• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.42 no.4
• /
• pp.403-412
• /
• 2016

The tight junction, one of Intercellular junctions, performs a variety of biological functions by bonding adjacent cells, including the barrier function to control the movement of the electrolyte and water. Recent studies have revealed that unusual expression of tight junction-related genes have been shown to be related in cancer development and progression. Recently, there are many reports that control of tight junction proteins expression is closely related to the skin moisture. In this study, we are focusing on the regulating mechanism of tight junction-associated genes by the steviol and its derivatives. Steviol, used as a sweetner, is known to chemical compound isolated from stevia plant. The MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay was carried out in HaCaT cells (human keratinocyte cell line) in order to determine the cytotoxicity. As a result, while steviol showing cytotoxicity from $250{\mu}M$, steviol derivatives are not cytotoxic more than $250{\mu}M$ concentration. We have observed a change in the tight junction protein via quantitative real-time PCR. Claudin 8 among tight junction proteins is only significantly reduced up to 30% in the presence of steviol. In addition, cell migration was inhibited by steviol, not by stevioside and rebaudioside. Finally, we could observe that steviol, not stevioside and rebaudioside, is able to increase the skin barrier permeability through the transepithelial electric resistance (TEER) measurements. These results suggest that the steviol and its derivatives are specifically acts on the tight junction related gene expression, but steviol derivatives are more suitable as a cosmetic material.

• Development and Reproduction
• /
• v.11 no.1
• /
• pp.49-54
• /
• 2007

Ethane 1,2-dimethane sulfonate(EDS), a toxin which specifically kills Leydig cells(LC), has been widely used to prepare the reversible testosterone(T) depletion rat model. In the present study, we monitored the gene expression profiles of pituitary gonadotropins, LH and FSH, up to 7 weeks after EDS injection. Adult male Sprague-Dawley rats($300{\sim}350\;g$ B.W.) were injected with a single dose of EDS(75 mg/kg i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. Total RNAs were purified from each pituitary, and the message levels of common alpha subunit($C{\alpha}$) of pituitary glycoprotein hormones, LH beta subunit($LH{\beta}$), FSH beta subunit($FSH{\beta}$) and GnRH receptor(GnRH-R) were evaluated by semi-quantitative RT-PCRs. The message levels of $C{\alpha}$ increased sharply during weeks 1-4, then return to the control level on week 5. The mRNA levels of $LH{\beta}$ were elevated after week 2, reached the peak at week 4, then declined to the control level after week 5. The message levels of $FSH{\beta}$ were elevated after week 2, reached the peak at week 3, then declined to the nadir at week 5. Similarly, the mRNA levels of GnRH-R were elevated after week 2, reached the peak at week 3, then gradually declined to the control level after week 5. The present study indicated that EDS treatment could induce reversible alterations in the transcriptional activities of gonadotropin subunits and GnRH-R in the anterior pituitary from male rats. EDS injection model might be useful to understand the mechanism of hormonal regulation of hypothalamus- pituitary neuroendocrine axis in male rats.

• Journal of Life Science
• /
• v.22 no.10
• /
• pp.1359-1370
• /
• 2012

Arabidopsis AtERF71/HRE2, a transcription activator, is located in the nucleus and is involved in the signal transduction of low oxygen and osmotic stresses. In this study, microarray analysis using AtERF71/HRE2-overexpressing transgenic plants was performed to identify genes downstream of AtERF71/HRE2. A total of 161 different genes as well as AtERF71/HRE2 showed more than a twofold higher expression in AtERF71/HRE2-overexpressing transgenic plants compared with wild-type plants. Among the 161 genes, 24 genes were transcriptional regulators, such as transcription factors and DNA-binding proteins, based on gene ontology annotations, suggesting that AtERF71/HRE2 is an upstream transcription factor that regulates the activities of various downstream genes via these transcription regulators. RT-PCR analysis of 15 genes selected out of the 161 genes showed higher expression in AtERF71/HRE2-overexpressing transgenic plants, validating the microarray data. On the basis of Genevestigator database analysis, 51 genes among the 161 genes were highly expressed under low oxygen and/or osmotic stresses. RT-PCR analysis showed that the expression levels of three genes among the selected 15 genes increased under low oxygen stress and another three genes increased under high salt stress, suggesting that these genes might be downstream genes of AtERF71/HRE2 in low oxygen or high salt stress signal transduction. Microarray analysis results indicated that AtERF71/HRE2 might also be involved in the responses to other abiotic stresses and also in the regulation of plant developmental processes.

• Journal of Applied Biological Chemistry
• /
• v.64 no.4
• /
• pp.421-431
• /
• 2021

It has been suggested that some toothpastes have the potential to promote hair growth. However, there was no scientific verification on the hair growth effect of toothpaste and no scientific report on major active ingredients in toothpaste. In this work, toothpaste and its constituents were applied topically over the shaved skin of C57BL/6 mice and evaluated. Results indicated that toothpaste showed hair growth effect. Also, the effect of toothpaste constituents on the proliferation rate of keratinocyte cells was investigated. The mixture solution of 𝛼-tocopherol acetate, l-menthol, and stevioside, each of that was known to promote hair growth and other toothpaste constituents were applied topically on mouse skin. When the mixture solution was included, hair growth effect was observed in mice. Transcriptome analysis was performed using the dorsal epidermis of mice from the group treated with toothpaste, the mixture which are presumed to be active ingredients for hair growth, and from mice used for the control group. As a result of analyzing the genes whose expression was significantly changed in each treatment group, the gene patterns of the two groups were very similar. Also, when functional genomic analysis was performed, genes with functions related to hair growth regulation showed a high extent of the change in both groups. Hair growth-related genes whose expression was changed in both groups included keratin, keratin-related proteins, forkhead box, and sonic hedgehog. Therefore, the hair growth effect of toothpaste is thought to be due to the effect of a mixture of 𝛼-tocopherol acetate, l-menthol, and stevioside.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.39 no.2
• /
• pp.193-202
• /
• 2010

In vitro activities of Codonopsis lanceolata (CL) 70% ethanol extract and its fractions (hexane, chloroform, ethyl acetate, butanol and water) were examined by total polyphenol content, reducing power, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS), 2,2-diphenyl-$\beta$-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity (ORAC) assays. The ethyl acetate fraction from CL ethanol extract (CLEA) showed the highest total polyphenol content (22.7 mg/g) among five fractions, and also exhibited an excellent reducing power (0.42~1.27 at $250\sim1,000\;{\mu}g/mL$). CLEA at $100\sim400\;{\mu}g/mL$ concentrations had 27.7~70.3% of ABTS radical scavenging activity and the highest DPPH radical scavenging activity (81.6% at $400\;{\mu}g/mL$). CLEA had dominantly higher $ORAC_{{ROO}{\cdot}}$activity compared to other fractions. CLEA and butanol fraction had significantly higher $ORAC_{{OH}{\cdot}}$ activities than 70% ethanol extract, hexane, chloroform and water fractions. The CLEA exhibited the highest antioxidant activity in CL 70% ethanol extract and its fractions. Thus, effect of CLEA treatment on antioxidant gene expression under the oxidative stress conditions by a high fat diet in animal model was studied by microarray and RT-PCR methods. The 31 antioxidant genes were expressed but the genes were not up-regulated at least a two-fold by CLEA treatment. We concluded that CLEA does not have an indirect antioxidant effect but a direct antioxidant effect by up-regulation of antioxidant genes in high fat diet-induced obese mice.

• Food Science and Preservation
• /
• v.18 no.3
• /
• pp.366-373
• /
• 2011

Recent studies suggested that Cheonnyuncho is a significant source of bioactive phenolic compounds, comparable to phytochemicals, including green tea and onion. In this study, the hot-water and 80% ethanolic extracts of Cheonnyuncho were assessed as to their total phenol content, total flavonoids content, antioxidant activity (DPPH radical-scavenging activity and reducing power), and anti-obesity activity. The results showed that the total phenol contents of the hot water extract and the 80% ethanolic extract were $16.52{\pm}3.87$ and $13.44{\pm}0.85$ mg GAE/g, respectively. The total flavonoids content was detected only in the 80% ethanolic extract, however, with a 778.08 ${\mu}g$ catechin equivalents/g content. The DPPH radical-scavenging activity and reducing power of the 80% ethanolic extract from Cheonnyuncho was significantly higher than those of the water extract (p < 0.05). During the adipocyte differentiation, the 80% ethanolic extract of Cheonnyuncho more significantly inhibited lipid accumulation and ROS production than the 3T3-L1 cells that were treated with hot water extract. Furthermore, the 80% ethanolic extract of Cheonnyuncho suppressed the mRNA abundance of the adipogenic transcription factor, $PPAR{\gamma}$ (peroxisome proliferator-activated receptor ${\gamma}$), and its target gene, aP2 (adipocyte protein 2). These results indicate that Cheonnyuncho extracts can inhibit adipogenesis through a mechanism that involves direct down regulation of $PPAR{\gamma}$ gene expression or via modulation of ROS production associated with radical-scavenging activities.

• Molecules and Cells
• /
• v.37 no.2
• /
• pp.178-186
• /
• 2014

Differential transcription of the clusterin (CLU) gene yields two CLU isoforms, a nuclear form (nCLU) and a secretory form (sCLU), which play crucial roles in prostate tumorigenesis. Pro-apoptotic nCLU and anti-apoptotic sCLU have opposite effects and are differentially expressed in normal and cancer cells; however, their regulatory mechanisms at the transcriptional level are not yet known. Here, we examined the transcriptional regulation of nCLU in response to hypoxia. We identified three putative hypoxia response elements (HREs) in the human CLU promoter between positions -806 and +51 bp. Using a luciferase reporter, electrophoretic gel mobility shift, and chromatin immunoprecipitation assays, we further showed that hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) bound directly to these sites and activated transcription. Exposure to the hypoxia-mimetic compound $CoCl_2$, incubation under 1% $O_2$ conditions, or overexpression of HIF-$1{\alpha}$ enhanced nCLU expression and induced apoptosis in human prostate cancer PC3M cells. However, LNCaP prostate cancer cells were resistant to hypoxia-induced cell death. Methylation-specific PCR analysis revealed that the CLU promoter in PC3M cells was not methylated; in contrast, the CLU promoter in LNCap cells was methylated. Co-treatment of LNCaP cells with $CoCl_2$ and a demethylating agent promoted apoptotic cell death through the induction of nCLU. We conclude that nCLU expression is regulated by direct binding of HIF-$1{\alpha}$ to HRE sites and is epigenetically controlled by methylation of its promoter region.

• Journal of Life Science
• /
• v.26 no.9
• /
• pp.991-998
• /
• 2016

NF-κB acts as a critical transcription factor for the survival of cells via the induction of antiapoptotic genes. Constitutive activation of NF-κB in many types of solid tumors suggests that the inhibition of NF-κB might prevent or inhibit tumorigenesis. Although a number of studies demonstrated that Hsp70 regulated NF-κB activity, the exact mechanism is not clear. This study investigated the functional relationship of Hsp70 and IKKγ in the regulation of NF-κB activation using expression plasmids of components of the IKK complex. Wild-type and deletion mutants of IKKγ were expressed together with Hsp70, and the combined regulatory effect of Hsp70 and IKKγ on NF-κB activation was assayed. Hsp70 suppressed the activation of NF-κB in a reporter plasmid assay. Hsp70 also suppressed the phosphorylation and degradation of IκBα. The suppressive effect of Hsp70 on NF-κB activation was synergistically elevated by IKKγ. The N-terminal IKKβ binding site, C-terminal leucine zipper, and zinc finger domains of IKKγ were not necessary for the suppressive effect. Furthermore, Hsp70 and IKKγ synergistically suppressed the induction of COX-2 expression by lipopolysaccharides in RAW264.7 cells. These results suggest that overexpression of Hsp70 and IKKγ may be a strategic method for inhibition of NF-κB and related diseases.

• Food Science and Preservation
• /
• v.21 no.2
• /
• pp.264-275
• /
• 2014

This study evaluated the improving efficacy of Lespedeza cuneata ethanol extract on skin photoaging induced by ultraviolet (UV) irradiation. The total polyphenol and flavonoid contents of the extract were respectively $134.98{\pm}1.70$ and $16.20{\pm}0.05$ mg/g, respectively. The superoxide anion radical scavenging activity and electron-donating ability of the extract were shown to be dependent on concentration, and the antioxidant ability was shown to be more effective in superoxide anion radical scavenging activity than in electron-donating ability under the same concentration conditions. In the in vivo test conducted using hairless mouse with skin photoaging induced by UVB irradiation, the skin erythema of the groups treated with the extract (AS) reduced to 28% of the control, and the skin moisture content increased to 131%.. The extract treatment of the UV-damaged skin improved the morphological and histopathological state of the skin. Furthermore, the SOD, GST and CAT activities in the skin tissue of the AS group increased, and the XO activity and TBARS generation decreased. With regard to the genes related to the photoaging skin, the expression of PAK, p38, c-Fos, c-Jun, TNF-${\alpha}$ and MMP-3 in the skin of the AS group were found to have decreased. It was therefore concluded that Lespedeza cuneata ethanol extract can reduce wrinkle formation in the skin due to the regulation of the gene expression caused by the exposure to UVB light.

• Korean Journal of Environmental Biology
• /
• v.39 no.3
• /
• pp.354-361
• /
• 2021

Understanding the phytotoxic mechanism of methyl bromide (MB), an essential fumigant during the quarantine and pre-shipment process, is urgently needed to ensure its proper use and reduce international economic losses. In a previous study, two main MB-induced toxic mechanisms such as reactive oxygen species (ROS) and auxin distribution were selected by analyzing transcriptomic analysis. In the study, a 3-week-old A. thaliana was supplied with 1 mM ROS scavengers [N-acetyl-L-cysteine (NAC) or L-glutathione (GSH)] and 1µM indole-3-acetic acid(IAA) three times every 12 h, and visual and gene expression assessments were performed to evaluate the reduction in phytotoxicity by supplements. Phytotoxic effects on the MB-4h exposed group were decreased with GSH application compared to the other single supplements and a combination of supplements at 7 days post fumigation. Among these supplements, GSH at a concentration of 1, 2, and 5mM was suppled to A. thaliana with MB-fumigation. During a long-term observation of 2 weeks after the fumigation, 5 mM GSH application was the most effective in minimizing MB-induced phytotoxic effects with up-regulation of HSP70 expression and increase in main stem length. These results indicated that ROS was a main key factor of MB-induced phytotoxicity and that GSH can be used as a supplement to reduce the phytotoxicity of MB.