• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.938-943
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• 2002

This study was perfonned to detect polymorphisms of the two candidate genes, bovine BTN (Butyrophilin) and ST AT5a (Signal Transducers and Activators of Transcription) gene using 98 Holstein bulls' frozen semen, and to offer the basic information for QTL (Quantitative Trait Loci) analysis. Each BTN PCR product was digested with endonuclease restriction enzyme. The digested fragments of four BTN PCR products were observed as follows: 316,280, and 162 bp in BTN1, 568, 305 and 263 bp in BTN2, 576, 332, and 244 bp in BTN3, and 573, 291, and 282 bp in BTN4, respectively. The gene frequencies of A and B allele in four BTN loci were as follows: 0.8980 and 0.1020 in BTN1, 0.5510 and 0.4490 in BTN2, 0.8163 and 0.1837 in BTN3, and 0.8875 and 0.1122 in BTN4, respectively. And three genotypes (homotypel, heterotype, and homotype2) for STAT5a were observed by SSCP (single stranded conformational polymorphism) method and the genotype frequencies are 78.57%, 19.39%, and 2.04%, respectively. The PlC (Polymorphism Information Content) value and heterozygosity of four BTN loci were as follows: 0.1695 and 0.1870 in BTN1, 0.3713 and 0.4927 in BTN2, 0.2549 and 0.2999 in BTN3, and 0.1794 and 0.1992 in BTN4, respectively. Comparing with the reported data, PlC value of BTN2 might have the possibility to be useful marker. Other BTN loci indicated skewed allele distribution.

• Archives of Pharmacal Research
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• 제29권5호
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• pp.405-411
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• 2006

Biphenyl dimethyl dicarboxylate (DDB) is a hepatoprotectant, which is used as an adjuvant agent in a treatment for chronic hepatitis. Amantadine is an antiviral agent, which is utilized primarily in the treatment of influenza, but also, occasionally in the treatment of hepatitis C. In a previous study, we reported that DDB, coupled with amantadine, would exert an anti-HBV effect, via the induction of interferon-inducible gene expression in the HepG2 2.2.15 cell line. The primary objective of the present study was to determine whether or not DDB and/or amantadine exhibit anti-HBV properties, and what mechanisms of action might be involved in such properties. In our study, we were able to determine that DDB stimulates Jak/Stat signaling, and induces the expression of interferon alpha $(IFN-\alpha)$ stimulated genes, most notably 6-16 and ISG12. In addition, the antiviral effectors induced by $IFN-\alpha$, PKR, OAS, and MxA, were regulated in the presence of DDB at its optimal concentration $(250{\mu}g/mL)$, to a degree commensurate with the degree of induction associated with the $IFN-\alpha$ treated group. Finally, we determined that the replication of pregenomic RNA and HBeAg was inhibited by DDB treatment, and this inhibition was maximized when coupled with the administration of amantadine $(25{\mu}g/mL)$. In conclusion, the results of this study demonstrated clearly that DDB, as well as the combination of DDB/amantadine, directly inhibited $IFN-\alpha$ signaling-mediated replication of HBV in infected hepatocytes, and thus may represent a novel treatment for chronic hepatitis B, which would be characterized principally by its improved safety over other treatment strategies.

• 대한약침학회지
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• 제16권4호
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• pp.7-13
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• 2013

Objectives: Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored. Therefore, in this study, we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C (ethanolic extract of Gonolobus condurango diluted $10^{-60}$ times) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol (placebo) control. Methods: We checked the activity of different signal proteins (like $p21^{WAF}$, p53, Akt, STAT3) related to deacetylation, cell growth and differentiation by western blotting and analyzed cell-cycle arrest, if any, by fluorescence activated cell sorting. After viability assays had been performed with Condurango 30C and with a placebo, the activities of histone de-acetylase (HDAC) enzymes 1 and 2 were measured colorimetrically. Results: While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced HDAC2 activity quite strikingly, it apparently did not alter the HDAC1 enzyme; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity. Data on $p21^{WAF}$, p53, Akt, and STAT3 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and G1-phase cell-cycle arrest when Condurango 30C was used at a 2% dose. Conclusion: Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells, which the placebo was unable to do.

• The Korean Journal of Physiology and Pharmacology
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• 제26권6호
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• pp.457-468
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• 2022

It has been demonstrated that APPL1 (adaptor protein, phosphotyrosine interacting with PH domain and leucine zipper 1) is involved in the regulation of several growth-related signaling pathways and thus closely associated with the development and progression of some cancers. Diallyl trisulfide (DAT), a garlic-derived bioactive compound, exerts selective cytotoxicity to various human cancer cells through interfering with pro-survival signaling pathways. However, whether and how DAT affects survival of human hepatocellular carcinoma (HCC) cells remain unclear. Herein, we tested the hypothesis of the involvement of APPL1 in DAT-induced cytotoxicity in HCC HepG2 cells. We found that Lys 63 (K63)-linked polyubiquitination of APPL1 was significantly decreased whereas phosphorylation of APPL1 at serine residues remained unchanged in DAT-treated HepG2 cells. Compared with wild-type APPL1, overexpression of APPL1 K63R mutant dramatically increased cell apoptosis and mitigated cell survival, along with a reduction of phosphorylation of STAT3, Akt, and Erk1/2. In addition, DAT administration markedly reduced protein levels of intracellular TNF receptor-associated factor 6 (TRAF6). Genetic inhibition of TRAF6 decreased K63-linked polyubiquitination of APPL1. Moreover, the cytotoxicity impacts of DAT on HepG2 cells were greatly attenuated by overexpression of wild-type APPL1. Taken together, these results suggest that APPL1 polyubiquitination probably mediates the inhibitory effects of DAT on survival of HepG2 cells by modulating STAT3, Akt, and Erk1/2 pathways.

• 대한한방내과학회지
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• 제27권1호
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• pp.72-83
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• 2006

Objective : This study was designed to identify lipid protection formation in stratum corneum and anti-inflammatory effects of Sopungdojeok-tang(SD) on atopic dermatitis(AD). Materials and Methods : In Vivo, SD extract was orally administered to BALB/c mice at $2.5m{\ell}/kg/day$ for 2 days after 5% sodium dodecyl sulfate evoked atopic dermatitis in abdominal skin. Morphological changes were observed by immunohistochemical stain using monoclonal antibodies(BrdU, ceramide, MIP-2, $NF-{\kappa}B$ p50, IL-4, and STAT6) and TUNEL method. In vitro, the alterations of IL-4 mRNA expression were detected by RT-PCT in SD extract treated EL4 cells after phorbol-12-myristate-13-acetate and 4-tert-Octylphenol induce Th2 skewed condition. Results : SD is used in Oriental Medicine for its potential curative for atopic dermatitis. In this study, we have investigated the anti-inflammatory and lipid lamella repair effects of SD were investigated. SD decreased the number of eosinophil in atopic dermatitis induced mice. In the histological properties, the hyperplasia, edema, infiltration of lymphocytes, damage of intercellular space of stratum corneum, BrdU positive reacted cells in stratum basal, and degranulated mast cells and capillaries in dermal papillae decreased in mice with SD. Treatment of SD also decreased MIP-2, STAT6 and IL-4 in dermal papillae. The IL-4 mRNA expression decreased in a dose-dependant manner in SD treated EL4 cells. In addition, decrease of $NF-{\kappa}B$ p50 and increase of apoptotic cells in dermis were observed in SD treated mice. These data suggest that SD may beneficial for atopic dermatitis. Conclusions : These data suggest that SD is beneficial in treatment of atopic dermatitis, and that SD provides lipid protection in stratum corneum and anti-inflammatory effects on atopic dermatitis.

• 한국동물위생학회지
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• 제34권2호
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• pp.159-166
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• 2011

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

• 생명과학회지
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• 제21권3호
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• pp.349-357
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• 2011

EGFR kinase의 활성을 억제할 수 있는 억제제는 암뿐만이 아니라 성장성 질환에도 효과적인 치료제가 될 수 있다. 본 연구는 새로운 퀴나졸린계 물질인 화합물 63013과 63033의 EGFR kinase 활성억제 효과를 분석하였다. 이들 물질들은 기존의 디알콕시퀴나졸린의 용해성을 증가시키기 위하여 [1,4]-다이옥시노 퀴나졸린 구조를 가지며 알콕시 곁사슬로 연결되어있다. 화합물 63013과 63033은 A431 인간 피부암세포에서 EGF에의해 유도되는 EGFR의 kinase 활성을 저해, 세포 내에서 EGFR 신호체계에 관여하는 MEK1/2, MAPK p44/42, AKT, STAT3과 같은 하위 분자들의 활성저해 효과를 유도하였다. 이러한 활성저해 효과는 현재 상용화되어 있는 Gefitinib (Iressa$^{(R)}$)와의 비교연구에서 화합물 63013과 63033이 보다 더 낮은 처리 농도에서 EGFR kinase의 활성을 저해하며 암세포의 성장을 억제함을 관찰 할 수 있었다. 따라서 본 연구는 이들 신규 물질들의 EGFR-연관 질환에 대한 EGFR kinase 선택적 억제제로서의 이용 가능성을 제시하고 있다.

• Journal of Microbiology and Biotechnology
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• 제21권2호
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• pp.176-182
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• 2011

The study objective was to prepare biodegradable branched dextran microspheres encapsulated with His-tagged interferon-alpha (BDM-hIFN-${\alpha}$) and evaluate its activity in vitro and in vivo. The glycidyl methacrylate derivatized dextrans (Dex-GMA) as precursor was primarily synthesized by substituting hydroxyl groups of either the branched or linear type of dextran with GMA. Dex-GMA microspheres loaded with hIFN-${\alpha}$ was then prepared by the water-in-water emulsion technique. In vitro release and Western blotting experiments demonstrated the retained activity of hIFN-${\alpha}$ released from branched dextran microspheres at 24 h by inducing phosphorylation of signal transducer and activator transcription-1 (STAT-1), a down-stream effector of IFN-${\alpha}$, in HepG2 cells. Animal data further revealed a peak of plasma levels of IFN-${\alpha}$ in rats injected intravenously with BDM-hIFN-${\alpha}$ at 10 min post-injection, but a sharp decline at 2 h. High plasma levels of neopterin, a plasma protein induced by IFN-${\alpha}$, were also detected in rats injected with BDM-hIFN-${\alpha}$ at 10 min post-injection. Notably, plasma levels of neopterin remained high at 4 h, but largely declined thereafter.

• 대한한방내과학회지
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• 제38권4호
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• pp.468-478
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• 2017

Objectives: This study investigated the effects of Douchi Hataedock on Th2-skewed conditions to control allergic rhinitis. Methods: NC/Nga mice were divided into three groups: 10 mice were assigned to the control group (CTRL; no treatment), 10 mice to allergic rhinitis-induced (ARE) without treatment group, and 10 mice to the allergic rhinitis-induced (FGT) after Douchi Hataedock treatment group. The 3-week-old mice of the FGT group were given one 10 mg/kg dose of Douchi Hataedock extract and resensitized to allergic antigens at weeks four, five, and six. Allergic rhinitis was induced primarily in mice nasal cavities for five days after one week of final sensitization. The second induction used the same method one week after the first induction was completed. After one week, the nasal mucosal tissues of each group were observed. Immunohistochemical staining for IL-4, STAT6, CD40, $Fc{\varepsilon}RI$, substance P, MMP-9, $NF-{\kappa}B$ p65, p-IkB, and iNOS in the nasal mucosa was also performed. Results: The FGT group had less respiratory epithelial damage and less mucin secretion in goblet cells than the ARE group and showed a 62% decrease in IL-4, 85% decrease in STAT6, 71% decrease in CD40, 69% decrease in $Fc{\varepsilon}RI$, 43% decrease in substance P, 49% decrease in MMP-9, 43% decrease in NF-kB p65, 38% decrease in p-IkB, and 73% decrease in iNOS compared to the ARE group. Conclusions: Douchi Hataedock lessens inflammation in epithelial and goblet cells and reduces inflammatory mediator secretion in a mouse allergic rhinitis model.

• The Korean Journal of Physiology and Pharmacology
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• 제25권3호
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• pp.207-216
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• 2021

Several studies have previously reported that exposure to stress provokes behavioral changes, including antinociception, in rodents. In the present study, we studied the effect of acute cold-water (4℃) swimming stress (CWSS) on nociception and the possible changes in several signal molecules in male ICR mice. Here, we show that 3 min of CWSS was sufficient to produce antinociception in tail-flick, hot-plate, von-Frey, writhing, and formalin-induced pain models. Significantly, CWSS strongly reduced nociceptive behavior in the first phase, but not in the second phase, of the formalin-induced pain model. We further examined some signal molecules' expressions in the dorsal root ganglia (DRG) and spinal cord to delineate the possible molecular mechanism involved in the antinociceptive effect under CWSS. CWSS reduced p-ERK, p-AMPKα1, p-AMPKα2, p-Tyk2, and p-STAT3 expression both in the spinal cord and DRG. However, the phosphorylation of mTOR was activated after CWSS in the spinal cord and DRG. Moreover, p-JNK and p-CREB activation were significantly increased by CWSS in the spinal cord, whereas CWSS alleviated JNK and CREB phosphorylation levels in DRG. Our results suggest that the antinociception induced by CWSS may be mediated by several molecules, such as ERK, JNK, CREB, AMPKα1, AMPKα2, mTOR, Tyk2, and STAT3 located in the spinal cord and DRG.