[논문]연교의 항산화 효과 연구

김영은; 김민진; 배수진; 박선빈; 박선동; 박광일; 김영우

doi:10.14374/hfs.2024.32.1.51

연교의 항산화 효과 연구
Anti-oxidant effect of forsythia suspensa on cellular damage in the chronic disease 원문보기

大韓韓醫學方劑學會誌 = Herbal formula science, v.32 no.1, 2024년, pp.51 - 61

김영은 (동국대학교 한의과대학 방제학교실) , 김민진 (동국대학교 한의과대학 방제학교실) , 배수진 (동국대학교 한의과대학 방제학교실) , 박선빈 (동국대학교 한의과대학 방제학교실) , 박선동 (동국대학교 한의과대학 방제학교실) , 박광일 (경상대학교 수의과대학 생리학 실험실) , 김영우 (동국대학교 한의과대학 방제학교실)

Abstract ▼ AI-Helper

Objectives : This study induced oxidative stress in HepG2 cells by treating them with AA+iron and investigated the effects of forsythia suspensa extract on this stress, as well as elucidated the molecular mechanisms underlying its hepatoprotective effects. Methods : To confirm the antioxidative effects of FSE, HepG2 cells were induced with AA+iron to induce oxidative stress, followed by MTT assay. Additionally, the effect of FSE in reducing the increased ROS levels and mitochondrial damage induced by AA+iron in HepG2 cells was confirmed using FACS. Furthermore, western blot analysis were conducted to investigate the molecular mechanisms underlying the hepatoprotective effects of FSE. Results : FSE increased the decreased cell viability induced by AA+iron. Additionally, FSE normalized the expression of apoptosis-related proteins induced by AA+iron. The elevated ROS levels in HepG2 cells induced by AA+iron were reduced by FSE, and the increase in Rh123-negative cells induced by AA+iron was attenuated by FSE. Moreover, FSE activated the protein expression of AMPK and its related phosphorylating enzymes, LKB1 and ACC. Furthermore, FSE activated YAP and its upstream phosphorylating enzyme, LATS1. Conclusions : These results demonstrate that FSE has an inhibitory effect on oxidative stress induced by AA+iron and may have potential hepatoprotective effects.

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표/그림 (4)

그림 Fig 1. Effect of FSE on cytotoxicity. (A)　Cell viability were measured through the MTT assay. HepG2 cells were induced with AA+iron and treated with FSE (1, 3, 10, 30 μg/mL). The data were presented as the mean ± SD from three independent experiments (**p < 0.01 vs. control group; #p <0.05 vs. AA+iron-treated group). (B)　Western blot analysis was performed to examine the expression of apoptosis-related proteins. HepG2 cells were treated with AA and FSE (30 μg/ml) for 12h, followed by a 1h iron treatment.
그림 Fig 2. Effect of FSE on ROS generation and mitochondrial dysfunction. (A) The level of intracellular ROS generation was assessed using the DCF-DA assay. HepG2 cells were treated with AA and FSE (30 μg/mL) for 12 h, followed by a 2 h iron treatment. Subsequently, the cells were stained with DCF-DA (30 μM) for 1 h. (B) Mitochondrial dysfunction within the cells was measured by staining with Rh123. HepG2 cells were treated with AA and FSE (30 μg/mL) for 12 h, followed by a 2 h iron treatment. The data were presented as the mean ± SD from three independent experiments (**p < 0.01 vs. control group; ##p < 0.01 vs. AA+iron-treated group).
그림 Fig 3. Effects of FSE on AMPK activation. (A) Western blotting for the AMPK signaling pathway. HepG2 cells were treated with FSE (30 μg/ml) at different time points (10', 30', 1h, 3h, 6h). The data are presented as the mean ± SD from three independent experiments (*p < 0.05, **p < 0.01 vs. the control group). (B) Huh7 cells were treated with FSE (30 μg/ml) at different time points (10', 30', 1h, 3h, 6h).
그림 Fig 4. Effects of FSE on YAP activation. (A) Western blotting for the YAP signaling pathway. HepG2 cells were treated with FSE (30 μg/ml) at different time points (10', 30', 1h, 3h, 6h). The data are presented as the mean ± SD from three independent experiments (*p < 0.05, **p < 0.01 vs. the control group). (B) Huh7 cells were treated with FSE (30 μg/ml) at different time points (10', 30', 1h, 3h, 6h).

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