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LPS로 유도된 RAW 264.7 세포에서 청폐사간탕(淸肺瀉肝湯)의 항산화 효과
전보희,김태준,김희택,김용민,Jeon, Bo-Hee,Kim, Tae-Jun,Kim, Hee-Taek,Kim, Yong-Min 대한한방안이비인후피부과학회 2021 한방안이비인후피부과학회지 Vol.34 No.4
Objectives : This study was conducted to confirm the anti-oxidative effect of Chungpyesagan-tang(CPSGT) extract. Methods : In this study, MTT assay was performed to confirm cell viability, and DPPH and ABTS were performed to confirm radical scavenging ability. The ROS scavenging ability and the protective effect against DNA damage were confirmed by 2,7-dichlorofluorescin diacetate(DCF-DA) and 4',6-diamidino-2-phenylindole(DAPI) staining and comet assay. mRNA expression of Heme oxygenase-1(HO-1) was measured by real-time PCR, and expression of HO-1 and Kelch-like ECH-associated protein 1(Keap1) proteins was measured by western blot. Results : CPSGT was not cytotoxic at 50-400㎍/㎖. The radical scavenging activity was increased, and the ROS scavenging activity and the protective effect against DNA damage were increased compared to the LPS-treated group. The mRNA expression and protein expression of HO-1 were increased in a concentration-dependent manner. The protein expression level of Keap1 was decreased in a concentration-dependent manner. Conclusion : This suggests that CPSGT has an antioxidant effect and can be used as a potential material for skin diseases.
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전보희,송영채,김희택,김용민 대한한방안이비인후피부과학회 2023 한방안이비인후피부과학회지 Vol.36 No.1
Objectives : This study was conducted to confirm the skin anti-aging effect of Chungpyesagan-tang(CPSGT) extract. Methods : We performed MMT assay to confirm the cytotoxicity of CPSGT. After inducing matrix metalloproteinase-1(MMP-1) with tumor necrosis factor-α(TNF-α), we investigated mRNA expression and protein secretion of MMP-1 by real-time RT-PCR and ELISA. In addition, we measured the protein expression of mitogen-activated protein kinases(MAPKs) and transcription factors by western blot. Results : CPSGT was not cytotoxic at 25-800㎍/㎖. The mRNA expression and protein secretion of MMP-1 decreased when treated with CPSGT. The protein expression of p-ERK, p-JNK, p-p38 was decreased by CPSGT. In addition, the protein expression of p-c-jun and p-NF-κB, which are transcription factors, were also decreased. Conclusion : This suggests that CPSGT can inhibit MMP-1 and thus be a potential anti-aging substance.
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인간섬유아세포 Hs68에서 esculetin이 TNF-α로 유도된 MMP-1 발현에 미치는 효과
김용민,전보희 한국생약학회 2023 생약학회지 Vol.54 No.1
The skin is an important barrier that protects the body from harmful environments such as UV rays. When the skin is repeatedly stimulated, such as UV rays, ROS and pro-inflammatory cytokines are overproduced. As a result, the proteins and nucleic acids that make up the skin are damaged, and aging occurs. Esculetin is known to have anti-inflammatory, antioxidant and UV-induced MMP-1 inhibitory effects. However, the inhibitory effect of MMP-1 on TNF-α-induced fibroblasts is not known. Therefore, in this study, the MMP-1 inhibitory effect of esculetin was confirmed in TNF-α-induced fibroblasts. As a result of confirming the cytotoxicity of esculetin in Hs68 cells by MTT assay, there was no significant toxicity up to 200 μM. As a result of real-time PCR and ELISA, it was confirmed that esculetin inhibited the expression of MMP-1. Esculetin did not inhibit MAPK (ERK, JNK, p38) phosphorylation, but inhibited phosphorylation of the mTOR-p70S6k signaling pathway. In addition, it was confirmed that the phosphorylation of the transcription factor NF-κB was inhibited. These results suggest that esculetin has potential as an anti-aging material.
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대식세포에서 Nrf2/HO-1경로를 통한 청심연자음의 항산화효과
전선홍 ( Seon Hong Jeon ),오솔라 ( Sol La Oh ),김소정 ( So Jeong Kim ),전보희 ( Bo Hee Jeon ),성진영 ( Jin Young Sung ),김용민 ( Yong Min Kim ) 대한화장품학회 2020 대한화장품학회지 Vol.46 No.3
활성산소종(reactive oxygen species, ROS)은 우리 몸의 항상성 유지에 있어 중요한 역할을 한다. 그러나 과도한 ROS의 생성은 단백질, 지질, 핵산과 같은 세포 구성성분을 손상시키고 피부노화를 촉진시킨다. 이에 본 연구에서는 과도한 산화 스트레스를 예방하기 위해 Chungsimyeonja-um (CSYJE)의 항산화 효과를 확인하였다. 먼저 DPPH 및 ABTS assay를 실시하여 CSYJE의 항산화 효과를 확인한 결과 농도 의존적으로 radical 소거 활성을 확인하였다. 세포생존율 확인을 위해 MTT assay를 실시한 결과 1,000 μg/mL 농도에서 세포 독성이 없음을 확인하였다. 항산화 관련 단백질인 nuclear-E2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1)의 발현 수준을 확인하기 위해 western blotting을 실시한 결과 농도 의존적으로 발현이 증가하는 것을 확인하였다. 세포 내 ROS유발 물질인 lipopolysaccharide (LPS)로 ROS를 유도한 후, ROS생성억제효과를 확인하기 위해 DCF-DA 염색법을 실시한 결과 농도 의존적으로 ROS 생성 억제효과를 확인하였으며 ROS의 생성으로 인한 염증성 사이토카인과 염증인자의 mRNA발현 수준을 확인하기 위해 real-time RT-PCR을 실시한 결과 농도 의존적으로 염증성 사이토카인과 염증인자의 mRNA 발현을 억제시켰다. 따라서, 본 연구는 Nrf2/HO-1 신호 전달 경로 활성을 통해 CSYJE의 항산화효과를 확인했으며 이는 CSYJE가 활성산소를 억제하여 항산화 화장품의 재료로서 사용될 수 있음을 시사한다. Reactive oxygen species (ROS) plays an important role in maintaining homeostasis. However, excessive ROS production damages cellular components such as proteins, lipids, and nucleic acids and promotes skin aging. In this study, we confirmed the antioxidant effect of CSYJE to prevent excessive oxidative stress. First, DPPH and ABTS assays were performed to confirm the antioxidant effect of CSYJE and the radical scavenging activity was confirmed depending on the concentration. As a result of performing the MTT assay to confirm the cell viability, it was confirmed that there was no cytotoxicity at a concentration of 1,000 μg/mL. As a result of western blotting to confirm the expression levels of the antioxidant-related proteins nuclear-E2-related factor 2 (Nrf2) and Heme oxygenase-1 (HO-1), it was confirmed that the expression was increased in a concentration-dependent manner. After inducing ROS with lipopolysaccharide (LPS), an intracellular ROS-causing substance, DCF-DA was performed to confirm the inhibitory effect of ROS production, and the inhibition of ROS production was confirmed to concentration-dependent. Real-time RT-PCR was performed to confirm the mRNA expression level of inflammatory cytokines and inflammatory mediator caused by ROS generation, mRNA expression was reduced in a dose dependent manner. Therefore, this study confirmed the antioxidant effect of CSYJE through the Nrf2/HO-1 signaling pathway, which suggests that CSYJE can be used as an antioxidant cosmetic material by inhibiting free radicals.
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