• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.665-683
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• 2002

A novel glucanhydrolase from a mutant of Lipomyces starkeyi KSM 22 has additional amylase activity besides mutanolytic activity and has been suggested as promising anti-plaque agent. It has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 glucanhydrolase are desirable for its application as a dental plaque control agent. In human experimental gingivitis　model and 6 month clinical trial, mouthrinsing with Lipomyces　starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effect was negligible. This study was aimed to evaluate the cytotoxic effect of Lipomyces starkeyi KSM 22 glucanhydrolase on human gingival fibroblasts. Primary culture of human gingival fibroblasts at the 4th to 6th passages were used. Glucanhydrolase solution was made from lyophilized glucanhydrolase powder from a mutant of Lipomyces stakeyi KSM 22 solved in PBS and added to DMEM medium to the final concentration of 0.5, 1, and 2 unit. Cells were exposed to glucanhydrolase solution or 0.1 % chlorhexidine and the cells cultured in DMEM with 10% FBS and 1% antibiotics as control. After exposure, the morphological change, cell attachment, and cell activity by MTT assay were evaluated in 0.5, 1.5, 3, 6, 24 hours after treatment. The cell proliferation and cell activity was also evaluated at 2 and 7 days after 1 minute exposure, twice a day. The cell morphology was similar between the Lipomyces smkeyi KSM 22 glucanhydrolase groups and control group during the incubation periods, while most fibroblasts remained as round cell regardless of incubation time in the chlorhexidine group. The numbers of the attached cells in the glucanhydrolase groups were comparable to that of control and significantly higher than the chlorhexidine group. The numbers of the proliferated cells in the glucanhydrolase groups at 7 days of incubation were comparable to the control group and higher than the chlorhexidine group. The cell activity in glucanhydrolase groups paralleled with the increased cell number by attachment and proliferation. According to these results, Lipomyces starkeyj KSM 22 glucanhydrolase has little harmful effect on attachment and proliferation of human gingival fibroblasts, in contrast to 0.1% chlorhexidine which was cytotoxic to human gingival fibroblasts. Therefore this glucanhydrolase preparation is considered as a safe and promising agent for new mouthwash formula in the near future.

• Korean Journal of Food Science and Technology
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• v.40 no.6
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• pp.696-701
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• 2008

The objective of this study was to determine whether Codonopsis lanceolata or Platycodon grandiflorum ethyl acetate fraction (CLEA or PGEA) protect cells against sodium nitroprusside (SNP)-induced oxidative stress via the expression of various antioxidant systems. The HepG2 cells exposed for 24 hr to 0.5 mM SNP showed a reduction in the cell viability by an MTT assay. Pretreatment with CLEA and PGEA resulted in an inhibition of SNP-induced cell death. In addition, the effects of CLEA and PGEA on the expression of antioxidant systems via RT-PCR analyses was assessed. The levels of catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD) and metallothionein (MT)-1A mRNA were increased after 24 hr of CLEA exposure. The levels of Mn superoxide dismutase CAT, G6PD, MT-1A, and MT-2A mRNA were increased after PGEA treatment. In conclusion, CLEA and PGEA exert indirect antioxidant effects, perhaps via the induction of a variety of antioxidant systems which, may protect cells against oxidative stress.

• Journal of Life Science
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• v.18 no.12
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• pp.1644-1650
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• 2008

The essential oil of tea tree (Melaleuca alternifolia) is widely used in traditional Australian medicine for skin lesions and infected injuries. In the present study, we investigated the chemical composition, cytotoxicity and its biological activities. The composition of the oil was analyzed by GC-MS. ${\beta}$-Terpinene (20.87%), ${\alpha}$-pinene (17.60%), p-cymene (11.23%), 3-carene (10.40%), trans-anethole (8.47%) and limonene (4.65%) were the major components in the oil. The results tested by MTT assay indicated that the oil showed no cytotoxic effect, at concentrations up to 5%, for less than 3h. The antiradical capacity was evaluated by measuring the scavenging activity of the essential oil on the 2,20-diphenylpicrylhydrazyl (DPPH) and 2,2'-azino-bis 3-ethyl benzothiazoline-6-sulfonic acid (ABTS) radicals. The oil was able to reduce the both radicals dose-dependently, and the concentration required for 50% reduction ($RC_{50}$) against ABTS radicals ($1.6{\pm}0.02%$) was slightly lower than DPPH radicals ($2.6{\pm}0.29%$). The direct contact and vapor-phase antibacterial activity of the oil were also evaluated using disc diffusion method against Staphylococcus aureus, Streptococcus mutans, Listeria monocytogenes, Acinetobacter baumannii, Escherichia coli, and Vibrio parahaemolyticus. All the Gram-negative bacterial strains tested showed more sensibility to the oil than the Gram-positive strains when compare to the effect of gentamycin. On the other hand, the vapor phase of the essential oil against S. aureus exhibited strongest inhibitory effect.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.8 no.1
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• pp.103-125
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• 2002

This experimental study was carried out to evaluate the anti-tumor and the immunomodulatory effects of Jiaweicitaowan(加未慈桃丸) against cancer. The in vitro anti-tumor effects were evaluated by MTT assay. The cytotoxicity, extension of survival days, the effect of inhibition solid tumor which was induced sarcoma 180, and the changes of body weight were evaluated for in vivo effects of anti-tumor. To evaluate the immunomodulatory effects of Jiawei- citaowan(加未慈桃丸), delayed type hypersensitivity, hemagglutinin, hemolysin titers for humoral immune response, rosette forming cells for cell-mediated immune response, natural killer cell activity, proliferation of lymphocyte, productivty of Interleukin-2, and carbon clearance were measured with methotrexate treated mice. The results were as follows; 1. In the case of existence ability of tumor cell, IC50 had an anti-tumor ativity resulted 2.52mg/ml to SNU-C4. 0.41mg/ml to SNU-396, resulted to 0.09mg/mlSNU-1. 2. The groups of Jiaweicitaowan(加未慈桃丸) 10mg/ml, 20mg/kg had no body weight loss. reduction in intake of water and feed, so these had no toxicity. 3. In the case of the effect of extention of existence. the group of 20mg/kg Jiaweicitaowan(加未慈桃丸) extract treated group was showed 250% in ILS. 4. The effect of inhibition solid tumor was significantly decreased in both 10mg/kg, 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group S. The groups of 10mg/kg, 20mg/kg Jiaweicitaowan(加未慈桃丸) had significant effect of body weight change compared to control group. 6. Delayed type hypersensitivity was not significant in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 7. Hemagglutinin and Hemolysin titers were significantly increased by dose-dependent. so these results showed that the humoral immume respose was activated. 8. For the effect of rosette formimg cells was not significant in hoth Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 9. Natural killer cell activity was significantly increased in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group in the ratio of 100: 1, 50: 1 of effector and target cells, but in the ratio of 10:1, the Jiaweicitaowan(加未慈桃丸) extract treated groups were not significant. 10. The proliferation of lymphocyte and productivty of Interleukin-2 were significantly increased by dose-dependent in both 10mg/ kg, 20mg/ kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 11. In the phagocytic effect, the 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated group showed the increasing effect with significance as compared with control group. According to the results, we can suggest that Jiaweicitaowan(加未慈桃丸) has the antitumor and the immunomodulatory effects.

• Journal of the East Asian Society of Dietary Life
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• v.24 no.6
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• pp.776-784
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• 2014

Korean dishes, Hansik are characterized by healthful vegetable intake. The purpose of this study was to evaluate the inhibitory effect of commonly consumed vegetables by Koreans on obesity/metabolic disease-related inflammation. Through statistical analysis of the KNHANES database ($1^{st}$ 1998, $5^{th}$ 2010, 2011) and a literature review, we selected vegetables for study. Among the vegetables, main or sub ingredients of Kimchi were excluded. Samples were prepared using only edible portions and freeze-dried. After grinding, samples were extracted with ethanol, evaporated and finally lyophilized. The cytotoxicity of samples was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, at various concentrations that do not affect cell viability. Raw 264.7 macrophages were treated with lipopolysaccharide (LPS) and 11 kinds of samples or positive control (troglitazone) dissolved in dimethyl sulfoxide (DMSO). After 24 hours, nitric oxide (NO), tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) and monocyte chemoattractant protein-1 (MCP-1) production were determined. Excepts for young pumpkin and bracken, nine samples effectively reduced NO production compared with control treated with LPS and DMSO. NO levels of five samples (bean sprouts, leeks, eggplant, mugwort, and pumpkin) were similar to that of the positive control. These five samples showed significantly decreased TNF-${\alpha}$ or MCP-1 compared to the control group. Our results suggest that consumption of commonly consumed vegetables contributes to partial prevention of obesity and related metabolic syndrome through reduction of NO, TNF-${\alpha}$, and MCP-1 production.

• The Journal of Korean Obstetrics and Gynecology
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• v.23 no.3
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• pp.38-55
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• 2010

Purpose: This study was designed to find out the anti-cancer effects of Samultang-Gami which was composed of Rehmanniae Radix(RR), Angelicae Gigantis Radix(AGR), Cnidii Rhizoma(CR), Paeoniae Radix(PR), Cortex Moutan Radicis(CMR), Hedyotis Diffusa(HD) and Caesalpinia Sappan on HeLa, HepG2 and AGS cells. Methods: Various cancer cell lines including HeLa, HepG2 and AGS cells, were used. In vitro anti-cancer effects were measured by MTT assay using cancer cell lines treated with various concentrations of 70% ethanol extract of Samultang-Gami. Expression of cell cycle arrest mediators including Bax, Bcl-2, p53 and DARP-1 proteins were measured by Western blot analysis. Results: 1. Samultang-Gami decreased the viability of HeLa and HepG cells in a dosedependent manner. 2. AGR, CMR, PR and HD decreased the viability of HeLa, HepG2 and AGS cells. 3. We could observe that the decreased Bax and Bcl-2 expression level and the increased PARP-1 expression level by Samultang-Gami extracts treated in HeLa cells. 4. We could observe that the decreased Bcl-2 expression level and the increased Bax, p53 and PARP-1 expression level by RR extracts treated in HeLa cells. and also could observe that the reduction of the protein level of Bcl-2, p53 and PARP-1 and the increase of the protein level of Bax by PR in HeLa cells. 5. We could observe that the increased p53 expression level, the decreased PARP-1's that and the unchanged Bax and Bcl-2's that by Samultang-Gami extracts treated in HepG2 cells. 6. We could observe that the reduced Bcl-2 expression level by each of RR extracts and PR extracts in HepG2 cells. 7. The treatment of Samultang-Gami in AGS cells didn't have any effect on the expression level of Bax, Bcl-2, p53 and PARP-1. 8. We could observe that the increased p53 and PARP-1 expression level by each of CR, RR and PR extracts in AGS cells. Conclusion: Taken together, we suggest that Samultang-Gami exhibits cytotoxic effects on HeLa, HepG2 and AGS cells, causing apoptosis. The results showed that Samultang-Gami may do so by regulating the expression of specific target molecules that promote efficient apoptotic cell death in a dose-dependent manner.

• Korean Journal of Acupuncture
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• v.24 no.1
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• pp.171-187
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• 2007

Objectives : This study was performed to determine if Orostachys japonicus A. Berger herbal acupuncture (OjB) provides the protective effect against the loss of cell viability and DNA damage induced by oxidant in renal proximal tubular cells. Methods : The cell viability was evaluated by a MTT reduction assay and DNA damage was estimated by measuring double stranded DNA breaks in opossum kidney (OK) cells, an established proximal tubular cell line. Lipid peroxidation was determined by measuring malondialdehyde (MDA), a product of lipid peroxidation. Results : H2O2 increased the loss of cell viability in a time-dependent manner, which were prevented by 0.1% OjB. The protective effect of OjB was dose-dependent over concentration range of 0.05-0.5%. H2O2 caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by H2O2 was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Generation of superoxide and H2O2 in neutrophils activated by phorbol-12,13-dibutyrate was inhibited by OjB in a dose-dependent manner. OjB inhibited generation of H2O2 in OK cells treated with antimycin A and exerted a direct H2O2 scavenging effect. Exposure of OK cells to 1 mM tBHP caused a significant depletion of glutathione which was prevented by OjB. OjB accelerated the recovery in cells cultured for 20 hr in normal medium without oxidant following oxidative stress. Conclusions : These results suggest that OjB exerts the protective effect against oxidant-induced cell injury and its protective effect was resulted from radical scavenging and antioxidant activities.

• International Journal of Oral Biology
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• v.39 no.2
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• pp.97-105
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• 2014

The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

• Food Science and Preservation
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• v.22 no.5
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• pp.751-757
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• 2015

The objective of this study was to investigate the protective effects of mulberry (Morus alba) sugar extracts (MSE) against $H_2O_2$-induced oxidative stress in HepG2 cells. The MSEs was mixed with matured mulberry and sugar at the same ratio (1:1, w/w) and stored at $18{\pm}3^{\circ}C$ for 40 days. In 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging test, MSE stored for 40 days showed high activity with a ratio above 66%. Therefore, we selected 40 days as the optimum storage period. After cell viability analysis using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, we determined that the optimum concentration of MSE was 0.5%. Our results showed that MSE increased the cell viability and antioxidant enzyme activities of superoxide dismutase (SOD) and catalase in $H_2O_2$-treated HepG2 cells. Moreover, the treatment with MSE inhibited malondialdehyde (MDA) levels in $H_2O_2$-treated HepG2 cells. We also observed a reduction in apoptotic bodies in the Hoechst staining. These data show that MSE treatment significantly suppressed caspase-3 activity in HepG2 cells expored to $H_2O_2$-induced oxidative stress, thereby indicationg the protective effects of MSE in $H_2O_2$-induced oxidative stress.

• Journal of Life Science
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• v.16 no.4
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• pp.659-663
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• 2006

The extracts of Plocamium telfairiae using several solvents with different polarities were prepared and their growth inhibitory effects were examined on the human cancer cells. We investigated the cytotoxic effects of P. telfairiae extracts on HT-29 cells by the MTT reduction assay and examining the morphological change under the inverted microscope. Among three extracts, the methanol extract showed the strongest inhibitory effect on the growth of HT-29 cells. The methanol extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and aqueous for purifying crude methanol extract. The n-hexane layer among the fractioned layers showed remarkable inhibitory activity on the growth of HT-29 cells. Moreover n-hexane layer showed the notable growth inhibition effects with a dose-dependent manner against SW620, HeLa, and MCF-7 cells as well as HT-29 cells. These results indicated that P. telfairiae extracts may be contained bioactive materials with inhibitory effect on the growth of human cancer cells.