[논문]염생식물인 갯끈풀과 갯메꽃 추출물의 HaCaT 세포에서 피부 보습 및 피부 장벽 기능에 미치는 영향

하유나; 정재우; 이원휘; 오준혁; 김연정

doi:10.23005/ksmls.2021.6.2.58

염생식물인 갯끈풀과 갯메꽃 추출물의 HaCaT 세포에서 피부 보습 및 피부 장벽 기능에 미치는 영향
Effect of Halophyte (Spartina anglica and Calystegia soldanella) Extracts on Skin Moisturizing and Barrier Function in HaCaT Cells 원문보기

한국해양생명과학회지 = Journal of marine life science, v.6 no.2, 2021년, pp.58 - 65

하유나 (인천대학교 기초과학연구소) , 정재우 , 이원휘 (인천대학교 기초과학연구소) , 오준혁 (인천대학교 해양학과) , 김연정 (인천대학교 기초과학연구소)

초록
AI-Helper

노화가 진행될수록 활성산소종으로 인하여 피부 보습은 떨어지고 피부 장벽은 붕괴되어 피부가 손상된다. 본 연구에서는 인천 동막 해변에 서식하는 염생식물인 갯끈풀(Spartina anglica; SAE)과 갯메꽃(Calystegia soldanella; CSE)을 70% 에탄올(EtOH)로 추출하여 피부 보습 및 피부 장벽 기능 강화에 대한 효능을 평가하였다. 이 추출물들에 대한 피부 각질형성세포(HaCaT cell)에서 세포독성을 WST-8 assay를 이용하여, 세포 생존율이 90% 이상을 보이는 농도를 선별하여 추가 실험을 진행하였다. ABTS 라디칼 소거능을 통해 항산화 효과를 확인한 결과, SAE와 CSE는 높은 라디칼 소거능을 보였다. 피부 보습과 관련된 인자들인 filaggrin (FGL), aquaporin 3(AQP3), hyaluronan synthase 2 (HAS2)과 피부 장벽 기능과 연관 있는 transglutaminase 1 (TGM1)과 involucrin (INV)의 유전자 수준에서의 발현 변화를 측정한 결과, SAE에 의해 AQP3, HAS2, TGM1의 발현이 증가하였으나, CSE는 변화가 없는 것을 확인할 수 있었다. SAE에 의한 세포 내 신호전달 경로를 확인하기 위해 western blot 분석을 수행하였다. Extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen activated protein kinase의 활성이 SAE에 의하여 상향 조절되었음을 확인하였다. 이러한 결과는 갯끈풀 추출물이 피부 보습 및 피부 장벽 기능 강화를 위한 화장품의 기능성 소재로 사용될 수 있음을 시사한다.

Abstract ▼ AI-Helper

As aging progresses, reactive oxygen species (ROS) reduces skin moisturization and collapses skin barrier function. In this study, we evaluated the efficacy of skin moisturizing and skin barrier function enhancement by extracts from halophytes using HaCaT cells. Spartina anglica (S. anglica; SAE) and Calystegia soldanella (C. soldanella; CSE), a kind of halophytes, were collected from Dongmak beach in Incheon, and extracted with 70% ethanol. At the first, we evaluated the cytotoxicity of extracts in HaCaT cell using WST-8 Kit. As a result, the other experiment was conducted by setting the concentration at which the cell viability was 90% or more. SAE and CSE showed high radical scavenging activity through ABTS assay. Expression levels of genes related to skin moisturizing and skin barrier functions, were analyzed by real-time qPCR. As a result, it showed that the expression of aquaporin 3, hyaluronan synthase 2, and transglutaminase 1 was increased by SAE treatment but not changed by CSE. Activation of extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen activated protein kinase was induced by SAE. These results suggest that SAE can be used as functional materials for cosmetics for skin moisturizing and barrier function enhancement.

주제어

표/그림 (6)

표 Table 1. Sequences of the specific target genes
그림 Fig. 1. ABTS radical scavenging activity of halophyte extracts. Free radical scavenging activities of S. anglica (SAE) (A) and C. soldanella (CSE) (B) determined by ABTS assay. L-ascorbic acid was used as a positive control. Data are expressed as percentage of control. The results were expressed as mean ± S.D. (n=3). ***p<0.001, **p<0.01 compared with control.
그림 Fig. 2. Cell viability in various treatment concentration of D-panthenol and halophyte extracts (SAE and CSE) on HaCaT cells. Cell were treated with various concentration of D-panthenol (A), S. angilca (SAE) (B) and C. soldanella (CSE) (C) for 24 h. Cell viability was measured by WST-8 cell viability assay. Results are the means ± S.D. of three independent experiments. *p<0.05, ***p<0.001 indicate a significant difference compared to the control.
그림 Fig. 3. Skin moisturization effects of halophyte extracts on mRNA expression level in HaCaT cell. Cells were treated with various concentration of the halophyte extracts S. angilca (SAE)(A) and C. soldanella (CSE) (B) for 24 h. mRNA levels of FLG, AQP3, and HAS2 genes were measured using real-time qPCR. GAPDH was used as an internal control. Each bar represents the mean ± S.D. (n=3). *p<0.05, **p<0.01, and ***p<0.001 compared with control.
그림 Fig. 4. Skin barrier function effects of halophyte extracts on mRNA expression level in HaCaT cell. Cells were treated with various concentration of the halophyte extracts S. angilca (SAE) (A) and C. soldanella (CSE) (B) for 24 h. mRNA level of INV and TGM1 were measured using real-time qPCR. GAPDH was used as an internal control. Each bar represents the mean ± S.D. (n=3). *p<0.05, **p<0.01, and ***p<0.001 compared with control.
그림 Fig. 5. Effect of halophyte extracts on the signaling pathway of MAPKs in HaCaT cells. Cells were treated varying concentrations of SAE and D-panthenol with for 24 h. Phosphorylation levels of extracellular signal-related kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 was shown in graph. The band densities were expressed as percentage of SAE or D-panthenol treated group related to control band using Image Lab analyze program. Data are presented as the means ± S.D. of independent experiments (n=3). *p<0.05, **p<0.01, and ***p<0.001 compared with control.

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