[논문]으름덩굴 에탄올 추출물의 항노화 효과

김유진; 권순현; 송지현; 이소미; 김용민

doi:10.15230/scsk.2024.50.1.67

[국내논문] 으름덩굴 에탄올 추출물의 항노화 효과
Anti-aging Effect of Akebia quinata Decaisne Ethanol Extract

大韓化粧品學會誌 = Journal of the society of cosmetic scientists of Korea, v.50 no.1, 2024년, pp.67 - 75

김유진 (세명대학교 화장품과학과) , 권순현 (세명대학교 화장품과학과) , 송지현 (세명대학교 화장품과학과) , 이소미 (세명대학교 화장품과학과) , 김용민 (세명대학교 화장품과학과)

초록
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피부는 자외선, 감염 등과 같은 외부 요인에 의해 피부 노화가 진행된다. 이러한 요인들에 의해 피부의 섬유아세포는 단백질 분해효소인 matrix metalloproteinases (MMPs)를 분비한다. MMPs는 세포외기질에 위치하는 콜라겐의 분해를 유도하여 노화에 직접적인 영향을 미친다. 으름덩굴(Akebia quinata Decaisne) 줄기는 항산화, 항염증 효과가 있는 것으로 보고되었다. 하지만 으름덩굴 줄기 에탄올 추출물(AQSEE)의 항노화 효과에 대해서는 알려지지 않았다. 따라서 인간 섬유아세포에서 TNF-α로 유도된 MMP-1 억제 효과를 연구하였다. MTT asaay를 통해 AQSEE의 세포 생존율을 확인한 결과 400 ㎍/mL까지 독성을 나타내지 않았다. RT-qPCR과 ELISA를 통해 MMP-1 mRNA와 단백질 분비를 억제하는지 확인한 결과 100, 200, 400 ㎍/mL 농도에서 유의하게 감소하였다. 또한, western blotting을 통해 MAPKs 신호전달경로와 전사인자의 인산화가 감소하는지 확인하였다. 그 결과 p38, c-Jun, p65의 인산화가 모든 농도에서 유의하게 감소하였다. AQSEE의 radical 소거능을 확인하기 위해 DPPH, ABTS assay를 진행한 결과 모든 농도에서 유의하게 감소하였다. 본 연구 결과를 통해 MMP-1 억제 효과와 radical 소거능을 확인하였으며, 이는 항노화 물질로 사용될 수 있음을 시사한다.

Abstract ▼ AI-Helper

Skin aging progresses due to external factors such as ultraviolet rays and infections. These factors cause skin fibroblasts to secrete proteolytic enzymes, matrix metalloproteinases (MMPs). MMPs induce the degradation of collagen located in the extracellular matrix, directly influencing aging. The stems of Akebia quinata Decaisne have been reported to have antioxidant and anti-inflammatory effects. However, the anti-aging effect of Akebia quinata Decaisne stem ethanol extract (AQSEE) is not known. Therefore, we studied the TNF-α-induced MMP-1 inhibitory effect in human fibroblasts. When the cell viability of AQSEE was confirmed through MTT asaay, it showed no toxicity up to 400 ㎍/mL. The inhibition of MMP-1 mRNA and protein secretion was confirmed through RT-qPCR and ELISA, and results showed a significant decrease at concentrations of 100, 200, 400 ㎍/mL. We also confirmed by Western blotting that phosphorylation of MAPKs signaling pathway and transcription factors was reduced. As a result, phosphorylation of p38, c-Jun, p65 was significantly decreased at all concentrations. DPPH and ABTS assays were performed to confirm the radical scavenging ability of AQSEE, and the results showed a significant decrease at all concentrations. The results of this study confirmed the MMP-1 inhibitory effect and radical scavenging ability, which suggests that it can be used as an anti-aging substance.

Keyword

표/그림 (9)

표 Table 1. Gene Name and Assay ID Number in RT-qPCR Analysis
표 Table 2. Antibody Name and Assay ID Number in Western Blotting Analysis
그림 Figure 1. The effects of AQSEE on cell viability in Hs68 cells. There was no cytotoxicity at 50 ~ 400 μg/mL concentrations. Cell viability was measured by MTT assay. The data represent the mean ± SD of triplicate experiments.
그림 Figure 2. The e ffects o f AQSEE on MMP -1 mRNA expression and protein secretion in TNF-α induced Hs68 cells. (A) AQSEE inhibited TNF-α induced MMP-1 mRNA expression. (B) AQSEE inhibited TNF-α induced MMP-1 protein secretion. Forskolin was positive control (10 μM). The data represent the mean ± SD of triplicate experiments. Indicates a significant difference from the control (^#p < 0.05). Indicates a significant difference from the TNF-α treated condition (*p < 0.05).
그림 Figure 3. The effects of AQSEE on MAPKs protein expression in Hs68 cells. (A) AQSEE inhibited the phosphorylation of p38. (B) p-ERK, p-38, p-JNK protein levels were compared to ERK, p38, JNK, respectively, and analyzed using Image J. Forskolin was positive control (10 μM). The data represent the mean ± SD of triplicate experiments. Indicates a significant difference from the control (^#p < 0.05). Indicates a significant difference from the TNF-α treated condition (*p < 0.05).
그림 Figure 4. The effects of AQSEE on p-c-jun protein expression in Hs68 cells. (A) AQSEE inhibited the phosphorylation of c-jun. (B) p-c-Jun protein levels were compared to c-Jun and analyzed using Image J. Forskolin was positive control (10 μM). The data represent the mean ± SD of triplicate experiments. Indicates a significant difference from the control (^#p < 0.05). Indicates a significant difference from the TNF-α treated condition (*p < 0.05).
그림 Figure 5. The effects of AQSEE on p-p65 protein expression in Hs68 cells. (A) AQSEE inhibited the phosphorylation of p65. (B) p-p65 protein levels were compared to p65 and analyzed using Image J. Forskolin was positive control (10 μM). The data represent the mean ± SD of triplicate experiments. Indicates a significant difference from the control (^#p < 0.05). Indicates a significant difference from the TNF-α treated condition (*p < 0.05).
그림 Figure 6. The effects of AQSEE on DPPH/ABTS radical scavening test. Ascorbic acid was positive control (200 μM). The data represent the mean ± SD of triplicate experiments. Indicates a significant difference from the control (*p < 0.05).
그림 Figure 7. AQSEE can suppress MMP -1 mRNA and protein secretion by inhibiting the p38 pathway.

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