• Journal of Genetic Medicine
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• v.4 no.2
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• pp.179-185
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• 2007

Purpose : Catecholamines are the neuro-transmitters in the sympathetic nervous system (SNS) and are activated by stress stimulus. Tyrosine hydroxylase (TH) and Dopamine-${\beta}$-Hydroxylase (DBH) are very important enzymes in the catecholamine synthesis. Corticotropin releasing hormone (CRH) is released in the process of reacting to stresses. The aim of this study is to find out what effects immobilization stresses have on the expression of TH, BDH and CRH mRNA in a rat's brains. Methods : We compare expression levels in rat's brains of TH, DBH and CRH mRNA induced by immobilization stresses between the test group and controled group. The expression levels of TH, DBH and CRH mRNA are measured by RT-PCR and the Western Blotting Analysis (WBA). Results : In brains and adrenal glands of the immobilization stress group, the expression levels of TH and DBH mRNAs are significantly two to three times higher (P<0.01), and CRH mRNAs are approximately one and a half times higher (P<0.05) than those of controlled group. Conclusion : This study suggest that the expression levels of TH, DBH and CRH mRNAs are activated by stress stimulus in a rat's brains and adrenal glands.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.109-113
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• 2001

As the complete genomic sequences accumulate, the use of global techniques became possible. DNA microarray is a powerful technology for measuring global mRNA levels. This method, however, does not provide information on post-translational modifications of proteins. In addition, mRNA levels do not strictly correlate with protein concentrations, especially for lower-abundance proteins. Therefore, studies at the level of transcription are not sufficient to understand cellular activity. Proteomic techniques to analyze protein expression and function at the large-scale have been developed and used. This review introduces a simple explanation for proteomic analysis and examples of how proteomics is applied in cell biology.

• The Korean Journal of Zoology
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• v.33 no.4
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• pp.435-445
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• 1990

Gonadotropin releasing horrnone (GnRH) is known to be extrahypothalamically localized with a broad range including gonad. It remains, however, unknown whether GnRH is locally synthesized in the gonad. The present srudy aims to identity expression and cellular localization of GnRH-Iike mRNA and immunoreactive GnRH in the rat gonad. GnRH radioimmunoassay and chromatographic extracts on G-50 sephadex column showed that rat gonadal extracts contained a substantial amount of immunoreactive GnRH similar to the hypothalamic and synthetic GnRH. Although a wide distribution of immunostainable GnRH-like molecule with different cell types in the rat ovary was observed, the major cell population hybridized with GnRH probe appears to be granulosa. theca cells and corpus luteum. Immunoreactive GnRH-Iike peptides were distributed m various regions of testis, including spermatogenic cells, Sertoli cells and Leydig cells. In situ hybridization revealed that positive signals of GnRH-Iike mRNA were predominandy present in Sertoli cells within some seminiferous tubules, but absent in the outside of seminiferous tubules in the testis. This study clearly demonstrated that GnRH-Iike molecule present in the rat gonad may be resulted from the local synthetic machinery of GnRH supporting the notion that this peptide may act as autocrine and/or paracrine role in intra-gonadal communication.

• Journal of Life Science
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• v.30 no.6
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• pp.501-512
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• 2020

Sasa quelpaertensis Nakai leaf has been used as a folk medicine for the treatment of gastric ulcer, dipsosis, and hematemesis based on its anti-inflammatory, antipyretic, and diuretic characteristics. We have previously reported the procedure for deriving a phytochemical-rich extract (PRE) from S. quelpaertensis and how PRE and its ethyl acetate fraction (EPRE) exhibits an anticancer effect by inducing apoptosis in various gastric cancer cells. To explore the molecular targets involved in this apoptosis, we investigated the mRNA and microRNA profiles of EPRE-treated SNU-16 human gastric cancer cells. In total, 2,875 differentially expressed genes were identified by RNA sequencing, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that the EPRE-modulated genes are associated with apoptosis, mitogen-activated protein kinase, inflammatory response, tumor necrosis factor signaling, and cancer pathways. Subsequently, protein-protein interaction network analysis confirmed interactions among genes associated with cell death and apoptosis, and 27 differentially expressed microRNAs were identified by further sequencing. Here, GO and KEGG pathway analysis revealed that EPRE modified the expression of microRNAs associated with the cell cycle and cell death, as well as signaling of tropomyosin-receptor-kinase receptor, transforming growth factor-b, nuclear factor kB, and cancer pathways. Taken together, these results provide insight into the mechanisms underlying the anticancer effect of EPRE.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.80-84
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• 2008

We constructed the null mutants of fission yeast Schizosaccharomyces pombe spDbp5 gene that is homologous to DEAD-box RNA helicase DBP5 in budding yeast Saccharomyces cerevisiae, which plays important roles in mRNA export out of nucleus. A null mutant in an $h^+/h^+$ diploid strain was constructed by replacing the spDbp5-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spDbp5 is essential for vegetative growth. The haploid spDbp5 null mutants harboring pREP81X-spDbp5 plasmid showed extensive $poly(A)^+$ RNA accumulation in the nucleus and decrease in the cytoplasm after repression of spDbp5 expression. These results suggest that spDbp5 is also involved in mRNA export from the nucleus.

• Tuberculosis and Respiratory Diseases
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• v.42 no.6
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• pp.862-870
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• 1995

Background: Oxygen free radicals have generally been considered as cytotoxic agents. On the other hand, recent results suggest that small nontoxic amounts of these radicals may act a role in intracellular signal transduction pathway and many efforts to reveal the role of these radicals as secondary messengers have been made. It is evident that the oxygen radicals are released by various cell types in response to extracellular stimuli including LPS, TNF, IL-1 and phorbol esters, all of which translocate the transcription factor $NF{\kappa}B$ from cytoplasm to nucleus by releasing an inhibitory protein subunit, $I{\kappa}B$. Activation of $NF{\kappa}B$ is mimicked by exposure to mild oxidant stress, and inhibited by agents that remove oxygen radicals. It means the cytoplasmic form of the inducible tanscription factor $NF{\kappa}B$ might provide a physiologically important target for oxygen radicals. At the same time, it is well known that LPS induces the release of oxygen radicals in neutrophil with the activation of $NF{\kappa}B$. From above facts, we can assume the expression of IL-8 and IL-$1{\beta}$ gene by LPS stimulation may occur through the activation of $NF{\kappa}B$, which is mediated through the release of $I{\kappa}B$ by increasing amounts of oxygen radicals. But definitive evidence is lacking about the role of oxygen free radicals in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. We conducted a study to determine whether oxygen radicals act a role in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. Method: Human peripheral blood monocytes were isolated from healthy volunteers. Time and dose relationship of $H_2O_2$-induced IL-8 and IL-$1{\beta}$ mRNA expression was observed by Northern blot analysis. To evaluate the role of oxygen radicals in the expression of IL-8 and IL-$1{\beta}$ mRNA by LPS stimulation, pretreatment of various antioxiants including PDTC, TMTU, NAC, ME, Desferrioxamine were done and Northern blot analysis for IL-8 and IL-$1{\beta}$ mRNA was performed. Results: In PBMC, dose and time dependent expression of IL-8 and IL-$1{\beta}$ mRNA by exogenous $H_2O_2$ was not observed. But various antioxidants suppressed the expression of LPS-induced IL-8 and IL-$1{\beta}$ mRNA expression of PBMC and the suppressive activity was most prominant when the pretreatment was done with TMTU. Conclusion: Oxygen free radical may have some role in the expression of IL-8 and IL-$1{\beta}$ mRNA of PBMC but that radical might not be $H_2O_2$.

• Development and Reproduction
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• v.4 no.1
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• pp.125-131
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• 2000

This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

• Journal of Veterinary Clinics
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• v.32 no.4
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• pp.289-294
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• 2015

Fucoidan which is sulfated polysaccharide extracted from brown seaweed has a wide variety of internal biological activities. The objectives of this study were to examine the effect of fucoidan on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to investigate whether this effect is involved in the expression of inducible nitric oxide synthase (iNOS) and the activation of activator portein-1 (AP-1). The levels of NO production and AP-1 activity in the culture supernatants from porcine PBMCs were measured by the enzyme-linked immunosorbent assay and the levels of iNOS and AP-1 mRNA were determined by real time polymerase chain reaction. Fucoidan in LPS-naïve PBMCs has no effects on the production of NO and activity of AP-1. Expressions of iNOS and AP-1 mRNA in LPS-naïve PBMCs were also not affected by treatment of fucoidan. However, NO production, AP-1 activity and expressions of iNOS and AP-1 mRNA were dramatically increased in PBMCs stimulated with LPS. Enhancing effects of NO production and AP-1 activity in PBMCs induced by LPS were reduced by addition of fucoidan. Fucoidan also inhibited an increase in expressions of iNOS and AP-1 mRNA in LPS-stimulated PBMCs. These results suggested that fucoidan exerts anti-inflammatory effect by down-regulating production of NO via suppressing expression of iNOS and activity of AP-1 in LPS-stimulated porcine PBMCs.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.622-629
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• 2014

We studied the effects of Akebia quinata fruit extract (AQ) on acute alcohol-induced hepatotoxicity in mice. AQ (30-1,000 mg/kg body weight (BW) per day) was orally administered to the study group, once daily for 1 week. On the last day of AQ treatment, ethanol (6 mg/kg BW) was orally administered to induce acute liver injury. The AQ-treated group showed significantly lower levels of alanine aminotransferase and aspartate aminotransferase, compared to the only ethanol-treated group (ETG). The glutathione level in the AQ-treated group elevated up to 20.6%, compared to that observed in the ETG. The mRNA expression of glutathione synthetic enzymes was also higher in the AQ-treated group, compared to the ETG. The AQ-treated group also exhibited lower levels of expression of NADPH oxidase 4 and tumor necrosis factor alpha mRNA. Thus, these results show that AQ treatment can be a potential method to reduce oxidative stress and inflammation in ethanol-treated mouse liver and also that AQ can be a useful therapeutic agent for acute alcohol-induced hepatotoxicity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.1
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• pp.57-67
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• 2019

Particulate matters with a diameter of < $10{\mu}m$ (PM10) exert oxidative stress and inflammatory events in various organs. The purpose of this study was to examine the molecular mechanism of reactive oxygen species (ROS) production induced by PM10 in the human epidermal keratinocytes (HEKs). When cultured HEKs were exposed to PM10, ROS production was induced and it was inhibited by apocynin, an antioxidant. The mRNA expression of NADPH oxidase (NOX) family was analyzed in order to examine their role in PM10-induced ROS production. PM10 increased the mRNA expression of NOX1, NOX2, dual oxidase (DUOX) 1 and DUOX2. HEKs expressed DUOX1 and DUOX2 at higher levels compared to other NOXs. The mRNA expression of dual oxidase maturation factors, DUOXA1 and DUOXA2, was also increased by PM10. We examined whether these calcium-dependent enzymes, DUOX1 and DUOX2, mediate the PM10-induced ROS production. A selective intracellular calcium chelator, BAPTA-AM, attenuated ROS production induced by PM10 or calcium ionophore A23187. The small intereference RNA (siRNA)-mediated down-regulation of DUOX2, but not DUOX1, attenuated the ROS production induced by PM10. PM10 increased the expression of inflammatory cytokines such as interleukin $(IL)-1{\beta}$, IL-6, IL-8 and interferon $(IFN)-{\gamma}$. SiRNA-mediated down-regulation of DUOX2 suppressed the PM10-induced expression of $IFN-{\gamma}$ but not other cytokines. This study suggests that DUOX2 plays a crucial role in ROS production and inflammatory response in PM10-exposed keratinocytes.