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Kim C.-D.,Sychar Maxface Center for Maxillofacial Surgery | Cha J.-D.,Institute of Jinan Red Ginseng | Li S.,Yanbian University | Cha I.-H.,Yonsei University
Archives of Oral Biology

Background Acacetin (5,7-dihydroxy-40-methoxyflavone), present in safflower seeds, plants, flowers, Cirisium rhinoceros Nakai, has been reported to be able to exert anti-peroxidative, anti-inflammatory, anti-plasmodial, and anti-proliferative activities by inducing apoptosis and blocking the progression of cell cycles. Objective and design The objective of this study is to investigate the mechanism of acacetin-induced apoptosis of oral squamous cell carcinoma cell line (HSC-3). Results Acacetin caused 50% growth inhibition (IC50) of HSC-3 cells at 25 μg/mL over 24 h in the MTT assay. Apoptosis was characterized by DNA fragmentation and increase of sub-G1 cells and involved activation of caspase-3 and PARP (poly-ADP-ribose polymerase). Maximum caspase-3 activity was observed with 100 μg/mL of acacetin for 24 h. Caspase-8 and -9 activation cascades mediated the activation of caspase-3. Acacetin caused reduction of Bcl-2 expression leading to an increase of the Bax:Bcl-2 ratio. It also caused a loss of mitochondrial membrane potential that induced release of cytochrome c into the cytoplasm. Pretreatment with casapse-3 (Z-DEVD-FMK), -8 (Z-IETD-FMK), and 9 inhibitor (z-LEHD-fmk) inhibited the acacetin-induced loss of mitochondrial membrane potential and release of cytochrome c. The mitogen-activated protein kinases (MAPKs) were activated by acacetin. Moreover, pretreating the cells with each of the caspase inhibitor or MAPKs specific inhibitors apparently inhibited acacetin-induced cytotoxicity of HSC-3 cells. Conclusion In conclusion, acacetin induce the apoptosis of oral squamous cell carcinoma cell line, which is closely related to its ability to activate the MAPK-mediated signaling pathways with the subsequent induction of a mitochondria- and caspase-dependent mechanism. These results strongly suggest that acacetin might have cancer inhibition and therapeutic potential. © 2015 Elsevier Ltd Elsevier Ltd. All rights reserved. Source

Cha S.-M.,Chonbuk National University | Cha J.-D.,Institute of Jinan Red Ginseng | Jang E.-J.,Daegu Health College | Kim G.-U.,Pohang College | Lee K.-Y.,Chonbuk National University
Archives of Oral Biology

Background Sophora flavescens AITON (Leguminosae) is a typical traditional Korean medical herb considered to exhibit antibacterial, anti-inflammatory, and antipyretic effects, and is also used for the treatment of skin and mucosal ulcers, sores, diarrhea, gastrointestinal hemorrhage, arrhythmia, and eczema. Objective and design This study examined the inhibitory effects of sophoraflavanone G (SF) of S. flavescens on the bacterial fibrillar protein, Antigen I/II (AgI/II)-N recombinant protein isolated from Streptococcus mutans(rAg I/II)-induced production of nitric oxide (NO) and prostaglandin E2 (PGE2). The investigation was focused on whether SF could inhibit the production of proinflammatory mediators such as nitric oxide (NO) and prostaglandin (PG) E2 as well as the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-a, interleukin (IL)-6, nuclear factor (NF)-κB and mitogen-activated protein kinases (MAPKs) in rAgI/II-stimulated RAW 264.7 cells using Griess reagent, Enzyme linked immunosorbent assay (ELISA), and Western blotting analysis. Results SG significantly inhibited the production of NO and PGE2 and pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor α in Ag I/II-N-stimulated RAW264.7 cells, which were mediated by the down-regulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. The SF inhibited the phosphorylation of IκB-α, nuclear translocation of p65, and subsequent activation of NF- κB in the rAgI/II-stimulated cells. In addition, the SF suppressed the rAgI/II-stimulated activation of ERK MAPK as well as the MAPK inhibitor significantly reduced the rAgI/II-induced production of NO and PGE2. Conclusion Collectively, we suggest that the SF inhibits the expression and production of inflammatory mediators by blocking the ERK MAPK mediated pathway and inhibiting the activation of NF-κB. © 2016 Elsevier Ltd. All rights reserved. Source

Sung Y.-Y.,Korea Institute of Oriental Medicine | Kim S.-H.,Daejeon University | Yoo B.W.,Institute of Jinan Red Ginseng | Kim H.K.,Korea Institute of Oriental Medicine
BMC Complementary and Alternative Medicine

Background: In traditional oriental medicine, A. fistulosum and V. mandshurica are considered to be effective in promoting blood circulation. Therefore, in this study, we investigated whether a solution containing both A. fistulosum and V. mandshurica (AFE + VME) extracts has synergistic effects on the treatment of hyperlipidemia and obesity. Methods: Anti-obesity effects of an herbal extract containing Allium fistulosum and Viola mandshurica (AFE + VME) were investigated in high-fat diet (HFD)-induced obese mice. AFE + VME was orally administrated to mice with the HFD at a dose of 200mg/kg/day for 8weeks. We observed the effects of mixed extract on body weight, fat mass, serum lipid levels, and mRNA expression levels of lipid metabolism-related genes in the adipose tissue of mice. Results: The nutritional analysis revealed that this mixed extract is high in carbohydrate (72.2g/100g) and protein (11.5g/100g); low in fat (1.7g/100g); rich in vitamins E (4.8mg/100g), B1 (14.8mg/100g), B2 (1.0mg/100g), niacin (7.9mg/100g), and folic acid (1.57mg/100g); and rich in minerals such as calcium (600mg/100g), iron (106.1mg/100g), and zinc (5.8mg/100g). The oral administration of AFE + VME in obese mice reduced body weight, tissue weight, adipocyte size, and lipid accumulation in the liver compared with HFD control mice. AFE + VME also decreased serum triglyceride, total cholesterol, and leptin concentrations. Furthermore, AFE + VME markedly increased the mRNA expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), uncoupling protein-2 (UCP-2), and adiponectin and decreased leptin expression in the epididymal white adipose tissue. Our results suggest that the extract containing A. fistulosum and V. mandshurica improved lipid metabolism via the up-regulation of PPAR-γ, UCP-2, and adiponectin expression and the down-regulation of leptin in HFD-induced obese mice. Conclusions: Therefore, the extract containing Allium fistulosum and Viola mandshurica may be a potentially effective therapy for obesity and its related metabolic disorders such as hyperlipidemia and insulin resistance. © 2015 Sung et al. Source

Ki M.R.,Daegu University | Yun S.K.,Korea University | Choi K.M.,Institute of Jinan Red Ginseng | Hwang S.Y.,Korea University
Journal of Microbiology and Biotechnology

An acid phosphatase (HppA) activated by NH 4Cl was purified 192- and 34-fold from the periplasmic and membrane fractions of Helicobacter pylori, respectively. SDS-polyacrylamide gel electrophoresis revealed that HppA from the latter appears to be several kilodaltons larger in molecular mass than from the former by about 24 kDa. Under acidic conditions (pH≤4.5), the enzyme activity was entirely dependent on the presence of certain mono- and/or divalent metal cations (e.g., K +, NH 4 +, and/ or Ni 2+). In particular, Ni 2+ appeared to lower the enzyme's Km for the substrates, without changing Vmax. The purified enzyme showed differential specificity against nucleotide substrates with pH; for example, the enzyme hydrolyzed adenosine nucleotides more rapidly at pH 5.5 than at pH 6.0, and vice versa for CTP or TTP. Analyses of the enzyme's N-terminal sequence and of an HppA - H. pylori mutant revealed that the purified enzyme is identical to rHppA, a cloned H. pylori class C acid phosphatase, and shown to be the sole bacterial 5'-nucleotidase uniquely activated by NH 4Cl. In contrast to wild type, HppA - H. pylori cells grew more slowly. Strikingly, they imported Mg 2+ at a markedly lowered rate, but assimilated urea rapidly, with a subsequent increase in extracellular pH. Moreover, mutant cells were much more sensitive to extracellular potassium ions, as well as to metronidazole, omeprazole, or thiophenol, with considerably lowered MIC values, than wild-type cells. From these data, we suggest that the role of the acid phosphatase HppA in H. pylori may extend beyond 5'-nucleotidase function to include cation-flux as well as pH regulation on the cell envelope. © The Korean Society for Microbiology and Biotechnology. Source

Sung Y.-Y.,Korea Institute of Oriental Medicine | Kim S.-H.,Daejeon University | Kim D.-S.,Korea Institute of Oriental Medicine | Park S.H.,Korea Institute of Oriental Medicine | And 2 more authors.
Journal of Functional Foods

The nutritional composition and anti-obesity activities of cereal bars containing Allium fistulosum extract (AFB) in high-fat diet-induced obese mice was determined. AFB is rich in vitamins C, B2, and B3, and B9 with high protein. Oral treatment of AFB in obese mice reduced body weight, lipid accumulation in liver and adipose tissue as well as adipocyte size, compared to the high fat diet control mice. AFB also decreased serum triacylglycerol, glucose, and insulin concentrations, with increased HDL-cholesterol and adiponectin levels. Furthermore, 5% AFB markedly increased mRNA expression of peroxisome proliferators activated receptor-γ, uncoupling protein-2 as well as β3-adrenoreceptor in the visceral adipose tissue. These results demonstrate that the cereal bar with A. fistulosum extract may potentially serve as nutraceutical for improvement of obesity and metabolic disorders. © 2013 Elsevier Ltd. Source

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