• 대한한의학회지
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• 제30권6호
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• pp.17-26
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• 2009

Objectives: Little information is available regarding the effect of Lycii cortex radicis (LCR) on cell viability in glioma cells. This study was therefore undertaken to examine the effect of LCR on cell survival in U87MG human glioma cells. Methods: Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. Activation of Akt and extracellular signal-regulated kinase (ERK) and activation of caspase-3 were estimated by Western blot analysis. Results: LCR resulted in apoptotic cell death in a dose- and time-dependent manner. LCR increased reactive oxygen species (ROS) generation and LCR-induced cell death was also prevented by antioxidants, suggesting that ROS generation played a critical role in LCR-induced cell death. Western blot analysis showed that LCR treatment caused down-regulation of Akt and ERK. The LCR-induced cell death was increased by the inhibitors of Akt and ERK. Activation of caspase-3 was stimulated by LCR and caspase inhibitors prevented the LCR-induced cell death. Conclusion: These findings suggest that LCR results in human glioma cell death through a mechanism involving ROS generation, down-regulation of Akt and ERK, and caspase activation.

• Archives of Pharmacal Research
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• 제27권2호
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• pp.194-198
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• 2004

Reactive oxygen species (ROS) play an important role in the pathogenesis of many human degenerative diseases such as cancer, aging, arteriosclerosis, and rheumatism. Much attention has been focused on the development of safe and effective antioxidants. To discover sources of antioxidative activity in marine algae, extracts from 17 kinds of seaweed were screened for their inhibitory effect on total ROS generation in kidney homogenate using 2',7'-dichlorofluorescein diacetate (DCFH-DA). ROS inhibition was seen in three species: UIva pertusa, Symphyocladia latiuscula, and Ecklonia stolonifera. At a final concentration of 25 $\mu\textrm{g}$/mL, U. pertusa inhibited 85.65$\pm$20.28％ of total ROS generation, S. latiscula caused 50.63$\pm$0.09％ inhibitory, and the Ecklonia species was 44.30$\pm$7.33％ inhibition. E. stolonifera OKAMURA (Lam-inariaceae), which belongs to the brown algae, has been further investigated because it is commonly used as a foodstuff in Korea. Five compounds, phloroglucinol (1), eckstolonol (2), eckol (3), phlorofucofuroeckol A (4), and dieckol (5), isolated from the ethyl acetate soluble fraction of the methanolic extrclct of E. stolonifera inhibited total ROS generation.

• 동의생리병리학회지
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• 제29권6호
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• pp.475-484
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• 2015

In this paper, we studied the anti-oxidative capacities and protective effects of water extract of Gagam-Danguieumja(GDE) against Ultraviolet B(UVB)-induced oxidative damage in human keratinocytes(HaCaT). To evaluate the anti-oxidative activities of GDE, we measured scavenging activities on DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation and reducing power of GDE. To detect the protective effects of GDE against UVB, we irradiated with 40 mJ/㎠`s UVB to HaCaT cells then we measured reactive oxygen species(ROS) generation, apoptotic bodies and cell viability using DCFH-DA assay, Hoechst 33342 staining and MTT assay. GDE showed the anti-oxidative activities by scavenging DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation. Also GDE showed high reducing values. GDE reduced oxidative stress conditions by inhibition of ROS expression. Also the cell apoptosis by UVB-induced oxidative conditions was decreased by GDE treatment. These results could suggest that GDE had anti-oxidative activities and exhibited protective effects against UVB on HaCaT cells. GDE would be useful for the development of cosmetics treating UVB-induced skin aging.

• 한방재활의학과학회지
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• 제26권3호
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• pp.51-58
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• 2016

Objectives This study was designed to investigate whether the Cheongajihwang-Tang (CT) has an inhibitory effect association with oxidation or inflammation in RAW264.7 cells. Methods Cytotoxic activity of CT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Nitric oxide production was measured using Griess reagent system. The total phenolic contents and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the anti-oxidative effects of CT. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production. Results Cheongajihwang-Tang does not impair the cell viability in tested concentration. CT showed anti-oxidative effects in vitro by decreasing electron donating ability, and also showed anti-inflammatory effects suppressing NO and ROS expression in LPS induced RAW264.7 activation. CT inhibited the generation of intracellular ROS production as dose dependant manner. Conclusions CT has anti-oxidative effects and anti-inflammatory activities. These results indicate that CT extract has an anti-inflammatory activities via anti-oxidative effects.

• Korean Journal of Acupuncture
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• 제25권4호
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• pp.89-104
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• 2008

Objectives : This study was undertaken to determine the in vitro antioxidant activity of the extract of Chilgitang herb-acupuncture solution (CHAS). Methods : The radical scavenging capacity was tested by 2,2-diphenyl-1-picryl-hydrazyl (DPPH), hypoxanthine-xanthine oxidase system, DCFH-DA assay, nitric oxide and peroxynitrite generating system. In addition, antioxidant activity on copper and AAPH mediated human low-density lipoprotein (LDL) oxidation was measured by using TBARS assay and relative electrophoretic mobility assay. The amount of total phenolic compounds was assayed by the Folin-Ciocalteu method. Results : CHAS revealed a potent scavenging activity on DPPH radical(82%), superoxide anions(73%), hydroxyl radical(63%), nitric oxide (99%) and peroxynitrite (99%). Moreover, CHAS showed a strong inhibitory effect (59%) on $FeCl_2$-ascorbic acid induced lipid peroxidation of rat liver homogenate. CHAS also markedly inhibited copper(81%) and AAPH(56%)-mediated LDL oxidation, and effectively suppressed the electrophoretic mobility during exposure of human LDL to copper ions. CHAS (82 mg/g) contained higher concentration of total phenolic compounds than that of water extract (45 mg/g) obtained from Chilgitang. Conclusions : These results indicate that CHAS may protect against ROS- or RNS involved diseases, including cardiovascular diseases.

• 통합자연과학논문집
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• 제15권3호
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• pp.123-129
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• 2022

Schwann cells play a critical role for myelination in peripheral nerve system. It also plays an important role in nerve protection and regeneration. In peripheral nerve damage, regeneration is induced by the migration and proliferation of Schwann cells which were promoted by suppressing the oxidative stress. In this study, Human placental extract was prepared by homogenization and estimated its efficacy in RSC96 cells. Placental extract exhibited a protective effect against hydrogen peroxide-induced oxidative stress in RSC96 cells, confirmed by MTT assay. Furthermore, placental extract decreased intracellular ROS against oxidative stress, confirmed by DCFH-DA assay. Autophagy was visualized with Cyto-ID staining to confirm the autophagy activity of placental extracts. The activity of autophagy was confirmed by immunoblot analysis of autophagy flux-associated proteins such as LC3 conversion and SQSTM1 degradation. Thus, we confirmed the antioxidant effect of placental extract to protect RSC96 cells from oxidative stress, and observed that it activated autophagy and restored autophagy flux.

• Korean Journal of Acupuncture
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• 제29권3호
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• pp.406-420
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• 2012

Objectives : Scutellaria barbata has been widely used in oriental medicine used for treatment of acute and chronic inflammatory diseases. In this study, to investigate the protective effect of Scutellaria barbata on type I allergic response, we determined whether Scutellaria barbata inhibits early or late allergic responses. Methods : To assess the effect of Scutellaria barbata Pharmacopuncture Extract(SB) in RBL-2H3 cells, we investigated the levels of the markers of degranulation such as ${\beta}$-hexosaminidase and histamine, inflammatory mediator such as IL-4, TNF-${\alpha}$, PGE2 and cysLT, and mRNA expression of cytokines and enzymes. In addition, we determined the levels of intracellular ROS by DCFH-DA assay and the free radical scavenging activity by DPPH method. Results : We found that SB suppressed the release of ${\beta}$-hexosaminidase and histamine and the production of IL-4, TNF-${\alpha}$, PGE2 and cysLT in RBL-2H3 by the antigen stimulation. SB also significantly inhibited the enzyme mRNA expressions, such as HDC2, COX-1, COX-2, 5-LOX and iNOS2, along with reduced cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, SB suppressed the levels of intracellular ROS. Conclusions : Our results indicate that SB protects against type I allergic response and exert an anti-inflammatory effect through the inhibition of degranulation, inflammatory mediator release and mRNA expression of cytokines and enzymes.

• 대한한의학방제학회지
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• 제22권1호
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• pp.79-92
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• 2014

Objectives : The aim of this study was identification of the anti-oxidative and anti-inflammatory effects of Talmyung-san (TMS) in mouse macrophage, RAW264.7 cells. Methods : To identify the anti-oxidative effect of TMS, scavenging activities of DPPH radical, nitric oxide and peroxynitrite were measured in vitro. In RAW264.7 cells, DCFH-DA assay was conducted to examine the inhibitory effect of TMS on ROS production in response to lipopolysaccharide. And the productions of nitric oxide (NO), $PGE_2$ and pro-inflammatory cytokines were measured. The levels of COX-2, iNOS, nuclear NF-${\kappa}B$ p65 expression and phosphorylation of $I{\kappa}B-{\alpha}$ in cytosol were detected by western blotting analyses. Results : TMS couldn't scavenged DPPH radical, but nitric oxide and peroxynitrite were decreased. TMS decreased intracellular ROS, NO, and IL-$1{\beta}$ production effectively. However, TMS inhibited $PGE_2$ levels only in high concentration ($300{\mu}g/m{\ell}$) and TMS failed to suppress the production of IL-6 and TNF-${\alpha}$. Results from immunoblot analyses revealed that TMS decreased activation of COX-2, iNOS, phosphorylation of $I{\kappa}B-{\alpha}$ and nuclear translocation of p65. Conclusions : TMS has anti-RNS and anti-inflammatory effects via NF-${\kappa}B$ pathway and more intensive studies will be required to evaluate therapeutic potential of TMS.

• 한국작물학회지
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• 제56권4호
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• pp.349-360
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• 2011

The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.

• The Korean Journal of Physiology and Pharmacology
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• 제24권3호
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• pp.223-232
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• 2020

Sesamin, a lipid-soluble lignin originally isolated from sesame seeds, which induces cancer cell apoptosis and autophagy. In the present study, has been reported that sesamin induces apoptosis via several pathways in human lung cancer cells. However, whether mitophagy is involved in sesamin induced lung cancer cell apotosis remains unclear. This study, the anticancer activity of sesamin in lung cancer was studied by reactive oxygen species (ROS) and mitophagy. A549 cells were treated with sesamin, and cell viability, migration ability, and cell cycle were assessed using the CCK8 assay, scratch-wound test, and flow cytometry, respectively. ROS levels, mitochondrial membrane potential, and apoptosis were examined by flow cytometric detection of DCFH-DA fluorescence and by using JC-1 and TUNEL assays. The results indicated that sesamin treatment inhibited the cell viability and migration ability of A549 cells and induced G₀/G₁ phase arrest. Furthermore, sesamin induced an increase in ROS levels, a reduction in mitochondrial membrane potential, and apoptosis accompanied by an increase in cleaved caspase-3 and cleaved caspase-9. Additionally, sesamin triggered mitophagy and increased the expression of PINK1 and translocation of Parkin from the cytoplasm to the mitochondria. However, the antioxidant N-acetyl-L-cysteine clearly reduced the oxidative stress and mitophagy induced by sesamin. Furthermore, we found that cyclosporine A (an inhibitor of mitophagy) decreased the inhibitory effect of sesamin on A549 cell viability. Collectively, our data indicate that sesamin exerts lethal effects on lung cancer cells through the induction of ROS-mediated mitophagy and mitochondrial apoptosis.