• BMB Reports
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• v.49 no.5
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• pp.270-275
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• 2016

The Ubiquitin proteasome system (UPS) plays roles in protein degradation, cell cycle control, and growth and inflammatory cell signaling. Dysfunction of UPS in cardiac diseases has been seen in many studies. Cholesterol acts as an inducer of cardiac hypertrophy. In this study, the effect of proteasome inhibitors on the cholesterol-induced hypertrophic growth in H9c2 cells is examined in order to observe whether UPS is involved in cardiac hypertrophy. The treatment of proteasome inhibitors MG132 and Bortezomib markedly reduced cellular surface area and mRNA expression of β-MHC in cholesterol-induced cardiac hypertrophy. In addition, activated AKT and ERK were significantly attenuated by MG132 and Bortezomib in cholesterol-induced cardiac hypertrophy. We demonstrated that cholesterol-induced cardiac hypertrophy was suppressed by proteasome inhibitors. Thus, regulatory mechanism of cholesterol-induced cardiac hypertrophy by proteasome inhibitors may provide a new therapeutic strategy to prevent the progression of heart failure.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.3
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• pp.225-231
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• 2014

In this study we aim to extensively investigate the anti-influenza virus immune responses in human pharyngeal epithelial cell line (Hep-2) and evaluate the protective role of Toll-like receptor (TLR) ligands in seasonal influenza A H1N1 (sH1N1) infections in vitro. We first investigated the expression of the TLRs and cytokines genes in resting and sH1N1 infected Hep-2 cells. Clear expressions of TLR3, TLR9, interleukin (IL)-6, tumour necrosis factor (TNF)-${\alpha}$ and interferon (IFN)-${\beta}$ were detected in resting Hep-2 cells. After sH1N1 infection, a ten-fold of TLR3 and TLR9 were elicited. Concomitant with the TLRs activation, transcriptional expression of IL-6, TNF-${\alpha}$ and IFN-${\beta}$ were significantly induced in sH1N1-infected cells. Pre-treatment of cells with poly I:C (an analog of viral double-stranded RNA) and CpG-ODN (a CpG-motif containing oligodeoxydinucleotide) resulted in a strong reduction of viral and cytokines mRNA expression. The results presented indicated the innate immune response activation in Hep-2 cells and affirm the antiviral role of Poly I:C and CpG-ODN in the protection against seasonal influenza A viruses.

• Archives of Pharmacal Research
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• v.28 no.12
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• pp.1358-1364
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• 2005

In this study, we investigated whether a novel anti-ischemic $K_{ATP}$ opener KR-31378 [(2S,3S,4R)­N'-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2 -methly-2-dimethoxymethly-2H-benzopyran-4-yl)­N'-benzylguanidine] has protective effect against oxidative stress-induced death in heart-derived H9c2 cells. Cell death was induced by BSO, butionine sulfoximine, which inhibits GSH synthesis and subsequently increases reactive oxygen species (ROS) level. Cell death was quantitatively determined by measuring lactate dehydrogenase (LDH) activity and stained by Hoechst 33258. BSO-induced ROS production and mitochondrial membrane potential (MMP) were measured using 2',7'-dichlorofluorescein diacetate oxidation and rhodamine 123, respectively. Both the LDH release and the ROS elevation induced by treatment of H9c2 cells with 10 mM BSO, were significantly decreased by KR-31378. These protective effect and antioxidant effect of KR-31378 appeared to be independent on $K_{ATP}$ channel opening. Cells exposed to BSO showed an early reduction in MMP, and this reduction in MMP was significantly reversed by treatment with KR-31378. Caspase-3 activity in BSO treated H9c2 cells was remarkably increased, and this increased caspase-3 activity was significantly reversed by KR-31378. In conclusion, our results suggest that KR-31378 can produce cardioprotective effect against oxidative stress-induced cell death through antioxidant mechanism.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.137-145
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• 2020

Oxidative stress is one of the pathogenic mechanisms of various neurodegenerative diseases, such as Alzheimer's disease. Neuroglia, the most abundant cells in the brain, is thought to play an important role in the antioxidant defense system and neuronal metabolic support against neurotoxicity and oxidative stress. We investigated the protective effect of paeoniflorin (PF) against oxidative stress in C6 glial cells. Exposure of C6 glial cells to hydrogen peroxide (H₂O₂, 500 μM) significantly decreased cell viability and increased amounts of lactate dehydrogenase (LDH) release, indicating H₂O₂-induced cellular damage. However, treatment with PF significantly attenuated H₂O₂-induced cell death as shown by increased cell survival and decreased LDH release. The H₂O₂-stimulated reactive oxygen species production was also suppressed, and it may be associated with improvement of superoxide dismutase activity by treatment with PF. In addition, an increase in ratio of Bcl-2/Bax protein expression was observed after treatment with PF. In particular, the down-stream of the apoptotic signaling pathway was inhibited in the presence of PF, mostly by reduction of cleaved-poly ADP ribose polymerase, cleaved caspase-3, and -9 protein expression. Furthermore, H₂O₂-induced phosphorylation of c-Jun N-terminal kinase and extracellular signal-regulated kinase 1/2 was attenuated by treatment with PF. Taken together, neuroprotective effect of PF against oxidative stress probably result from the regulation of apoptotic pathway in C6 glial cells. In conclusion, our findings suggest that PF may be a potent therapeutic agent for neurodegenerative disorders.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.9
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• pp.1242-1248
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• 2005

This study was designed to investigate the in vitro developmental ability and apoptosis of bovine embryos nucleartransferred (NT) with frozen-thawed or cooled donor cells. Cultured adult bovine ear cells were used as donor cells after sub-culturing to confluence (CC), cooling to 4$^{\circ}C$ for 48 h, or freezing-thawing (FT). Apoptotic cells in blastocysts were evaluated for apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Fusion, cleavage and blastocyst rates were 69.0 (167/242), 68.8 (115/167), and 29.9 (50/167) with CC cells, 70.4 (88/125), 69.3 (61/88), and 29.6 (26/88) with cooled cells and 66.1 (117/177), 70.1 (82/117), and 13.7 (16/117) with FT cells, respectively. Blastocyst rates of NT embryos derived from FT cells were significantly lower than those from CC or cooled cells (p<0.05). In addition, NT blastocysts produced by using FT cells showed significantly higher apoptosis rates (6.4${\pm}$4.0%) than those produced by CC (2.8${\pm}$1.7%) or cooled (2.3${\pm}$1.3%) cells. However, cooling of donor cells had no significant adverse effect on blastocyst rate as well as apoptosis rate. Therefore, our results suggest that cooled cells may be used as an alternative to freshly cultured confluent culture cells, as donor cells, for the production of Somatic nuclear cloned cattle.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.333-335
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• 1999

This study was carried out to find out the effect on fertilizing capacity according to sperm concentration of liquid boar semen. Four different doses with various motile sperm cells of 3.0$\times$10$^{9}$ , 2.5$\times$10$^{9}$ , 2.0$\times$10$^{9}$ , and $1.5\times$10$^{9}$ per 80$m\ell$ plastic bottle were inseminated twice 12 h interval after standing estrus in 6,818 sows. Farrowing rate and total piglets per litter were 82.2% and 10.9, respectively, with no significant differences among the other treatments. The presumption of optimal concentration of motile sperm cells in the liquid boar semen was best at 2.0~2.3$\times$10$^{9}$ per dose.

• The Korea Journal of Herbology
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• v.32 no.6
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• pp.9-15
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• 2017

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

• Korean Journal of Food Science and Technology
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• v.9 no.3
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• pp.183-189
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• 1977

Studies were made of the continuous production of Pyridoxal 5'-phosphate (pyridoxal-p) on simultaneously immobilized cell column. Whole-cell of Pseudomonas polycolor having high activity of pyridoxine 5'-phosphate (pyridoxine-p) oxidase and Kloeckera sp. No. 2201 having high activity of catalase were used as the enzyme materials. The enzyme sources were entrapped in a polyacrylamide gel. Enzymatic properties of the simultaneously immobilized cells were investigated, comparing with those of the mixed whole-cells of the microorganisms. The simultaneously immobilized cells had higher enzyme activity than singly immobilized cells of Pseudomonas polycolor. From this result, the simultaneously immobilized pyridoxine-p oxidase-catalase system could be available to exert a protective effect upon the pyridoxine-p oxidase by destroying $H_2O_2$ which is a by-product of pyridoxine-p oxidation. The optimum pH was 9.0 for the immobilized cells and the whole-cells. The optimum temperature was $45^{\circ}C$ for the immobilized cells and $40^{\circ}C$ for the whole-cells. The pyridoxine-p oxidase of the immobilized cells were activated by $Hg^{++}$ and some SH-compounds.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.644-649
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• 1998

To investigate the fermentation characteristics of Kimchi which was made at $12{\pm}1^{\circ}C$ and stored at $17{\pm}1^{\circ}C$ and $4{\pm}1^{\circ}C$, pH, total acid, total microbes, lactic acid bacteria cell count, dissolved $CO_2$content, reducing sugars and temperature at the center of Kimchi jar were measured and to know how much Japanese like to have Korean traditional Kimchi, Kimchi samples with 1.0, 2.0 and 3.0% salt contents were prepared and surved to panelist at $4{\pm}1^{\circ}C$. Sensory evaluation of Kimchi in terms of appearance, texture, carbonated mouthfeel, salty taste, sour taste, peppery taste and overall acceptability was done by scoring system with maximum 7 points and was analyzed statistically by SAS program. The results of fermentation of Kimchi which was made at $12{\pm}1^{\circ}C$ and stored at $17{\pm}1^{\circ}C\;and\;4{\pm}1^{\circ}C$ were as follows: pH and total acid content of Kimchi which was stored at $17{\pm}1^{\circ}C$ for 4 days were almost same as those of Kimchi at $4{\pm}1^{\circ}C$ for 48 days. Cell counts of total microbes and lactic acid bacteria in Kimchi which was stored at $17{\pm}1^{\circ}C$ for 2 days and at $4{\pm}1^{\circ}C\;for\;9\;days\;were\;1.5{\times}10^9\;cells/mL\;and\;6.3{\times}10^8\;cells/mL,\;2.0{\times}10^8\;cells/mL\;and\;8.7{\times}10^7\;cells/mL$, respectively. Time and temperature at the center of Kimchi jar during fermentation was took 25 and 35 hours to get to $17^{\circ}C\;and\;4^{\circ}C$ from initial temperature, respectively. Japanese who had experiences in eating Kimchi liked Korean traditional Kimchi with 2.0% of salt content. Sensory evaluation of Japanese consumer on peppery taste and overall acceptability of Kimchi showed significant difference (P<0.05).

• Biomedical Science Letters
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• v.12 no.3
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• pp.131-137
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• 2006

RGS is a negative regulator of G-protein signaling and can be identified by the presence of a conserved $120{sim}125$ amino acid motif, which is referred to as the RGS box. A number of RGSs are induced in response to a wide variety of stimuli. Increased levels of RGSs lead to significant decreases in GPCR responsiveness. To obtain further evidence of a role of RGS proteins in rat C6 astrocytoma cells, we first determined the expression profile of RGS-specific mRNA in C6 cells using reverse transcription-polymerase chain reaction (RT-PCR) with a poly dT18 primer and transcript-specific primers. We found that RGS2, RGS3, RGS6, RGS9, RGS10, RGS12, and RGS16 were differentially expressed in C6 astrocytoma cells. The highest expression rate was found for RGS3, followed by RGS16, RGS10 and RGS9, whereas the expression level for RGS2 was barely detectable. We next assessed whether forskolin regulated the expression of RGSs expressed in C6 astrocytoma cells. The present study found that forskolin dose-dependently stimulated the expression of RGS2 transcripts. This up-regulation of RGS2 gene was abrogated by H-89, potent and broad-spectrum protein kinase A (PKA) inhibitors. Actinomycin D completely inhibited the up-regulation of RGS2 gene induced by forskolin $(10{\mu}M)$, indicating that the regulation of RGS2 gene is controlled at the transcriptional level. In addition, forskolin did significantly activate transcriptional cAMP response element (CRE) in either HEK 293 cells or C6 cells and did not modulate the $NF-{\kappa}B$ and AP-l activity as measured by luciferase reporter gene assay. Finally, forskolin induced the expression of RGS2 mRNA in C6 astrocytoma cells, which depend on the PKA pathway and CRE transcriptional pathways.