• Journal of Life Science
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• v.28 no.12
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• pp.1406-1415
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• 2018

Based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of Houttuynia cordata Thunb, the cytoprotective effects by oxidative-stress were here analyzed. Regarding the antioxidant activity of organic solvent fractions, the electron-donating ability of DPPH increased in a dose-dependent manner, and $ED_{50}$ exhibited the highest concentration at $175{\mu}g/ml$ in the Hc-EtOAc fraction. The cell viability of Hc-EtOAc fractions on $H_2O_2$-induced HaCaT cell death ($IC_{50}$) increased in a concentration-dependent manner and a visible cell survival rate of 74% was observed at a concentration of $100{\mu}g/ml$. Meanwhile, the gene expression patterns in HaCaT cells treated with $100{\mu}g/ml$ of the Hc-EtOAc fraction for 6 and 24 hr were identified with microarray analysis. The genes involved in signal transduction, cell division, antioxidant activity, and epithelial cell proliferation were found to be 2-fold up-regulated genes in HaCaT cells following the Hc-EtOAc fraction treatment. Especially, proinflammatory cytokines (IL1B, TNF, and IL6) were identified as involved in antioxidant activity based on the expression patterns of the HaCaT cells, and pathway analysis indicated that TLR4 might be considered an upstream regulator of these genes. In order to verify the activity of IL1B, TNF, and IL6, qRT-PCR showed that the expression increased more than 2 times in HaCaT cells treated with at least $100{\mu}g/ml$ of the Hc-EtOAc fraction. The activity of the upstream regulator TLR4 protein was also increased by the Hc-EtOAc fraction. As a result, the antioxidative activity of the Hc-EtOAc fraction is predicted to pass from TLR4 through cytokines such as IL1B, TNF, and IL6.

• Journal of Bio-Environment Control
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• v.28 no.4
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• pp.366-375
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• 2019

This experiment aims to determine the proper irrigation scheduling based on a whole-substrate capacitance using a newly developed device (SCMD) by comparing with the integrated solar radiation automated irrigation system (ISR) and sap flow sensor automated irrigation system (SF) for the cultivation of tomato (Solanum lycopersicum L. 'Hoyong' 'Super Doterang') during spring to winter season. For the SCMD system, irrigation was conducted every 10 minutes after the first irrigation was started until the first run-off was occurred, of which the substrate capacitance was considered to be 100%. When the capacitance threshold (CT) was reached to the target point, irrigation was re-conducted. After that, when the target drain volume (TDV) was occurred, the irrigation stopped. The irrigation volume per event for the SCMD was set to 50, 75, or 100 mL at CT 0.9 and TDV 100 mL during the spring to summer cultivation, and the CT was set to 0.65, 0.75, 0.80, or 0.90 in the winter cultivation. When the irrigation volume per event was set to 50, 75, or 100 mL, the irrigation frequency in a day was 39, 29, and 19, respectively, and the drain rate was 3.04, 9.25, and 20.18%, respectively. When the CT was set to 0.65, 0.75, or 0.90 in winter, the irrigation frequency was about 6, 7, 15 times, respectively and the drain rate was 9.9, 10.8, 35.3% respectively. The signal of stem sap flow at the beginning of irrigation starting time did not correspond to that of solar irradiance when the irrigation volume per event was set to 50 or 75 mL, compared to that of 100 mL. In winter cultivation, the stem sap flow rate and substrate volumetric water content at the CT 0.65 treatment were very low, while they were very high at CT 0.90 was high. All the integrated data suggest that the proper range of irrigation volume per event is from 75 to 100 mL under at CT 0.9 and TDV 100 mL during the spring to summer cultivation, and the proper CT seems to be higher than 0.75 and lower than 0.90 under at 75 mL of the irrigation volume per event and TDV 70 mL during the winter cultivation. It is going to be necessary to investigate the relationship between capacitance value and substrate volumetric water content by determining the correction coefficient.

• Journal of Life Science
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• v.32 no.11
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• pp.833-840
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• 2022

Chronic inflammation has been shown to be closely associated with tumor development and progression. Nuclear factor kappa B (NF-κB) is composed of a family of five transcription factors. NF-κB signaling plays a crucial role in the inflammatory response and is often found to be dysregulated in various types of cancer, making it an attractive target in cancer therapeutics. In this study, CDC-like kinase 3 (CLK3) was identified as a novel kinase that regulates the NF-κB signaling pathway. Our data demonstrate that CLK3 inhibits the canonical and non-canonical NF-κB pathways. Luciferase assays following the transient or stable expression of CLK3 indicated that this kinase inhibited NF-κB activation mediated by Tumor necrosis factor-alpha (TNFα) and Phorbol 12-myristate 13-acetate (PMA), which are known to activate NF-κB signaling via the canonical pathway. Consistent with data on the ectopic expression of CLK3, CLK3 knockdown using shRNA constructs increased NF-κB activity 1.5-fold upon stimulation with TNFα in HEK293 cells compared with the control cells. Additionally, overexpression of CLK3 suppressed the activation of this signaling pathway induced by NF-κB-inducing kinase (NIK) or CD40, which are well-established activators of the non-canonical pathway. To further examine the negative impact of CLK3 on NF-κB signaling, we performed Western blotting following the TNFα treatment to directly identify the molecular components of the NF-κB pathway that are affected by this kinase. Our results revealed that CLK3 mitigated the phosphorylation/activation of transforming growth factor-α-activated kinase 1 (TAK1), inhibitor of NF-κB kinase alpha/beta (IKKα/α), NF-κB p65 (RelA), NF-κB inhibitor alpha (IκBα), and Extracellular signal-regulated kinase 1/2-Mitogen-activated protein kinase (ERK1/2-MAPK), suggesting that CLK3 inhibits both the NF-κB and MAPK signaling activated by TNFα exposure. Further studies are required to elucidate the mechanism by which CLK3 inhibits the canonical and non-canonical NF-κB pathways. Collectively, these findings reveal CLK3 as a novel negative regulator of NF-κB signaling.

• Journal of Life Science
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• v.32 no.12
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• pp.929-937
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• 2022

To investigate the anti-inflammatory activity of the grains of sorghum, three Sorghum bicolor (L.) Moench variants (Hwanggeumchal, Huinchal, and Chal) being cultivated in Korea, the 80% ethanol (EtOH) extracts of individual sorghum grains were compared for their inhibitory activity against nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cell line. Among them, the EtOH extract of sorghum Hwanggeumchal grains could exert the highest inhibitory effect on the LPS-induced NO production. However, under these conditions, the viability of RAW264.7 cells was not affected. When the EtOH extract of sorghum Hwanggeumchal grains was sequentially fractionated with n-hexane, methylene chloride (MC), ethyl acetate (EtOAc), and n-butanol, the anti-NO production activity was predominantly detected in both MC and EtOAc fractions. In particular, treatment with the MC fraction reduced dose-dependently the expression levels of iNOS, COX-2 and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in LPS-stimulated RAW264.7 cells. Simultaneously, the MC fraction could prevent LPS-induced activating phosphorylation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). HPLC analysis of the MC fraction showed gentisic acid and naringenin as the major phenolic components. Both gentisic acid and naringenin commonly exhibited a potent inhibitory activity against LPS-induced NO production in RAW264.7 cells. Together, these results provide the evidence of the inhibitory activity of Hwanggeumchal grains on LPS-induce inflammatory responses in RAW264.7 murine macrophage cells and also suggest that sorghum grains possess beneficial health effects which can be applicable in development of the grain-based functional foods.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.204-212
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• 2023

Whitening is inhibitory activity of the melanin synthesis of melanocytes. Recently, whitening materials have been developed on natural materials because of its side effects on skin. Figs (Ficus Carica L.) is a fruit belonging to the Moraceae family and whitening activity was reported in focusing on the fig's stem and leaf components, but whitening activity of the figs fruit was not known. Thus, in this study, we tried to observe its anti-melanogenesis as well as antioxidant and anti-inflammation. The radical scavenging activity of figs fruits extract (FFE) was observed as the level of 34.52±1.98%/60.71±1.26% compared to the control in the its maximum concentration in the DPPH/ABTS assay. Cytotoxicity of FFE was observed at 10% concentration by CCK8 assay, so the maximum concentration was set at 5% and applied to all experiments. FFE concentration dependently decreased NO production associated with inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-α gene expression, these strongly suggesting anti-inflammatory activity. In melanin contents assay, FFE significantly down-regulated melanin production in α-MSH-stimulated B16F10 cell as well as tyrosinase inhibition in vitro. In addition, FFE decreased the Microphthalmia-associated transcription factor (MITF) mRNA expression about 94.34% compared to the α-MSH treatment group in RT-PCR. Finally, FFE significantly reduced the MITF, cAMP response element-binding protein and tyrosinase protein expression in the α-MSH stimulated B16F10 cell. Through these results, we found that FFE can not only directly inhibit tyrosinase enzyme activity but also suppress melanogenesis through regulation of MITF gene expression in α-MSH signal transduction.