• BMB Reports
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• v.56 no.3
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• pp.178-183
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• 2023

Huntington's disease (HD) is a neurodegenerative disorder, of which pathogenesis is caused by a polyglutamine expansion in the amino-terminus of huntingtin gene that resulted in the aggregation of mutant HTT proteins. HD is characterized by progressive motor dysfunction, cognitive impairment and neuropsychiatric disturbances. Histone deacetylase 6 (HDAC6), a microtubule-associated deacetylase, has been shown to induce transport- and release-defect phenotypes in HD models, whilst treatment with HDAC6 inhibitors ameliorates the phenotypic effects of HD by increasing the levels of α-tubulin acetylation, as well as decreasing the accumulation of mutant huntingtin (mHTT) aggregates, suggesting HDAC6 inhibitor as a HD therapeutics. In this study, we employed in vitro neural stem cell (NSC) model and in vivo YAC128 transgenic (TG) mouse model of HD to test the effect of a novel HDAC6 selective inhibitor, CKD-504, developed by Chong Kun Dang (CKD Pharmaceutical Corp., Korea). We found that treatment of CKD-504 increased tubulin acetylation, microtubule stabilization, axonal transport, and the decrease of mutant huntingtin protein in vitro. From in vivo study, we observed CKD-504 improved the pathology of Huntington's disease: alleviated behavioral deficits, increased axonal transport and number of neurons, restored synaptic function in corticostriatal (CS) circuit, reduced mHTT accumulation, inflammation and tau hyperphosphorylation in YAC128 TG mouse model. These novel results highlight CKD-504 as a potential therapeutic strategy in HD.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.4
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• pp.239-245
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• 2022

Protein and peptide candidates are screened to apply therapeutic application as a drug. Ensuring that these candidates are delivered and maximized effectiveness is still challenging and a variety of studies are ongoing. As drug delivery system vehicles, cell-penetrating peptide (CPP) can deliver various kinds of cargo into the cell cytosol. In a previous study, we developed Ara27 CPP, which are a zinc knuckle family protein of Arabidopsis, and confirmed internalization in human dermal fibroblasts and human dental pulp stem cells at low concentration with short time treatment condition without any toxicity. Ara27, an amphipathic CPP, could be modified and utilized in the biomedical field excluding the risk of toxicity. Therefore, we would like to confirm the non-toxic induced penetrating ability of Ara27 in various cell lines. The purpose of this study was to screen the cell internalization ability of Ara27 in various cell lines and to confirm Ara27 as a promising core CPP structure. First, Ara27 was screened to confirm non-toxicity concentration. Then, fluorescence-labeled Ara27 was treated on human normal cell lines, cancer cell lines and animal cell lines to identify the cellular internalization of Ara27. Ara27 was well intracellular localized in all cell lines and the intensity of fluorescence was remarkably increased in time pass manner. These results indicate that Ara27 has the potential as a core structure for applications in various drug delivery systems.

• Korean Journal of Agricultural Science
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• v.49 no.1
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• pp.137-149
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• 2022

We investigated the effects of Cirsium japonicum var. maackii (CJM) against oxidative stress-induced C6 glial cells and cognitive impairment in mice. To evaluate the anti-oxidative effect of the extract and fractions from CJM, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), reactive oxygen species (ROS), and nitric oxide (NO) assays were conducted in H₂O₂-treated C6 glial cells. Furthermore, we identified the protective mechanisms of CJM with a scopolamine-treated mice model. The results revealed that H₂O₂ decreased the cell viability in C6 glial cells, indicating that H₂O₂ induced oxidative stress in glial cells. However, CJM fractions significantly increased cell viability in H₂O₂-treated C6 glial cells, which suggested that CJM protected against oxidative stress. CJM extract and fractions also reduced ROS and NO production, which were increased by H₂O₂ in C6 glial cells. In particular, the EtOAc fraction from CJM (EACJM) effectively protected against oxidative stress by increasing the cell viability and decreasing ROS and NO. Therefore, we carried out further in vivo experiments with EACJM. Scopolamine caused increases of ROS, thiobarbituric acid reactive substances (TBARS), and NO production. However, EACJM effectively alleviated ROS, TBARS, and NO levels compared to scopolamine-injected mice. In addition, EACJM up-regulated protein expressions of superoxide dismutase and glutathione peroxidase, indicating that EACJM enhanced the antioxidative system. Our results demonstrated that CJM had protective effects against oxidative stress in glial cells and memory dysfunction in mice. Based on these results, we propose that CJM could be a potential AD preventive and therapeutic agent.

• Journal of Convergence Korean Medicine
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• v.4 no.1
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• pp.19-27
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• 2022

Objectives: TSepsis and subsequent acute lung injury (ALI) is a critical state of health caused by infection or endotoxins. This study was conducted to evaluate the effect of Water Extract of Aconiti Lateralis Preparata Radix (AR) on lipopolysaccharide (LPS)-induced sepsis in C57BL/6 mice. Methods: Male C57BL/6 mice were intraperitoneally injected with LPS to induce sepsis and ALI. AR was orally fed twice at 30 min and 180 min after LPS injection. At 24 h post injection, mice were sacrificed, bronchoalveolar lavage fluid (BALF) and blood was collected, and lung tissue was harvested. Hematoxylin and eosin staining was performed in lung tissues, wet/dry ratio of the lung tissue was measured, and the serum cytokine and chemokine levels were analyzed. Results: AR revoked the LPS-induced pathological changes in lung tissues, such as abnormal histological structures, immune cell infiltration and lung edema. Also, AR suppressed the neutrophil infiltration into the lung which was greatly increased by LPS injection based on the cell content of collected BALF. Serum cytokines and chemokines were measured, and AR reversed the LPS-induced increase of cytokines such as interleukin 1 beta, interleukin 6, tumor necrosis factor alpha and chemokines including C-X-C motif chemokine ligand 1 and 2. Conclusion: TAR showed a protective effect in the pathological progress of LPS-induced ALI. Especially, AR suppressed lung edema and infiltration of neutrophils by inhibiting cytokine and chemokine expressions. Such results demonstrate the potential of AR as a therapeutic agent for sepsis and sepsis-induced ALI.

• Journal of Nutrition and Health
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• v.56 no.1
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• pp.12-23
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• 2023

Purpose: This study investigates the alterations in A549 human non-small-cell lung cancer (NSCLC) cells exposed to Citrus junos extract (CJE). We further examine the antiproliferative and apoptotic effects of CJE on NSCLC cells. Methods: Inhibition of proliferation was examined by applying the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) colorimetric assay on CJE-treated A549 NSCLC cells. The lactate dehydrogenase (LDH) assay was performed to measure the degree of toxicity of CJE on NSCLC cells. The effect on migratory proliferation was confirmed using the scratch wound healing assay. The antiproliferative effect of the CJE on human lung cancer cells was verified through morphological observation, fluorescence microscopy, and caspase-3 colorimetry. Results: Exposure of NSCLC cells to CJE resulted in a dose- and time-dependent decrease in cell activity and increased toxicity to the cells. In addition, microscopic observation revealed a reduced ability of the cancer cells to migrate and proliferate after exposure to the CJE, with simultaneous morphological apoptotic changes. Fluorescence staining and microscopic examination revealed that this death was a process of self-programmed cell death of NSCLC cells. Compared to unexposed NSCLC cells, the expression of caspase-3 was significantly increased in cells exposed to CJE. Conclusion: Exposure of A549 human NSCLC cells to CJE inhibits the proliferation, increases the cytotoxicity, and decreases the ability of cells to migrate and grow. Moreover, the expression of caspase-3 increases after CJE treatment, suggesting that the apoptosis of NSCLC cells is induced by a chain reaction initiated by caspase-3. These results indicate that Citrus junos is a potential therapeutic agent for human non-small-cell lung cancer.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.228-236
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• 2023

Background: QiShen YiQi pills (QSYQ) is a Traditional Chinese Medicine (TCM) formula, which has a significant effect on the treatment of patients with myocardial infarction (MI) in clinical practice. However, the molecular mechanism of QSYQ regulation pyroptosis after MI is still not fully known. Hence, this study was designed to reveal the mechanism of the active ingredient in QSYQ. Methods: Integrated approach of network pharmacology and molecular docking, were conducted to screen active components and corresponding common target genes of QSYQ in intervening pyroptosis after MI. Subsequently, STRING and Cytoscape were applied to construct a PPI network, and obtain candidate active compounds. Molecular docking was performed to verify the binding ability of candidate components to pyroptosis proteins and oxygen-glucose deprivation (OGD) induced cardiomyocytes injuries were applied to explore the protective effect and mechanism of the candidate drug. Results: Two drug-likeness compounds were preliminarily selected, and the binding capacity between Ginsenoside Rh2 (Rh2) and key target High Mobility Group Box 1 (HMGB1)was validated in the form of hydrogen bonding. 2 μM Rh2 prevented OGD-induced H9c2 death and reduced IL-18 and IL-1β levels, possibly by decreasing the activation of the NLRP3 inflammasome, inhibiting the expression of p12-caspase1, and attenuating the level of pyroptosis executive protein GSDMD-N. Conclusions: We propose that Rh2 of QSYQ can protect myocardial cells partially by ameliorating pyroptosis, which seems to have a new insight regarding the therapeutic potential for MI.

• BMB Reports
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• v.56 no.8
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• pp.445-450
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• 2023

The development of atherosclerotic cardiovascular disease is associated with the phenotypic switching of vascular smooth muscle cells (SMCs) from a contractile to a synthetic state, leading to cell migration and proliferation. Platelet-derived growth factor-BB (PDGF-BB) modulates this de-differentiation by initiating a number of biological processes. In this study, we show that gene expression of hyaluronic acid (HA) and proteoglycan link protein 1 (HAPLN1) was upregulated during differentiation of human aortic SMCs (HASMCs) into a contractile state, but downregulated upon during PDGF-BB-induced dedifferentiation. This is the first study showing that the treatment of HASMCs with full-length recombinant human HAPLN1 (rhHAPLN1) significantly reversed PDGF-BB-induced decrease in the protein levels of contractile markers (SM22α, α-SMA, calponin, and SM-MHC), and inhibited the proliferation and migration of HASMCs induced by PDGF-BB. Furthermore, our results show that rhHAPLN1 significantly inhibited the phosphorylation of FAK, AKT, STAT3, p38 MAPK and Raf mediated by the binding of PDGF-BB to PDGFRβ. Together, these results indicated that rhHAPLN1 can suppress the PDGF-BB-stimulated phenotypic switching and subsequent de-differentiation of HASMCs, highlighting its potential as a novel therapeutic target for atherosclerosis and other vascular diseases.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.173-179
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• 2009

Objectives : In this study, the effects of different fractions isolated from modified Boyanghwanotang(mBHT) extract on LPS-induced inflammation in BV2 microglial cells were investigated. Methods : mBHT was extracted with water, and then fractionated with n-hexane, methylene chloride, ethylacetate and n-butanol. BV2 cells, a mouse microglia line were incubated with different concentrations of each fraction of mBHT for 30 min, and then stimulated with LPS for 24 h. Cell toxicity was determined by MTT assay. The concentration of nitric oxide (NO) was measured in culture medium by Griess reagent assay. The expression of inducible nitric oxide synthease (iNOS) protein was determined by Western blot. Results : Four fractions of mBHT were significantly inhibited LPS-induced NO productions in BV2 cells in a dose-dependent manner. The methylene chloride fraction of mBHT was most strongly inhibited the NO production compared with those of the others. The methylene chloride fraction of mBHT was also suppressed LPS-induced iNOS expression comparison of other fractions at same concentration ($50\;{\mu}g/ml$) in BV2 cells. Conclusions : The results showed that the methylene chloride fraction of mBHT may have an strong anti-inflammatory property through the inhibition of NO production and iNOS expression in activated microglia, and could a therapeutic potential for the treatment of various brain inflammatory diseases.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.169-178
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• 2009

Objectives : In this study, the effects of ethylacetate extract of Ostericum koreanum on inflammation in RAW264.7 cells were investigated. Methods : Dried roots of Ostericum koreanum was extracted with 80% methanol for 24 h, and then fractionated with n-butanol, n-hexan and ethylacetate. RAW264.7 cells, a mouse macrophage line were incubated with different concentrations of the extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin E2 ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expressions of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX) -2 mRNA and protein were determined by RT-PCR and Western blot. Results : The methanol extract of Ostericum koreanuman and its fractions were significantly inhibited the NO and PGE2 productions in LPS-stimulated RAW264.7 cells. Among the fractions of Ostericum koreanuman the ethylacetate fraction was more strongly inhibited NO and $PGE_2$ productions compared with other fractions. The ethylacetate fraction was also suppressed LPS-induced mRNA expressions of iNOS and its protein levels in RAW264.7 cells. Conclusions : This study suggests that the ethylacetate fraction of Ostericum koreanum may have an anti-inflammatory property through suppressing inflammatory mediator productions in activated macrophages, suggesting have a therapeutic potential for the treatment of various inflammatory diseases.

• Biomedical Science Letters
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• v.29 no.1
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• pp.11-25
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• 2023

In hepatocellular carcinoma (HCC), chromosome 4 open-reading frame 47 (C4orf47) has not been so far investigated for its prognostic value or association with infiltrating immune cells. We performed bioinformatics analysis on HCC data and analyzed the data using online databases such as TIMER, UALCAN, Kaplan-Meier plotter, LinkedOmics, and GEPIA2. We found that C4orf47 expression in HCC was higher compared to normal tissues. High C4orf47 expression was associated with a worse prognosis in HCC. The correlation between C4orf47 and infiltrating immune cells is positively associated with CD4+T cells, B cells, neutrophils, macrophages, and dendritic cells in HCC. Moreover, high C4orf47 expression was correlated with a poor prognosis of infiltrating immune cells. Analysis of C4orf47 gene co-expression networks revealed that 12501 genes were positively correlated with C4orf47, whereas 7200 genes were negatively correlated. The positively related genes of C4orf47 are associated with a high hazard ratio in different types of cancer, including HCC. Regarding the biological functions of C4orf47 gene, it mainly regulates RNA metabolic process, DNA replication, and cell cycle. The C4orf47 gene may play a prognostic role by regulating the global transcriptome process in HCC. Our findings demonstrate that high C4orf47 expression correlates with poor prognosis and tumor-infiltrating immune cells in HCC. We suggest that C4orf47 is a novel prognostic biomarker and potential immune therapeutic target for HCC.