• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.6
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• pp.636-642
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• 2007

Background: CpG DNA plays an important role in immune cell function. This study examined whether the temporal control of toll-like receptor (TLR)9 by CpG DNA can regulate the expression of matrix metalloproteinase-9(MMP-9). Methods and materials: Macrophages were cultured in the presence of 10% FBS. For the various MMP genes analysis, RT-PCR and real-time PCR were performed. In addition, zymography assay performed for the MMP activity. The phosphorylation assay did for the ERK1/2 and NF${\kappa}B$ activation, and luciferase promoter assay was for the NF${\kappa}B$ activity. Results: CpG DNA induced the mRNA expression of MMP-2, MMP-9, and MMP-13, but not of MMP-7, MMP-8, and MMP-12, in a time-dependent manner. Especially, the mRNA expression of MMP-9 was strongly induced by CpG DNA using real-time RT-PCR. The TLR9 inhibitor, chloroquine, suppressed CpG DNA-induced MMP-9 expression and its activity. Moreover, CpG DNA induced the phosphorylation of ERK and the inhibition of ERK by U0126 suppressed CpG DNA-induced MMP-9 expression and its activity. CpG DNA stimulated $I{\kappa}B-{\alpha}$ degradation and luciferase activity. In addition, pretreatment of SN-50, the inhibitor of NF${\kappa}B$, strongly blocked the CpG DNA-induced MMP-9 expression and activity. Conclusion: These observations suggest that CpG DNA may play important roles in the activation of macrophages by regulating the production of MMP-9 via the sequential TLR9-ERK-NF${\kappa}B$ signaling pathway.

• Toxicological Research
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• v.20 no.1
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• pp.37-42
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• 2004

This study was undertaken to investigate comparative anti-tumor activity of water extracts of Phellinus gilvus (PGE), Phellinus linteus (PLE), and Phellinus baumii (PBE) in vitro. The anti-tumor activity in the present study was evaluated by sulforhodamine B (SRB) and microtetrazolium (MTT) assay in terms of cell survival level. The tumor cells (sarcoma 180 and P388) were treated with PGE, PLE, and PBE (7.5, 15, and 30 $\mu\textrm{g}$/ml) and Doxorubicin (DOX) (0.001~10 $\mu\textrm{M}$). The results showed that DOX, PGE, and PLE inhibited proliferation showing a dose-dependent manner against both tumor cells. However, PBE was inhibited by the only 30 $\mu\textrm{g}$/ml in both cells proliferation. In conclusion, all of PGE, PLE, and PBE used in this study have shown anti-tumor activity against both sarcoma 180 and P388. Among them, PLE was the most effective in anti-tumor activity against sarcoma 180 (p<0.05) and PGE was against P388 in SRB assay. PLE, however, was against P388 (p<0.05) in MTT assay.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.1
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• pp.115-119
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• 2008

This study was done to evaluate the antioxidant effect of Crataegi Fructus (CF) extract on the oxidative stress induced by reactive oxygen species (ROS), The human skin fibroblasts (Detroit 551) were cultured with various concentrations of hydrogen peroxide $(H_2O_2)$. The cytotoxicity of $H_2O_2-induced$ oxidative stress was performed by XTT assay for the cell viability according to the dose- and time-dependent treatment. For the protective effect of CF extract on $H_2O_2-mediated$ oxidative stress, cell viability, lactate dehydroganase (LDH) activity, and ferric thiocyanate (FTC) assay for the inhibitive activity of lipid peroxidation on CF extract were carried out. In this study, $H_2O_2-mediated$ oxidative stress was decreased cell viability dose-, and time-dependent manner and increased LDH activity compared with the control in these cultures. In the protective effect, CF extract increased cell viability and decreased LDH activity on $H_2O_2-mediated$ oxidative stress, especially, CF extract has antioxidant effect by the showing the inhibitive activity of lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2-mediated$ oxidative stress was highly toxic, and also, CF extract showed the protective effect on $H_2O_2-mediated$ oxidative stress by showing the increased cell viability, decreased LDH activity and lipid peroxidation inhibition in these cultures.

• Korean Journal of Medicinal Crop Science
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• v.22 no.1
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• pp.8-16
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• 2014

The present study was carried out to evaluate the antioxidant and anti-inflammatory activities of three extracts (hot water, 50% ethanol and mixed solvent;water, ethanol, butylene glycol, propylene glycol) of dried chestnut, chestnut shell, chestnut leaves and dried chestnut leaves obtained from Castanea crenata tree. When conducted DPPH assay, radical scavenging activity of ethanol extract of chestnut shell was the highest with $IC_{50}$ $10.8{\mu}g/mL$ among four extracts from these parts (p < 0.05). In additional results by the xanthine oxidase assay, antioxidant activity showed that water extract of chestnut leaves showed the highest xanthine oxidase inhibitory activity in the tested extracts (p < 0.05). Futhermore, extracts of chestnut shell and leaves exhibited no cytotoxicity in RAW 264.7 cells (p < 0.05). Also, anti-inflammatory activity by NO assay showed LPS-induced NO was significantly inhibited following treatment with extracts of chestnut shell and leaves of 3mg/mL (p < 0.05). These data suggest that extract of chestnut shell have antioxidant and anti-inflamantory activity including chestnut leaves. Therefore, it is considered that Castanea crenata research range and selection of functional material can broaden chestnut shell to other fractions such as chestnut and chestnut leaves.

• YAKHAK HOEJI
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• v.52 no.5
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• pp.355-360
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• 2008

In this study we investigated antioxidant activity of against several free radicals and skin whitening effect of 70% ethanol extract (leaf extracts and branch/stem mixed) of Lindera obtusiloba BL. Antioxidant activity was assessed by DPPH, superoxide radical and hydroxyl radical assays. The Lindera obtusiloba BL. extract had antioxidant activity dose dependently with an ${IC}_{50}$ value of 243.14 and 181.10 ${\mu}g$/ml for DPPH, 165.77 and >1500 ${\mu}g$/ml for non-enzymatic system of superoxide radical assay, 35.47 and >100 ${\mu}g$/ml for enzymatic system of superoxide radical assay, 1.21 mg/ml for hydroxyl radical assay. In addition we tested tyrosinase inhibition activity and melanin contents on B16 melanoma F10. B16 melanoma cell was treated by such sample as 1, 5, 10 and 50 ${\mu}g$/ml for 72 hr and tyrosinase inhibition was tested. Melanogenesis was inhibited to 22% at the dose of 50 ${\mu}g$/ml and tyrosinase was inhibited to 45.2% at the same dose. In conclusion Lindera obtusiloba BL had potent antioxidant activity and inhibitory activity of tyrosinase and melanin formation. It could be developed as the health functional food and functional cosmetic resources.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2000.12a
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• pp.53-53
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• 2000

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.278-285
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• 1998

The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

• Clinical and Experimental Reproductive Medicine
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• v.40 no.1
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• pp.12-22
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• 2013

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.7 no.1
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• pp.109-116
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• 2001

By applying in vitro invasion assay model, we examined the anti-metatstastic effect of ChunghjagamTang(CLJGT). In 3H-thymidine incorporation assay, CLJGT treated groups showed the decreased DNA synthesis rate compared with control group. Gelatin zymogram assay showed that CLJGT decreases the gelatinolytic activity of MMP-9 from A-549, at the concentration of $800{\mu}g/ml$. We examined whether CLJGT inhibits the invasion of A-549 cells through the matrigel precoated transwell chamber. The results showed that CLJGT effectively inhibited the invasion of A-549 as compared with the control (+PMA) groups. From our research, part of mechanism underlying anti-metastastic effect of CLJGT was proven in vitro.

• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.171-175
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• 2006

Alnus japonica Steud (Betulaceae) has long been used as a Korean traditional medicine for gastric disorders, hepatitis, and fatty liver. As a part of our study on the identification of secondary metabolites of naturally occurring bioactive compounds, we isolated 1,7-bis(4-hydroxyphenyl)-3,5-heptanediol(1), 5-hydroxy-1,7bis(4-hydroxyphenyl)-3-heptanone(2), 5,3'-dihydroxy-7,4'-dimethoxyflavone(3) and 3,5,7,3',4'-pentahydroxyflavone(4) from the dichloromethane and ethylacetate-soluble fractions of Alnus japonica Steud. These compounds showed significant antioxidant activity in a concentration-dependent manner. The $IC_{50}$ values of compounds 1, 2, 3 and 4 were 30.1, 37.4, 20.2 and 13.7 ${\mu}g/mL$, respectively, through the scavenging capacity of 1,1-diphenyl-2-picrylhydrazyl radical assay.