• 생명과학회지
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• 제28권11호
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• pp.1268-1276
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• 2018

Cytidine analog decitabine (DEC)은 핵산 합성의 억제제로서 골수이형성 증후군 및 급성 골수성 백혈병 치료제로 사용되고 있다. 산화질소 합성에서 번역 단계를 억제하는 것으로 알려진 ammonium pyrrolidine dithiocarbamate (PDTC)는 $NF-{\kappa}B$의 대표적인 억제제이다. 본 연구에서는 인체 위암 AGS 세포를 대상으로 DEC와 PDTC의 병용 처리에 따른 세포증식 억제 기전을 조사하였다. 본 연구의 결과에 따르면 PDTC에 의한 AGS 세포의 증식 억제 효과는 DEC에 의해 농도 의존적으로 유의하게 증가하였으며, 이는 G2/M기의 세포주기 정지 및 apoptosis 유도와 관련이 있었다. PDTC와 DEC의 병용 처리에 의한 세포 사멸의 유도는 DNA 손상 유도와 관련이 있음을 H2AX의 인산화 증가로 확인하였다. 아울러 PDTC와 DEC의 병용 처리는 미토콘드리아 막 전위의 파괴를 유도하고, 세포 내 활성산소종(ROS)의 생성과 Bax의 발현을 향상시키고, Bcl-2 발현을 감소시켰으며 미토콘드리아에서 세포질로의 cytochrome c 유출을 증가시켰다. 또한 PDTC과 DEC의 병용 처리는 외인성 및 내인성 apoptosis 개시 caspase에 해당하는 caspase-8과 caspase-9의 활성뿐만 아니라 caspase-3의 활성화와 PARP 단백질의 분해를 유도하였다. 결론적으로 본 연구의 결과는 PDTC와 DEC의 병용 처리가 DNA 손상을 유발하고, ROS 증가와 연계된 외인성 및 내인성 apoptosis 사멸 경로를 활성화시킴으로써 AGS 세포의 증식을 억제하였음을 의미한다.

• Biomolecules & Therapeutics
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• 제29권6호
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• pp.685-696
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• 2021

In this study, we investigated the inhibitory effect of 5-aminolevulinic acid (ALA), a heme precursor, on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages by estimating nitric oxide (NO), prostaglandin E2 (PGE2), cytokines, and reactive oxygen species (ROS). We also evaluated the molecular mechanisms through analysis of the expression of their regulatory genes, and further evaluated the anti-inflammatory and antioxidant efficacy of ALA against LPS in the zebrafish model. Our results indicated that ALA treatment significantly attenuated the LPS-induced release of pro-inflammatory mediators including NO and PGE2, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. ALA also inhibited the LPS-induced expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, reducing their extracellular secretion. Additionally, ALA abolished ROS generation, improved the mitochondrial mass, and enhanced the expression of heme oxygenase-1 (HO-1) and the activation of nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the ALA-mediated inhibition of pro-inflammatory cytokines production and activation of mitochondrial function in LPS-treated RAW 264.7 macrophages. Furthermore, ALA significantly abolished the expression of LPS-induced pro-inflammatory mediators and cytokines, and showed strong protective effects against NO and ROS production in zebrafish larvae. In conclusion, our findings suggest that ALA exerts LPS-induced anti-inflammatory and antioxidant effects by upregulating the Nrf2/HO-1 signaling pathway, and that ALA can be a potential functional agent to prevent inflammatory and oxidative damage.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.69-74
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• 2021

만성 알코올 섭취는 산화적 스트레스 및 치매와 같은 신경퇴행성질환을 발병시킨다. 본 연구는 홍화씨 추출물의 in vitro 항산화 활성과 ethanol로 손상을 유도한 신경교세포에서의 보호 효과를 확인하기 위해 수행되었다. 홍화씨 추출물은 50, 100, 250, 500 ㎍/mL의 농도에서 농도 의존적으로 in vitro에서 1,1-diphenyl-2-prcrylhydrazy (DPPH), hydroxyl radical (·OH), superoxide radical 및 nitric oxide radical 소거능이 증가하였다. 특히 DPPH 및 ·OH radical 소거능 측정 결과, 각각 500 ㎍/mL, 100 ㎍/mL의 농도에서 80% 이상의 radical 소거능을 나타내었다. 홍화씨 추출물의 신경교세포 보호 효과를 확인하기 위해, C6 신경교세포에 500 mM ethanol을 처리하여 산화적 손상을 유도하였다. Ethanol을 처리한 C6 신경교세포는 세포 생존율의 감소 및 reactive oxygen species (ROS) 생성량이 증가하여 세포 손상을 나타내었다. 반면 홍화씨 추출물을 처리하였을 때 세포 생존율 증가 및 ROS 생성 억제를 통해 ethanol로 유도된 신경교세포 손상에 대한 보호 효과를 나타내었다. 본 연구를 통해, 홍화씨는 알코올로 유도된 신경교세포의 산화적 스트레스에 대한 보호 효과가 있는 것으로 사료된다.

• 생명과학회지
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• 제32권8호
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• pp.647-658
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• 2022

미세먼지는 유해한 다양한 작은 입자를 가진 대기오염 물질로서 산화적 및 염증성 반응을 촉진하여 다양한 질환의 발병과 진행에 관여하는 것으로 알려졌다. 본 연구에서는 외부 오염물질에 직접적으로 노출되는 1차 노출 기관인 안구를 미세먼지로부터 보호할 수 있는 약재를 선정하기 위해, 인간 각막상피세포에서 후보 약물들의 방어 효능을 평가했습니다. 그 결과, 12종의 후보 약재 중 PM_2.5에 의한 세포 독성 억제효능을 보인 후보 약재 5가지(천문동, 석창포, 황련, 감국 및 금잔화)를 선별하였다. 이들 후보 물질들의 항산화 활성을 평가하기 위하여 ROS 소거능을 조사한 결과, 석창포, 천문동 및 황련 추출물이 유의한 효과를 나타내었으며, 이는 미토콘드리아 활성 보존 효능과 다소 연관성이 있었다. 또한, 이들은 PM_2.5에 의한 DNA 손상을 차단할 수 있었음을 8-OHdG 생성 및 γ-H2AX 발현 분석을 통하여 확인하였다. 본 연구의 결과는 PM_2.5에 대한 각막상피세포의 보호 신규 천연물의 탐색을 위한 자료로 활용될 것이다.

• Biomolecules & Therapeutics
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• 제32권5호
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• pp.647-657
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• 2024

Gefitinib is the well-tolerated first-line treatment of non-small cell lung cancer. As it needs analgesics during oncology treatment, particularly in the context of the coronavirus disease, where patients are more susceptible to contract high fever and sore throat. This has increased the likelihood of taking both gefitinib and antipyretic analgesic acetaminophen (APAP). Given that gefitinib and APAP overdose can predispose patients to liver injury or even acute liver failure, there is a risk of severe hepatotoxicity when these two drugs are used concomitantly. However, little is known regarding their safety at therapeutic doses. This study simulated the administration of gefitinib and APAP at clinically relevant doses in an animal model and confirmed that gefitinib in combination with APAP exhibited additional hepatotoxicity. We found that gefitinib plus APAP significantly exacerbated cell death, whereas each drug by itself had little or minor effect on hepatocyte survival. Mechanistically, combination of gefitinib and APAP induces hepatocyte death via the apoptotic pathway obviously. Reactive oxygen species (ROS) generation and DNA damage accumulation are involved in hepatocyte apoptosis. Gefitinib plus APAP also promotes the expression of Kelch-like ECH-associated protein 1 (Keap1) and downregulated the antioxidant factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), by inhibiting p62 expression. Taken together, this study revealed the potential ROS-mediated apoptosis-dependent hepatotoxicity effect of the combination of gefitinib and APAP, in which the p62/Keap1/Nrf2 signaling pathway participates and plays an important regulatory role.

• Reproductive and Developmental Biology
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• 제36권2호
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• pp.115-120
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• 2012

This study was conducted to examine the optimal concentration and treatment time of antioxidants for inhibition of the ROS generation in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine oocytes were activated parthenogenetically, during which oocytes were treated with various antioxidants to determine the optimal concentrations and kind of antioxidants. Determined antioxidants were applied to oocytes during in vitro maturation (IVM) and/or SCNT procedures. Finally, antioxidant-treated SCNT embryos were compared with in vitro fertilized (IVF) embryos. $H_2O_2$ levels were analyzed in embryos at 20 h of activation, fusion or insemination by staining of embryos in $10{\mu}M$ 2'7'-dichlorodihydrofluorescein diacetate (H2DCFDA) dye, followed by fluorescence microscopy. $H_2O_2$ levels of parthenogenetic embryos were significantly lower in $25{\mu}M$ ${\beta}$-mercaptoethanol (${\beta}$-ME), $50{\mu}M$ L-ascorbic acid (Vit. C), and $50{\mu}M$ L-glutathione (GSH) treatment groups than each control group ($24.0{\pm}1.5$ vs $39.0{\pm}1.1$, $29.7{\pm}1.0$ vs $37.0{\pm}1.2$, and $32.9{\pm}0.8$ vs $36.3{\pm}0.8$ pixels/embryo, p<0.05). There were no differences among above concentration of antioxidants in direct comparison ($33.6{\pm}0.9{\sim}35.2{\pm}1.1$ pixels/embryo). Thus, an antioxidant of $50{\mu}M$ Vit. C was selected for SCNT. $H_2O_2$ levels of bovine SCNT embryos were significantly lower in embryos treated with Vit. C during only SCNT procedure ($26.4{\pm}1.1$ pixels/embryo, p<0.05) than the treatment group during IVM ($29.9{\pm}1.1$ pixels/embryo) and non-treated control ($34.3{\pm}1.0$ pixels/embryo). Moreover, $H_2O_2$ level of SCNT embryos treated with Vit. C during SCNT procedure was similar to that of IVF embryos. These results suggest that the antioxidant treatment during SCNT procedures can reduce the ROS generation level of SCNT bovine embryos.

• 대한한의학회지
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• 제30권1호
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• pp.51-63
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• 2009

Objectives: Peroxynitrite ($ONOO^-$), superoxide anion radical (${\cdot}{O_2}^-$ and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging processes, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer, diabetes, obesity and atherosclerosis. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}{O_2}^-$ scavenging and NF-${\kappa}B$ related anti-inflammatory activities of Sotosaja-hwan in ob/ob mice. Methods: Mice were grouped and treated for 5 weeks as follows. Both the normal lean (C57/BL6J black mice) and control obese (ob/ob mice) groups have received standard chow. The experimental groups were fed with a diet of chow supplemented with 30 and 90 mg Sotosaja-hwan per 1 kg of body weight for 14 days. For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Western blotting was performed using anti-phospho-$I{\kappa}B$-${\alpha}$, anti-IKK-${\alpha}$, anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS, anti-YCAM-1 and anti-MMP-9 antibodies, respectively. Results: Sotosaja-hwan inhibited the generation of $ONOO^-$, NO and ${\cdot}{O_2}^-$ in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondrial fraction in vitro. The generation of $ONOO^-$, NO, ${\cdot}{O_2}^-$ and PGE2 were inhibited in the Sotosaja-hwan-administered ob/ob mice groups. The GSH/GSSG ratio was decreased in the ob/ob mice, whereas the ratio was improved in the Sotosaja-hwan-administered groups. Sotosaja-hwan inhibited the protein expression levels of phospho-$I{\kappa}B$-${\alpha}$, IKK-${\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, YCAM-1 and MMP-9 genes. Conclusions: These results suggest that Sotosaja-hwan is an effective $ONOO^-$, ${\cdot}{O_2}^-$ and NO scavenger and has NF-kB related anti-inflammatory activity in ob/ob mice. Therefore, Sotosaja-hwan might be a potential therapeutic drug against the inflammation process and inflammation-related diseases.

• 한국식품저장유통학회지
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• 제19권3호
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• pp.443-450
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• 2012

최근 연구에 의하면, NOX4에 의해 생성된 세포내 ROS는 지방세포의 분화를 촉진하는 것으로 알려져 있다. 본 연구진의 보고에 의하면 천년초 추출물은 $PPAR{\gamma}$의 발현 및 지방세포의 분화를 억제하는 것으로 밝혀진 바 있다. 따라서 본 연구에서는 천년초 선인장의 건강기능식품 소재화시 기초자료를 제공하고자 지방세포내 생성되는 ROS에 대한 천년초 열수 및 에탄올 추출물의 억제효과 및 주요 유전자(NOX4, G6PDH 및 항산화효소)의 발현 양상을 조사하였다. 천년초 열수 및 에탄올 추출물을 처리한 지방세포에서는 대조군에 비해 ROS 생성량이 유의적으로 감소하는 것으로 나타내었고, 추출용매별 천년초 추출물의 ROS 저감효능은 열수 추출물에 비해 에탄올 추출물에서 높게 나타났다. 천년초 추출물을 처리한 지방세포에서는 ROS 생성과 연관된 주요 유전자 NOX4 및 G6PDH의 발현이 대조군에 비해 감소하는 경향을 나타낸 반면, Cu/Zn-SOD, Mn-SOD, catalase, GPx 등과 같은 항산화효소의 발현은 대조군에 비해 증가한 것으로 나타났다. 이상의 결과들로 볼 때, 천년초 추출물은 지방세포의 분화과정에서 ROS의 생성을 억제하고 항산화효소의 발현을 증가시켜 ROS에 의한 산화적 스트레스를 효과적으로 저감화하는 것으로 밝혀졌다.

• 대한본초학회지
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• 제26권2호
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• pp.17-23
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• 2011

Objectives : This study aims at examining the effect of the fermentative extract of root of Sophorae Radix on the immuno-modulating activity. Methods : Cell viabilities were measured by MTT assay. Effect of SFS on nitric oxide(NO), hydrogen peroxide production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of SFS on productions of inflammatory cytokines such as TNF-${\alpha}$, IL-6 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Sophorae Radix. There was not any excessive toxicity to the macrophage when the fermentative extract of root of Sophorae Radix was treated in different concentrations. 2. The fermentative extract of Sophorae Radix increased the generation of hydrogen peroxide in the macrophage and significantly restored the suppression of the generation of the hydrogen peroxide in the macrophage induced by LPS. 3. The fermentative extract of Sophorae Radix reduced the generation of NO in the macrophage and significantly suppressed the increase of the generation of NO in the macrophage induced by LPS. 4. The fermentative extract of Sophorae Radix significantly decreased the amount of TNF-${\alpha}$ generated in the macrophage induced by LPS when it was $25{\mu}g/mL$ or higher. Conclusion : These results suggest that SFS has anti-inflammatory moiety related with its inhibition of NO, hydrogen peroxide, TNF-${\alpha}$, IL-6, in macrophage led by LPS.

• Preventive Nutrition and Food Science
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• 제10권1호
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• pp.46-51
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• 2005

The protective effect of Acanthopanax senticosus (AS) extract on alloxan-induced pancreatic β-cell damage was investigated in HIT T-15 cells, a Syrian hamster pancreatic β-cell line. Alloxan caused the pancreatic β-cell damage through the generation of reactive oxygen free radicals, increased DNA fragmentation, and decreased cellular NAD/sup +/ levels. The β-cell damage was significantly prevented by the pretreatment with water soluble extract of AS roots. These results suggest that the protective effect of AS extract, on alloxan-induced β-cell damage, is primarily due to the inhibition of the generation of reactive oxygen free radical species (ROS) by alloxan.