• 대한한방내과학회지
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• 제20권1호
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• pp.99-110
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• 1999

Effects of Chiyangtang(CYT) on H. pylori-induced increase of interleukin 8 and interleukin 1 gene expression was studied in Kato Ⅲ cell line, a human stomach epithelial cell line. Treatment of H. pylori to the cell culture signifant!y increased IL-8 and IL-1 mRNA synthesis. When CYT was added along with H. pylori, the increase of IL-8 and IL-1 mRNA synthesis was blocked. Activation of transcription factor $NF-{\kappa}B$ and AP-1 which were known to important in IL-8 and IL-1 gene expression was also studied using chloramphenicol acetyltransferase(CAT) assay. Treatment of H. pylori increased activation of $NF-{\kappa}B$ and AP-l and CYT effectively protected the activation. Electrophoretic mobility shift assay suggested that CYT effectively inhibited DNA binding of $NF-{\kappa}B$ and AP-l to their cognate site. These results suggested that CYT could prevent stomach diseases through the down regulation of IL -8 and IL-l gene expression which might be mediated by the inhibition of $NF-{\kappa}B$ and AP-1 activities and their binding to DNA.

• Animal cells and systems
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• 제15권3호
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• pp.219-225
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• 2011

We studied oxidative stress in cinnamon clownfish exposed to hypoosmotic (35 psu ${\rightarrow}$ 17.5 psu and 17.5 psu with prolactin (PRL)) and low temperature ($28^{\circ}C{\rightarrow}24^{\circ}C$ and $20^{\circ}C$) conditions by measuring the expression and activity of Cu/Zn-superoxide dismutase (Cu/Zn-SOD), catalase (CAT), and glutathione peroxidase (GPX). The expression and activity of the antioxidant enzymes were significantly higher after the fish were exposed to $24^{\circ}C$, $20^{\circ}C$, and 17.5 psu, and expression was repressed by PRL treatment. Furthermore, we measured $H_2O_2$ and lipid peroxidation levels and found that they were significantly higher after exposure to the hypoosmotic and low-temperature environments. Additionally, we investigated changes in plasma AST and ALT levels after exposure to low temperature and hypoosmotic stress. These levels increased upon exposure of the clownfish to $24^{\circ}C$, $20^{\circ}C$, and 17.5 psu, but the levels of these parameters decreased in the 17.5 psu with PRL treatment during a salinity change. The results indicate that hypoosmotic and low-temperature conditions induce oxidative stress in cinnamon clownfish and that the parameters tested in this study may be indices of oxidative stress in the cinnamon clownfish.

• Archives of Pharmacal Research
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• 제28권3호
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• pp.351-357
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• 2005

Endosulfan is one of the organochlorine pesticides, which are well-known endocrine disruptors (EDs), and it acts as an estrogen agonist. Estrogen is a group of hormones that play an important role in mammary gland function and are implicated in mammary carcinogenesis. In the present study, we studied the effects of endosulfan on nodule like alveolar lesion (NLAL) formation in mouse mammary gland development using a mouse mammary gland organ culture (MMOC) system. Although endosulfan-treated mammary glands did not form NLALs, more alveolar buds were formed in this group than in the negative control (vehicle-treated) group. In addition, telomerase reverse transcriptase (TERT) mRNA expression levels were increased in endosulfan-treated mammary glands in a dose-dependent manner. Telomerase can be activated by estrogen, therefore, we examined the effects of endosulfan on telomerase activity, and found that the telomerase activity in estrogen receptor-positive MCF-7 cells was up-regulated by endosulfan treatment. Moreover, this activation was accompanied by the up­regulation of the TERT mRNA expression. Also, transient expression assays using CAT reporter plasm ids containing various fragments of the TERT promoter showed that this imperfect palindromic estrogen-responsive element is almost certainly responsible for the transcriptional activation by endosulfan. These results may help elucidate the endocrine disrupting mechanism of endosulfan.

• The Korean Journal of Physiology and Pharmacology
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• 제5권6호
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• pp.521-527
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• 2001

Diclofenac, a phenylacetic acid derivative, is a widely used non-steroidal anti-inflammatory drug (NSAID) to provide effective relief of inflammation and pain. Nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation. We examined the inhibitory effects of diclofenac on the induction of iNOS in RAW 264.7 macrophages which were activated with lipopolysaccharide (LPS) plus interferon-gamma $(IFN-{\gamma}).$ Treatment of RAW 264.7 cells with diclofenac and other NSAIDs (aspirin and indomethacin) significantly inhibited NO production and iNOS protein expression induced by LPS plus $IFN-{\gamma}.$ Also, diclofenac but not aspirin and indomethacin, inhibited iNOS mRNA expression and nuclear factor-kappa B $(NF-{\kappa}B)$ binding activity concentration-dependently. Furthermore, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that only diclofenac inhibited the iNOS promoter activity induced by LPS plus $IFN-{\gamma}$ through the $NF-{\kappa}B$ sites of iNOS promoter. Taken together, these suggest that diclofenac may exert its anti-inflammatory effect by inhibiting iNOS gene expression at the transcriptional level through suppression of $NF-{\kappa}B$ activation.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.277-286
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• 2016

Makgeolli lees (ML) has several physiological effects such as antioxidant, antidiabetic, and anticancer properties, but its biological functions have not been determined definitively. Here, we tested whether ML has a cytoprotective effect on paraquat (PQ)-induced oxidative stress in the human lung carcinoma cell line A549. At 0.1 mg/ml ML, viability of PQ-exposed A549 cells was restored by 12.4%, 18.5%, and 48.6% after 24, 48, and 72 h, respectively. ML also reduced production of the intracellular reactive oxygen species (ROS) that were generated by PQ treatment. Further experiments revealed that ML treatment enhanced the expression and nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) as well as ARE-GFP reporter activity. ML treatment also effectively increased the expression of NRF2's target genes NAD(P)H dehydrogenase quinone 1 (NQO1) and heme oxygenase 1 (HO-1). Moreover, we found that expression of cytoprotective genes, including glutathione peroxidases (GPXs), superoxide dismutase (SOD1), catalase (CAT), peroxiredoxin 3 (PRDX3), and peroxiredoxin 4 (PRDX4), was greatly enhanced by treatment with ML during PQ exposure. Taken together, the data suggest that treatment of PQ-exposed A549 cells with ML ameliorates cytotoxicity through induction of NRF2 expression and its target genes HO-1, NQO1, and other antioxidant genes. Thus, ML may serve as a functional food applicable to ROS-mediated human diseases.

• Mycobiology
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• 제46권4호
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• pp.382-387
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• 2018

The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

• 한국미생물·생명공학회지
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• 제25권6호
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• pp.571-579
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• 1997

For the overproduction of pectate lyase (PL), the recombinant plasmid pl2BS fl which has strong promoter from alkali-tolerent Bacillus sp. YA-14 was used. In order to overexpress the pectate lyase by the action of overlapping strong promoter in pl2BS$\Delta$fl, 1.6 kb of PL gene was inserted into pl2BS$\Delta$fl to form pl2BS$\delta$f1-PL and the enzyme was expressed. But decreased expression efficiency of the PL gene was observed and it was due to the presence of the transcription terminator region on the upstream of the PL gene. The transcription terminator of the PL gene in pl2BS$\delta$f1-PL was removed and the resulting plasmid p12BS$\Delta$fl$\Delta$PL was formed. Bacillus subtilis 207-25 harboring the recombinant plasmid, p12BS$\Delta$fl$\Delta$PL, revealed increased expression efficiency with chloramphenicol induction when cat-86 was used as a reporter gene.

• Bulletin of the Korean Chemical Society
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• 제26권9호
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• pp.1385-1389
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• 2005

Human hepatitis B virus (HBV) is a pathogen related to the development of liver diseases including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). However, the efficient methods to suppress HBV replication have not been developed yet. Therefore, we have used RNA interference (RNAi) as a potential tool for the suppression of HBV replication. Here, we designed a 21 nt small intefering dsRNA (siRNA) against hepatitis B virus X (HBx) RNA with 3' overhanging ends derived from T7 promoter. It has been reported that HBV X protein plays an important role in HBV gene expression and viral replication. The suppression of HBx gene expression by the 21 nt siRNA was investigated by Northern blot analysis and chloramphenicol acetyl transferase (CAT) assay. The level of HBx mRNA was decreased by siRNA in a dose-dependent manner. We also found that the 21 nt siRNA inhibited the HBV replication in hepatocellular carcinoma cell.

• BMB Reports
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• 제44권12호
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• pp.805-810
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• 2011

Methionine sulfoxide reductase A (MSRA) is a ubiquitous enzyme that has been demonstrated to reduce the S enantiomer of methionine sulfoxide (MetSO) to methionine (Met) and can protect cells against oxidative damage. In this study, we isolated a novel MSRA (SlMSRA2) from Micro-Tom (Solanum lycopersicum L. cv. Micro-Tom) and characterized it by subcloning the coding sequence into a pET expression system. Purified recombinant protein was assayed by HPLC after expression and refolding. This analysis revealed the absolute specificity for methionine-S-sulfoxide and the enzyme was able to convert both free and protein-bound MetSO to Met in the presence of DTT. In addition, the optimal pH, appropriate temperature, and $K_m$ and $K_{cat}$ values for MSRA2 were observed as 8.5, $25^{\circ}C$, $352{\pm}25\;{\mu}M$, and $0.066{\pm}0.009\;S^{-1}$, respectively. Disk inhibition and growth rate assays indicated that SlMSRA2 may play an essential function in protecting E. coli against oxidative damage.

• 대한수의학회지
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• 제39권4호
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• pp.772-777
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• 1999

The uncaped genomic RNA of bovine viral diarrhea virus (BVDV) initiates translation by recruitment of eukaryotic translation initiation factors at the internal ribosome entry site (IRES). N-terminal protease ($N^{pro}$) is the first translation product of the open reading frame (ORF). By using the vaccinia virus SP6 RNA polymerase transient expression system, we showed previously that deletion of $N^{pro}$ region reduced translation by 21%. To better understand the biological significance of $N^{pro}$ for translation, we carried out a mutational analysis of the $N^{pro}$ region of BVDV cloned in the intercistronic region of a bicistronic reporter plasmid. We constructed a bicistronic expression vector in which the entire 5 UTR and the mutated $N^{pro}$ region (${\Delta}386-901$, ${\Delta}415-901$ and ${\Delta}657-901$) was cloned between two reporter genes, chloramphenicol acetyltransferase (CAT) and luciferase (LUC). In vivo translation analyses showed that $N^{pro}$ region was dispensible for efficient translation. The results indicate that the $N^{pro}$ region is not essential for BVDV RNA translation and the 3' boundary of BVDV IRES is expanded into $N^{pro}$ region, suggesting that $N^{pro}$ may not play a major role in BVDV replication.