• Proceedings of the PSK Conference
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• 2002.10a
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• pp.242.1-242.1
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• 2002

H$_2$O$_2$ has been shown to act as a signaling molecule involved in many cellular functions such as oxidant-induced stress, apoptosis, proliferation. In this study, we investigated the action mechanisms of H$_2$O$_2$ on activation of Extracellular Signal-Regulated Protein Kinase(ERK) in cultured feline ileal smooth muscle cells(ISMC). Western blot analysis done with phospho-specific MAP kinases antibodies demonstrated that potent activation of ERK and moderate activation of SAPK/JNK occurred within 30 min of H$_2$O$_2$ treatment. (omitted)

• Journal of Life Science
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• v.20 no.10
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• pp.1443-1450
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• 2010

Honokiol is a small molecular weight ligand originally isolated from the Chinese medicinal herb Magnolia officinalis, a plant used in traditional Chinese and Japanese medicine [9]. In a previous study, the effects of honokiol were shown to have anti-angiogenic, anti-invasive and anti-proliferative activities in a variety of cancers [1,3,4,11,13,17,24,29,30]. We showed previously that direct production of nitric oxide (NO) by treatment of NO donor, sodium nitroprusside (SNP), led to apoptosis in rabbit articular chondrocytes [15,16]. This study confirmed that NO-induced apoptosis was suppressed by honokiol treatment in a dose-dependent manner as determined by cell phenotype, MTT assay, Western blot analysis and FACS analysis in articular chondrocytes. Treatment of honokiol inhibited SNP-induced expression of p53 as well as DNA fragmentation in articular chondrocytes, but increased expressionof pro-caspase-3. Inhibition of SNP-induced apoptosis by honokiol treatment was rescued by LY294002, the specific inhibitors of phosphoinositide 3-kinase (PI-3K) in articular chondrocytes. Our results indicate that honokiol inhibits NO-induced apoptosis via PI-3K/AKT pathway in rabbit articular chondrocytes.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.62-66
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• 2001

This study was performed to investigate antigenic expression patterns in the course of HSV-1 infection. In SDS-PAGE analysis, HSV-1 antigens were detected, and among them, antigens in the size of 39, 47, 63, 86, 101, 105, 135, 159, and 181 kDa appear to be expressed in the most dominant forms. BALB/c mice were infected with HSV-1 for 29 days and antigenic expression from HSV-1 was investigated by Western blot analysis using anti-HSV-1 sera collected every two days from BALB/c mice infected with HSV-1. Most of HSV-1 antigens appeared sporadically as the infection progressed. However, antigens in the sizes of 63kDa and 135kDa were expressed from day 1 and 3, respectively, and existed continuously during the course of infection for 29 days, suggesting that they are the most dominant antigens inducing immune response durign HSV-1 infection, and they could be the target antigens for the development of vaccines. The isotype levels of IgA, IgGl, and IgM increased till the 17 th day infection and then started to decrease. During this course. IgGl was the most dominant isotype. In an indirect immunofluorescent assay, antibodies exhibited surface binding to the Vero cell infected with HSV-1, demonstrating that HSV-1 antigens are expressed on the surface of Vero cells.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.1-12
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Leonurus sibiricus on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Leonurus sibiricus and investigated cell death rate by MTS assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis and then we observed the differential gene expression by western blot analysis. Results : Leonurus sibiricus significantly inhibited the proliferation of uterine leiomyoma cell in a dose-dependent and time dependent manner. Fluorescence activated cell sorter (FACS) analysis indicated that Leonurus sibiricus induced G1 cell cycle arrest. Leonurus sibiricus enhanced the expression of p27 and p53 with cell cycle arrest. Conclusion : These findings suggest that Leonurus sibiricus is a candidate agent for the treatment of uterine leiomyoma. p27, $p53^{1}$ may play an important role in Leonurus sibiricus-induced cell cycle arrest and cell growth inhibition.

• The Korean Journal of Nuclear Medicine
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• v.37 no.6
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• pp.382-392
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• 2003

Purpose: Uptakes of Tc-99m MIBI (MIBI) and Tc-99m tetrofosmin (tetrofosmin) in human non-small cell lung cancer A549, multidrug-resistance associated protein (MRP) expressing cell, were investigated in vitro and in vivo. Materials and Methods: Western blot analysis and immunohistochemistry were used for detection of MRP in A549 cells with anti-MRPr1 antibody. Cellular uptakes of two tracers were evaluated at $100{\mu}M$ of verapamil (Vrp), $50{\mu}M$ of cyclosporin A (CsA) and $25{\mu}M$ of butoxysulfoximide (BSO) after incubation with MIBI and tetrofosmin for 30 and 50 min at $37^{\circ}C$, using single cell suspensions at $1{\times}10^6cells/ml$. Radioactivities in supernatants and pellets were measured with gamma well counter. A549 cells were inoculated in each flanks of 24 nude mice. Group 1 (Gr1) and Gr3 mice were treated with only MIBI or tetrofosmin, and Gr2 and Gr4 mice were treated with 70mg/kg of CsA i.p. for 1 hour before injection of 370KBq of MIBI or tetrofosmin. Mice were sacrificed at 10, 60 and 240 min. Radioactivities of organs and tumors were expressed as percentage injected dose per gram of tissue (%ID/gm). Results: Western blot analysis of the A549 cells detected expression of MRPr1 (190 kDa) and immunohistochemical staining of tumor tissue for MRPr1 revealed brownish staining in cell membrane but not P-gp. Upon incubating A549 cells for 60 min with MIBI and tetrofosmin, cellular uptake of MIBI was higher than that of tetrofosmin. Coincubation with modulators resulted in an increase in cellular uptakes of MIBI and tetrofosmin. Percentage increase of MIBI was higher than that of tetrofosmin with Vrp by 623% and 427%, CsA by 753% and 629% and BSO by 219% and 140%, respectively. There was no significant difference in tumoral uptakes of MIBI and tetrofosmin between Gr1 and Gr3. Percentage increases in MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively higher by the time up to 240 min with CsA. Conclusion: These results indicate that MIBI and tetrofosmin are suitable tracers for imaging MRP-mediated drug resistance in A549 tumors. MIBI may be a better tracer than tetrofosmin for evaluating MRP reversal effect of modulators.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1109-1121
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• 2009

In an attempt to discover novel biomarker proteins in type 2 diabetes prognosis, we investigated the influence of hypoglycemic extracellular polysaccharides (EPS) obtained from the macrofungus Tremella fuciformis on the differential levels of plasma proteins in ob/ob mice using two-dimensional gel electrophoresis (2-DE). The 2-DE analysis demonstrated that 92 spots from about 900 visualized spots were differentially regulated, of which 40 spots were identified as principal diabetes-associated proteins. By comparing control with EPS-fed mice, we found that at least six proteins were significantly altered in ob/ob mice, including Apo A-I, IV, C-III, E, retinol-binding protein 4, and transferrin, and their levels were interestingly normalized after EPS treatment. Western blot analysis revealed that the altered levels of the two regulatory molecules highlighted in diabetes and obesity (e.g., resistin and adiponectin) were also normalized in response to EPS. The Mouse Diabetes PCR Array profiles showed that the expression of 84 genes related to the onset, development, and progression of diabetes were significantly downregulated in liver, adipocyte, and muscle of ob/ob mice. EPS might act as a potent regulator of gene expression for a wide variety of genes in ob/ob mice, particularly in obesity, insulin resistance, and complications from diabetes mellitus.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.273-279
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• 2010

Follicular cystic follicles (FCFs) show delayed regression with persistent follicle growth. However, the mechanism by which follicles are persistently grown remains unclear. Glutathione S-transferases (GSTs) are drug-metabolizing and detoxification enzymes that are involved in the intracellular transport and metabolism of steroid hormones. In this study, a proteomic analysis was performed to identify whether GST expression is changed in bovine FCFs and to predict the interactions between GST and other proteins. Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and 25 mm). In bovine follicles, GST mu 1 (GSTM1) was detected as a differentially expressed protein (DEP) and significantly up-regulated in FCFs compared to normal follicles (p<0.05). Consistent with the proteomic results, semi-quantitative PCR data and western blot analysis revealed an up-regulation of GSTM1 in FCFs. Expression levels of aromatase and dehydrogenase proteins were changed in FCFs. These results show that the up-regulation of GSTM1 that is observed in bovine FCFs is likely to be responsible for the persistent follicle growth in FCFs as the activity of aromatase and the dehydrogenases.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.2
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• pp.25-42
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• 2007

Purpose : Uterine leiomyoma (fibroids) are benign smooth muscle tumors originating from the myometrium. These benign neoplasms of monoclonal origin are typically diagnosed during the reproductive years, occurring only after puberty and tending to regress after menopause. In the present study we used Chiljehyangbu-hwan to determine its growth inhibitory effect and apoptosis in human uterine leiomyoma cells. Methods : Primary cultured human uterine leiomyoma cells were treated with Chiljehyangbu-hwan. Cell viability analysis was analyzed by MTS assay and FACS was performed to ascertain the effects Chiljehyangbu-hwan. DNA fragmentation analysis and casapase-3 activity test were done. Expression of apoptosis related proteins were evaluated by Western blot analysis. Results : Cell viability was significantly influenced by Chiljehyangbu-hwan treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. FACS showed that Chiljehyangbu-hwan induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increased expression in Bid and Bax were observed. Chiljehyangbu-hwan treatment of uterine leiomyoma cells resulted in a concentration-dependent cell death induced via the mitochondrial pathway.

• The Korean Journal of Food And Nutrition
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• v.36 no.4
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• pp.296-308
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• 2023

This study investigated the anti-obesity effects of Coriandrum sativum L. ethanol extracts in a high fat diet-induced obesity model (DIO). We confirmed the anti-obesity effects by analysing the expression of the related proteins, weight gain, dietary intake, dietary efficiency, blood biochemistry, histological analysis and western blot analysis. After oral administration of Coriandrum sativumL. ethanol extracts at concentrations of 250 and 500 mg/kg, a significant improvement in dietary efficiency, reduction in weight gain, triglycerides, total cholesterol and LDL-cholesterol in blood lipid was observed for 8 weeks. In addition, improvement in blood glucose and metabolism confirmed through glucose tolerance test was observed. Further, the concentration of alanine transaminase (ALT) in blood was significantly decreased, which improved the fatty liver caused by high-fat diet intake as confirmed by liver tissue analysis. This phenomenon was confirmed to decrease the expression of fat accumulation-related PPARγ and FAS protein in the liver tissue. Especially, it is believed that FAS, a liposynthetic enzyme, has a stronger inhibitory effect than PPARγ. Therefore, Coriandrum sativum L. ethanol extract is thought to improve obesity by reducing blood lipids levels, improving glucose metabolism and inhibiting synthesis of the fat that accumulates in the liver in high-fat diet-induced obesity animal models.