EGCG: Epigallocatechin gallate (Positive control). The result was presented as mean ± S.D from three independent experiments. One-way ANOVA was used for comparisons of multiple group means followed by t-test (significant compared to the stimulus only group. *p< 0.05).
$Fig. 7. Procollagen, MMP-1 and MMP-3, pro-collagen, mRNA expressions on UVB-irradiated human fibroblasts treated with Jubak fractions. The cells were treated for 24 h with the indicated concentrations of extracts before treatment with UVB (25 mJ/cm<sup>2</sup>) for 24 h. Normal : Neither extract nor UVB treatment, UVB (25 mJ/cm : only UVB treated. The data are representative of three experiments. Each value is expressed as the mean ± SD ofthee independent experiments. The statistical analysis of the data was carried out by use of ANOVA test. *p< 0.05, compared with absence of extract.$ 그림 Fig. 7. Procollagen, MMP-1 and MMP-3, pro-collagen, mRNA expressions on UVB-irradiated human fibroblasts treated with Jubak fractions. The cells were treated for 24 h with the indicated concentrations of extracts before treatment with UVB (25 mJ/cm²) for 24 h. Normal : Neither extract nor UVB treatment, UVB (25 mJ/cm : only UVB treated. The data are representative of three experiments. Each value is expressed as the mean ± SD ofthee independent experiments. The statistical analysis of the data was carried out by use of ANOVA test. *p< 0.05, compared with absence of extract.

참고문헌 (23)

J. H. Jeong, "한국인의 피부노화 특징 및 발생기전", Journal of the society of cosmetic scientists of Korea, Vol.27, No.2, pp. 31-35, (2001).？
S. H. Hong, H. N. Lee, T. K. Jeong, Y. H. Kim, H. S. Jeong, J. S. Lee, "Protective Effect of Jujube Water Extract against UVB-Induced Skin Photoaging in Human Skin Fibroblasts", Journal of the Korean Society of Food Science and Nutrition, vol. 48, No. 10, pp. 1172-1177, (2019).？
K. H. Lee, H. S. Park, l. J. Yoon, Y. B. Shin, Y. C. Baik, D. H. Koo, S. K. Kim, H. K. Jung, M. O. Sim, H. W. Cho, W. S. Jung, M. S. Kim "Whitening and Anti-Wrinkle Effects of Tremella Fuciformis Extracts", Korean Journal of Medicinal Crop Science, Vol. 24, pp. 38-46, (2016).？
Nagase, Hideaki, "Matrix metalloproteinases." Zinc Metalloproteases in Health and Disease, pp.173-224, (1996).？
S. C. Lim, "Expression of matrix metalloproteinases and its inhibitor in gastric adenocarcinoma", Cancer Research and Treatment, Vol. 33, No. 3 pp. 199-206, (2001).？

상세보기
J. S. Roh, H. J. Yeom, M. J. Oh, J. Y. Lee, "A Study on the Antioxidant and MMPs Protein Expression Inhibitive Effect of Punica granatum L. Extract and Its Stabilization with Liquid Crystal Emulsion", Journal of Life Science, Vol. 31, No. 2 pp. 164-174, (2021).？
S. Kim, T. j. Park, J. S. Lim, S. Y. Kim, "Effect of Agaricus biorenovate Extract on Collagen Synthesis and Matrix Metalloproteinase-1 Production in Human Dermal Fibroblast" KSBB Journal, Vol. 34, No. 1, pp. 49-53, (2019).？

상세보기
E. H. Lee, S. Y. Lee, H. Y. Jung, I. K. Kang, D. H. Ahn, Y. J. Cho, "Photo-aging regulation effects of newly bred Green ball apple", Journal of Applied Biological Chemistry, Vol. 63, No. 1, pp. 75-82, (2020).？
S. M. Lee, S. J. Lee, Y. Y. Kwon, S. H. Baek, J. S. Kim, H. Y. Sohn, W. C. Shin, "Skin whitening and anti-wrinkle effects of extract from Jubak of oriental herbal liquor", Journal of the Korean Society of Food Science and Nutrition, Vol. 43, No. 11, pp. 1695-1700, (2014).？
M. S. Kim, D. K. Kang, W. C. Shin, H. Y. Sohn, "Anti-microbial, anti-oxidant, and anti-thrombosis activities of the lees of Bokbunja wine (Rubus coreanus Miquel)", Journal of Life Science, Vol. 25, No. 7, pp. 757-764, (2015).？
S. M. Kim, E. H. Kim, D. H. Song, Y. S. Jung, H. J. Jeon, Y. K. Choi, H. J. Kim, "Extraction and Fermentation of Rice Wine Lees for the Development of Cosmetic Ingredients", Journal of the Korean Society of Cosmetology, Vol. 19, No. 3, pp. 501-508, (2013).？
R. L. Prior, X. Wu, K. Schaich, "Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements", Journal of agricultural and food chemistry, Vol.53, No.10, pp. 4290-4302, (2005).？

상세보기
M. J. Jang, D. H. Jun, S. H. Kim, S. I. Han, J. T. Lee, "Anti-wrinkle Effect of Chestnut Leaf", Journal of Life Science, Vol.23, No.6, pp. 751-756, (2013).？
D. H. Kim, T. S. Park, J. H. Son, "Anti-wrinkle Activities Verification of Buplerum falcatum Extracts on CCD-986sk", Journal of Applied Biological Chemistry, Vol.58, No.2, pp. 183-187, (2015).？
W. Y. Jung, J. M. Jeong, "Change of antioxidative activity at different harvest time and improvement of atopic dermatitis effects for persimmon leaf extract", The Korea Journal of Herbology, Vol. 27, No. 1, pp. 41-49, (2012).？
Y. B. Cho, J. Y. Kim, N. W. Kwon, B. Y. Hwang, J. G. Kim, S. H. Woo, M. S. Lee, "Purification and Identification of Cytotoxic Compounds from the Root of Rumex crispus L.", Korean Journal of Medicinal Crop Science, Vol. 27, No. 3, pp. 208-217, (2019).？
Park hye ryeon, & AHN HONG SEOK, "Effect of Procollagen Type I Stimulation and Inhibiton of MMP-2 mRNA Expression from Chenopodium album Ethanol Extract", Journal of the Korean Society of Cosmetology, Vol. 21, No. 6, pp. 1189-1195, (2015).？
D. H, Kim, T. S. Park, J. H. Son, "Potential prevention effects of Rubus occidentalis seed on UVB-induced MMP-1 production and procollagen degradation in CCD-986sk cells", Journal of Applied Biological Chemistry, Vol. 59, No. 4, pp. 317-322, (2016).？

원문보기 상세보기
J. E. Kim, Y. S. Choi, H. K. Kim, Y. A. Jang, "Collagen synthesis ability and inhibitory effect of MMPs in keratinocytes of Lysimachia christinae Hance Extract", The Korean Society of Applied Science and Technology, Vol. 37, No. 4, pp. 820-829, (2020).？
Y. A. Jang, "A Verification of Cosmetic Effect about Anti-oxidant and Anti-wrinkle of 11 Native Plants", Journal of Life Science, Vol. 26, No. 7, pp. 782-788, (2016).？
S. H. Lee, D. Y. Choi, Y. H. Lee, "Response to Bee Venom Acupuncture and Polymorphism of Matrix Metalloproteinase-1 Gene in Korean Patients with Rheumatoid Arthritis", Journal of Acupuncture Research, Vol .21, No. 1, pp. 211-225, (2004).？
Y. A. Jang, J. T. Lee, "Anti-wrinkle effect of berberine by inhibition of MMP-2 and MMP-9 activity in fibroblasts", Journal of Applied Biological Chemistry, Vol. 61, No. 1, pp. 9-15, (2018).？
B. A. Kim, "Anti-aging effect of Codium fragile extract on keratinocytes damaged by fine dust PM10", The Korea Jounrnal of Herbology, Vol. 38, No. 4, pp. 45-52, (2023).

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

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].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증