• Biomolecules & Therapeutics
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• v.23 no.5
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• pp.414-420
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• 2015

Flavonoids, such as fisetin (3,7,3',4'-tetrahydroxyflavone), are plant secondary metabolites. It has been reported that fisetin is able to perform numerous pharmacological roles including anti-inflammatory, anti-microbial, and anti-cancer activities; however, the exact anti-inflammatory mechanism of fisetin is not understood. In this study, the pharmacological action modes of fisetin in lipopolysaccharide (LPS)-stimulated macrophage-like cells were elucidated by using immunoblotting analysis, kinase assays, and an overexpression strategy. Fisetin diminished the release of nitric oxide (NO) and reduced the mRNA levels of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-${\alpha}$, and cyclooxygenase (COX)-2 in LPS-stimulated RAW264.7 cells without displaying cytotoxicity. This compound also blocked the nuclear translocation of p65/nuclear factor (NF)-${\kappa}B$. In agreement, the upstream phosphorylation events for NF-${\kappa}B$ activation, composed of Src, Syk, and I${\kappa}B{\alpha}$, were also reduced by fisetin. The phospho-Src level, triggered by overexpression of wild-type Src, was also inhibited by fisetin. Therefore, these results strongly suggest that fisetin can be considered a bioactive immunomodulatory compound with anti-inflammatory properties through suppression of Src and Syk activities.

• Biomolecules & Therapeutics
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• v.21 no.3
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• pp.229-233
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• 2013

This study was aimed at investigating the possible effects of phytoceramide (Pcer) on learning and memory and their underlying mechanisms. Phytoceramide was orally administered to ICR mice for 7 days. Memory performances were assessed using the passive avoidance test and Y-maze task. The expressions of phosphorylated cAMP response element binding protein (pCREB), brain-derived neurotrophic factor (BDNF) were measured with immunoblot. The incorporation of 5-bromo-2-deoxyuridine (BrdU) in hippocampal regions was investigated by using immunohistochemical methods. Treatment of Pcer enhanced cognitive performances in the passive avoidance test and Y-maze task. Immunoblotting studies revealed that the phosphorylated CREB and BDNF were significantly increased on hippocampus in the Pcer-treated mice. Immunohistochemical studies showed that the number of immunopositive cells to BrdU was significantly increased in the hippocampal dentate gyrus regions after Pcer-treatment for 7 days. These results suggest that Pcer contribute to enhancing memory and BDNF expression and it could be secondary to the elevation of neurogenesis.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.02a
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• pp.55-56
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• 2001

Gelatin zymograms of bFF and bS showed GA110 and 62 kDa gelatinses in adsition to several minor ones. Of these, GA110 gelatinase was abolished by treating bFF or bS with bOF and interestingly, its enzymatic activity was enhanced by adding EDTA to bFF or bS before zymographic analyses. Experiments using specific inhibitors of MMPs indicated that GA110 and 62 kDa proteins were indeed gelatinases. Immunoblotting experiments using an antibody against human MMP-2 showed that both GA110 and 62 kDa were an MMP-2 isoform and active MMP-2, respectively. The results suggest that the interaction between bFF and bOF can occur at the time of fertilization.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.59-68
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• 2017

Caffeoyl shikimate esterase (CSE) is a key enzyme in lignin synthesis in Arabidopsis thaliana. To determine the role of CSE in lignification of the endocarp in peach (Prunus persica L.) fruit, we cloned and characterized the P. persica CSE homolog, which we designated PpCSE. The 954 - bp PpCSE gene encoded a 317 - amino acid polypeptide. PpCSE expression patterns in the mesocarp and endocarp changed during peach fruit development. There was no significant difference between the expression levels of PpCSE in the mesocarp and endocarp at 39 and 44 days after full bloom (DAFB), but the expression level of PpCSE in the endocarp at 50 and 55 DAFB was 80.73 and 72.75 times higher, respectively, than that in the mesocarp. During peach fruit development, PpCSE expression in the endocarp increased rapidly; the relative PpCSE expression level at 50 DAFB was 122.70 times higher than that at 39 DAFB. At the protein level, CSE was detected in the peach fruit endocarp at 50 and 55 DAFB. Our study suggests that PpCSE expression during peach fruit development is closely related to the degree of endocarp lignification.

• Development and Reproduction
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• v.24 no.2
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• pp.101-111
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• 2020

Coiled-coil domain containing 110 (CCDC110, KM-HN-1) is a protein containing C-terminal coiled-coil domain (CCD) which was previously discovered as a member of the human cancer/testis antigen (CTA). In addition, CCDC110 has both nuclear localization signal sequence and the leucine zipper motif. Although the functional role of CCDC110 has yet to be fully identified, the mRNA expression levels of CCDC110 are known to be highly elevated in various cancer types including testis, implying its relevance to cancer pathogenesis. In this study, we first developed several monoclonal antibody (mAb) hybridoma clones targeting CCDC110 and further isolated clone by characterizing for its specificity using immunoblotting and immunoprecipitation approaches with basal parenchymal sperm cells in testis tissue. Next, using these mAbs, we showed that the Tet-inducible overexpression of CCDC110 protein delayed the entry of G2/M phase in U2-OS osteosarcoma cells. Based on these results, we propose that CCDC110 plays a crucial role in cell cycle progression.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.259-267
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• 2005

The present study was conducted to investigate gelatinolytic activities in HAM and to determine whether there are any changes in gelatinolytic activity profiles when the cells are cultured in hepatogenic medium. Placenta was obtained during caesarean section of the volunteers, with informed consent. HAM were isolated from amniotic membrane using collagenase type A HAM were cultured in hepatogenic medium for 3 weeks and the conditioned media were obtained at day 7, 14 and 21. The zymographic pattern of gelatinolytic activity of the HAM did not undergo a change during passages. When the HAM were cultured in a fibronectin-coated dishes in a hepatogenic medium, there was no significant difference of the gelatinase pattern between before and after culture. However, when bFGF was added to the culture, a dramatic increase of 62kDa and 59kDa gelatinases was observed. Interestingly, when ITS instead of FN was present, HAM-conditioned medium also showed a similar increase of both gelatinases. Immunoblotting analysis demonstrated that both 62kDa and 59kDa gelatinases were the active form of MMP-2 resulting from the turnover of MMP-2 proform. Futher study will be necessary to determine the relationship between bFGF and active MMP-2 during hepatogenesis of HAM.

• Journal of Periodontal and Implant Science
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• v.41 no.3
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• pp.157-163
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• 2011

Purpose: Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. Methods: LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. $I{\kappa}B-{\alpha}$ degradation, nuclear translocation of NF-${\kappa}B$ subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-${\kappa}B$ was also analyzed. Results: Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of $I{\kappa}B-{\alpha}$ induced by P. intermedia LPS. Curcumin blocked NF-${\kappa}B$ signaling through the inhibition of nuclear translocation of NF-${\kappa}B$ p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. Conclusions: Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

• Korean Journal of Plant Resources
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• v.28 no.3
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• pp.321-328
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• 2015

Alpiniae officinarum Rhizoma (the rhizome of Alpinia officinarum Hance, known as lesser galangal), a family of Zingiberaceae, has been used to reduce pain of infection and inflammatory diseases in Asian countries. The present study was focused to evaluate the inhibitory degranulation effect of Alpiniae officinarum Rhizoma extract in RBL-2H3 rat basophilic leukemia cells. Cell viability was measured by MTT assay. RBL-2H3 cells were stimulated by phorbol 12-myristate 13-acetate and calcium ionophore A23187. Mast cell degranulation was analyzed by measuring release of β-hexosaminidase in RBL-2H3 cell. Gene expression was measured by qRT-PCR and signaling molecules were detected by immunoblotting. The Alpiniae officinarum Rhizoma extract suppressed β-hexosaminidase release in dose-dependent manner and inhibited cycloxygenase-2 and tumor necrosis factor-α gene expression. Furthermore, it was found that Alpiniae officinarum Rhizoma extract reduced mitogen-activated protein kinases, especially phosphorylated p38, at 0.75 ㎎/㎖ of Alpiniae officinarum Rhizoma extract concentrations. These data show that Alpiniae officinarum Rhizoma extract has immunosuppressive effect in mast cell induced allergic inflammation.

• YAKHAK HOEJI
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• v.45 no.1
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• pp.39-45
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• 2001

The pork meat has been reported as one of the food occurring allergic reactions predominantly to korean. To identify the potential food allergens in pork meat, sera were collected from 25 allergic patients to the pork meat and 10 allergic patients not to pork meat as well as 5 normal subjects after skin prick test and open food challenge test. Crude extracts were prepared by blending raw pork meat in phosphate buffered saline (pH 7.0) and the heat treatment on crude extracts was carried to characterize sensibility of the allergens to heat. ELISA was performed to determine specific IgE antibody levels of allergic patients to pork meat, and resulted in twofold higher mean value than that of tolerated patients. Extracted proteins from pork meat was separated with SDS-PAGE followed by immunoblotting using sera from pork sensitive patients and control subjects, respectively. The IgE binding response to pork meat by immunobots correlated with quantitative specific IgE value of each person. Immunoblots showed four prominent IgE-binding bands (66, 60, 50, 44 kDa) in crude extract, but two bands of those (60, 44 kDa) were heat-labile. These results suggest that most prominent allergens from pork meat are four components(66, 60, 50, 44 kDa) in korean and the heat treatment on allergen is additional parameter to characterize allergen.

• Journal of Ginseng Research
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• v.38 no.4
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• pp.251-255
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• 2014

Background: Ginsenoside Rp1 (G-Rp1) is a novel ginsenoside derived from ginsenoside Rk1. This compound was reported to have anticancer, anti-platelet, and anti-inflammatory activities. In this study, we examined the molecular target of the antiproliferative and proapoptotic activities of G-Rp1. Methods: To examine the effects of G-Rp1, cell proliferation assays, propidium iodine staining, proteomic analysis by two-dimensional gel electrophoresis, immunoblotting analysis, and a knockdown strategy were used. Results: G-Rp1 dose-dependently suppressed the proliferation of colorectal cancer LoVo cells and increased their apoptosis. G-Rp1 markedly upregulated the protein level of apolipoprotein (Apo)-A1 in LoVo, SNU-407, DLD-1, SNU-638, AGS, KPL-4, and SK-BR-3 cells. The knockdown of Apo-A1 by its small-interfering RNA increased the levels of cleaved poly(ADP-ribose) polymerase and p53 and diminished the proliferation of LoVo cells. Conclusion: These results suggest that G-Rp1 may act as an anticancer agent by strongly inhibiting cell proliferation and enhancing apoptosis through upregulation of Apo-A1.