• Title/Summary/Keyword: Gene expressions

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Effects of gangliosides from deer bone extract on the gene expressions of matrix metalloproteinases and collagen type II in interleukin-1β-induced osteoarthritic chondrocytes

  • Suh, Hyung Joo;Lee, Hyunji;Min, Byung Jung;Jung, Sung Ug;Jung, Eun Young
    • Nutrition Research and Practice
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    • v.10 no.6
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    • pp.569-574
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    • 2016
  • BACKGROUND/OBJECTIVES: We investigated the anti-osteoarthritic effects of deer bone extract on the gene expressions of matrix metalloproteinases (MMPs) and collagen type II (COL2) in interleukin-$1{\beta}$-induced osteoarthritis (OA) chondrocytes. MATERIALS/METHODS: Primary rabbit chondrocytes were treated as follows: CON (PBS treatment), NC (IL-$1{\beta}$ treatment), PC (IL-$1{\beta}+100{\mu}g/mL$ glucosamine sulphate/chondroitin sulphate mixture), and DB (IL-$1{\beta}+100{\mu}g/mL$ deer bone extract). RESULTS: The results of the cell viability assay indicated that deer bone extract at doses ranging from 100 to $500{\mu}g/mL$ inhibits cell death in chondrocytes induced by IL-$1{\beta}$. Deer bone extract was able to significantly recover the mRNA expression of COL2 that was down-regulated by IL-$1{\beta}$ (NC: 0.79 vs. DB: 0.87, P < 0.05) and significantly decrease the mRNA expression of MMP-3 (NC: 2.24 vs. DB: 1.75) and -13 (NC: 1.28 vs. DB: 0.89) in OA chondrocytes (P < 0.05).CONCLUSIONS: We concluded that deer bone extract induces accumulation of COL2 through the down-regulation of MMPs in IL-$1{\beta}$-induced OA chondrocytes. Our results suggest that deer bone extract, which contains various components related to OA, including chondroitin sulphate, may possess anti-osteoarthritic properties and be of value in inhibiting the pathogenesis of OA.

Involvement of Transglutaminase-2 in α-MSH-Induced Melanogenesis in SK-MEL-2 Human Melanoma Cells

  • Kim, Hyun Ji;Lee, Hye Ja;Park, Mi Kyung;Gang, Kyung Jin;Byun, Hyun Jung;Park, Jeong Ho;Kim, Mi Kyung;Kim, Soo Youl;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.3
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    • pp.207-212
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    • 2014
  • Skin hyperpigmentation is one of the most common skin disorders caused by abnormal melanogenesis. The mechanism and key factors at play are not fully understood. Previous reports have indicated that cystamine (CTM) inhibits melanin synthesis, though its molecular mechanism in melanogenesis remains unclear. In the present study, we investigated the effect of CTM on melanin production using ELISA reader and the expression of proteins involved in melanogenesis by Western blotting, and examined the involvement of transglutaminase-2 (Tgase-2) in SK-MEL-2 human melanoma cells by gene silencing. In the results, CTM dose-dependently suppressed melanin production and dendrite extension in a-MSH-induced melanogenesis of SK-MEL-2 human melanoma cells. CTM also suppressed a-MSH-induced chemotactic migration as well as the expressions of melanogenesis factors TRP-1, TRP-2 and MITF in a-MSH-treated SK-MEL-2 cells. Meanwhile, gene silencing of Tgase-2 suppressed dendrite extension and the expressions of TRP-1 and TRP-2 in a-MSH-treated SK-MEL-2 cells. Overall, these findings suggested that CTM suppresses a-MSH-induced melanogenesis via Tgase-2 inhibition and that therefore, Tgase-2 might be a new target in hyperpigmentation disorder therapy.

Specific Expression Patterns of xyl1, xyl2, and xyl3 in Response to Different Sugars in Pichia stipitis

  • Han, Ji-Hye;Park, Ju-Yong;Kang, Hyun-Woo;Choi, Gi-Wook;Chung, Bong-Woo;Min, Ji-Ho
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.946-949
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    • 2010
  • The effects of two different sugars (glucose and xylose) on the expression levels and patterns of the xylose reductase (xyl1), xylitol dehydrogenase (xyl2), and xylulokinase (xyl3) genes were analyzed using Pichia stipitis. A significant increase in mRNA levels of xyl1 was observed after 6 h growth in culture conditions using xylose as a sole carbon source, but expressions of the three genes were not influenced by normal culture media with glucose. In addition, expressions of xyl2 and xyl3 were not observed during the entire culture period during which xylose was added. It also was found that the expression level of xyl1 increased as a function of the xylose concentration (40, 60, and 80 g/l) used in this study, indicating that xyl1 expression sensitively responded to xylose in the culture media. Although the induced level of xyl2 increased slightly after 48 h in the xylose-supplemented culture conditions, the expression of xyl2 was not observed in the xylitol-supplemented culture conditions. Finally, considering the expression of each gene in response to glucose or xylose, the absolute expression levels of the three genes indicate that xyl1 is induced primarily by exposure to xylose.

Chemokines Gene Expression of RAW 264.7 Cells by Actinobacillus actinomycetemcomitans Lipopolysaccharide Using Microarray and RT-PCR Analysis

  • Chung, Jin;Choi, Mun Jeoung;Jeong, So Yeon;Oh, Jong Suk;Kim, Hyung Keun
    • Molecules and Cells
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    • v.27 no.2
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    • pp.257-261
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    • 2009
  • Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) is an important pathogen casuing aggressive periodontitis. The present study was designed to investigate the chemokines expression regulated by A. actinomycetemcomitans lipopolysaccharide (LPS). Chemokines genes expression profiling was performed in Raw 264.7 cells by analyses of microarray and reverse transcription-polymerase chain reaction (RT-PCR). Microarray results showed that the induction of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-$1{\alpha}$ (MIP-$1{\alpha}$), MIP-$1{\beta}$, MIP-$1{\gamma}$, regulated upon activation, normal T-cell expressed and secreted (RANTES), macrophage inflammatory protein-2 (MIP-2), and interferon-${\gamma}$ inducible protein 10 (IP 10) by A. actinomycetemcomitans LPS was increased to 12.5, 1.53, 9.09, 17.3, 2.82, 16.1, and 18.1 folds at 18 h, respectively. To check these chemokines expression by A. actinomycetemcomitans LPS, we examined gene expressions by RT-PCR, and found that the expression of MIP-$1{\beta}$, MIP-$1{\gamma}$, RANTES, MIP-2, and IP 10 was increased 107.1, 93.6, 106.8, 86.5, and 162.0 folds at 18 h, respectively. These results indicate that A. actinomycetemcomitans LPS stimulates the several chemokines expressions (MIP-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, RANTES, MIP-2, and IP 10) in Raw 264.7 cells.

Analysis of Endoplasmic Reticulum (ER) Stress Induced during Somatic Cell Nuclear Transfer (SCNT) Process in Porcine SCNT Embryos

  • Lee, Hwa-Yeon;Bae, Hyo-Kyung;Jung, Bae-Dong;Lee, Seunghyung;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Development and Reproduction
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    • v.22 no.1
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    • pp.73-83
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    • 2018
  • This study investigates the endoplasmic reticulum (ER) stress and subsequent apoptosis in duced during somatic cell nuclear transfer (SCNT) process of porcine SCNT embryos. Porcine SCNT and in vitro fertilization (IVF) embryos were sampled at 3 h and 20 h after SCNT or IVF and at the blastocyst stage for mRNA extraction. The x-box binding protein 1 (Xbp1) mRNA and the expressions of ER stress-associated genes were confirmed by RT-PCR or RT-qPCR. Apoptotic gene expression was analyzed by RT-PCR. Before commencing SCNT, somatic cells treated with tunicamycin (TM), an ER stress inducer, confirmed the splicing of Xbp1 mRNA and increased expressions of ER stress-associated genes. In all the embryonic stages, the SCNT embryos, when compared with the IVF embryos, showed slightly increased expression of spliced Xbp1 (Xbp1s) mRNA and significantly increased expression of ER stress-associated genes (p<0.05). In all stages, apoptotic gene expression was slightly higher in the SCNT embryos, but not significantly different from that of the IVF embryos except for the Bax/Bcl2L1 ratio in the 1-cell stage (p<0.05). The result of this study indicates that excessive ER stress can be induced by the SCNT process, which induce apoptosis of SCNT embryos.

A Vinegar-processed Ginseng Radix (Ginsam) Ameliorates Hyperglycemia and Dyslipidemia in C57BL/KsJ db/db Mice

  • Han, Eun-Jung;Park, Keum-Ju;Ko, Sung-Kwon;Chung, Sung-Hyun
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1228-1234
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    • 2008
  • Having idea to develop more effective anti-diabetic agent from ginseng root, we comprehensively assessed the anti-diabetic activity and mechanisms of ginsam in C57BL/KsJ db/db mice. The db/db mice were divided into 4 groups; diabetic control (DC), ginsam at a dose of 300 or 500 mg/kg (GS300 or GS500) and metformin at a dose of 300 mg/kg (MT300). Ginsam was orally administered for 8 weeks. GS500 reduced the blood glucose concentration and significantly decreased an insulin resistance index. In addition, GS500 reduced the plasma non-esterified fatty acid, triglyceride, and increased high density lipoprotein-cholesterol as well as decreased the hepatic cholesterol and triglyceride. More interestingly, ginsam increased the plasma adiponectin level by 17% compared to diabetic control group. Microarray, quantitative-polymerase chain reaction and enzyme activity results showed that gene and protein expressions associated with glycolysis, gluconeogenesis, and fatty acid oxidation were changed to the way of reducing hepatic glucose production, insulin resistance and enhancing fatty acid $\beta$-oxidation. Ginsam also increased the phosphorylation of AMP-activated protein kinase and glucose transporter expressions in the liver and skeletal muscle, respectively. These changes in gene expression were considered to be the mechanism by which the ginsam exerted the anti-diabetic and anti-dyslipidemic activities in C57BL/KsJ db/db mice.

St. John's Wort (Hypericum perforatum) stimulates human osteoblastic MG-63 cell proliferation and attenuates trabecular bone loss induced by ovariectomy

  • You, Mi-kyoung;Kim, Du-Woon;Jeong, Kyu-Shik;Bang, Mi-Ae;Kim, Hwan-Seon;Rhuy, Jin;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.9 no.5
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    • pp.459-465
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    • 2015
  • BACKGROUND/OBJECFTIVES: The effect of St. John's Wort extract (SJW) on MG-63 cell proliferation and trabecular bone loss induced by ovariectomy was examined. MATERIALS/METHODS: Proliferation, expression of estrogen receptor (ER) ${\alpha}$ and ER ${\beta}$, and gene expressions of osteoprotegerin (OPG), osteocalcin (OC) and alkaline phosphatase (ALP) were examined in MG-63 cells treated with or without SJW. Ovariectomized rats were treated with SJW at the dose of 100 or 200 mg/kg/day, ${\beta}$-estradiol-3-benzoate (E2), or vehicle only (OVX-C), and sham operated rats were treated with vehicle only (Sham-C). Serum ALP and C-telopeptide (CTX), and femoral trabecular bone loss were examined. RESULTS: SJW increased MG-63 cell proliferation and expression of ER ${\alpha}$ and ER ${\beta}$, and positive effect was shown on gene expressions of ALP, OC and OPG. SJW also showed estrogen like effect on bone associated with slowing down in trabecular bone loss. Histopathology by H&E showed rats treated with SJW displayed denser structure in metaphyseal region of distal femur compared with rats in OVX-C. SJW was shown to reduce serum CTX in OVX rats. CONCLUSION: The present study provides new insight in preventing estrogen deficiency induced bone loss of SJW and possibility for its application in bone health supplement.

Hsa_Circ_0001947/MiR-661/DOK7 Axis Restrains Non-Small Cell Lung Cancer Development

  • Bao, Yuyan;Yu, Yanjie;Hong, Bing;Lin, Zhenjian;Qi, Guoli;Zhou, Jie;Liu, Kaiping;Zhang, Xiaomin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1508-1518
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    • 2021
  • Hsa_circ_0001947 is associated with multiple cancers, but its function in non-small cell lung cancer (NSCLC) is ambiguous and needs further research. The targeting relationship among circ_0001947, miR-661, and downstream of tyrosine kinase 7 (DOK7) was predicted by database and further verified by dual-luciferase reporter assay, while their expressions in cancer tissues and cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). After transfection, cell biological behaviors and expressions of miRNAs, miR-661 and DOK7 were determined by cell function experiments and qRT-PCR, respectively. Circ_0001947 was low-expressed in NSCLC tissues and cells. Circ_0001947 knockdown intensified cell viability and proliferation, induced cell cycle arrest at S phase, suppressed apoptosis and evidently enhanced miR-510, miR-587, miR-661 and miR-942 levels, while circ_0001947 overexpression did the opposite. MiR-661 was a target gene of circ_0001947 that participated in the regulation of circ_0001947 on cell biological behaviors. Furthermore, DOK7, the target gene of miR-661, partly participated in the regulation of miR-661 on cell viability. Hsa_circ_0001947 acts as a sponge of miR-661 to repress NSCLC development by elevating the expression of DOK7.

Ethanol extract of Sinsun-yukza-hwan, a Korean medicinal prescription, promotes hair growth in C57BL/6 mice, an alopecia animal model

  • Kim, Ji Yoon;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.33 no.3
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    • pp.1-9
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    • 2018
  • Objectives : In Korean medicine, a prescription of Sinsun-yukza-hwan (Shenxian-liuzi-wan, SSY) has been used in clinic for treatment of alopecia via oral. This study was performed to determine transdermal effects of the ethanol extract from SSY on hair growth and -related gene expressions in mice. Methods : We analyzed index compound, 5-hydroxy-methyl-2-furaldehyde (HMF), in SSY extract by ultra performance liquid chromatography (UPLC). 6 weeks old C57BL/6 mice with removed hair were used as an alopecia animal model. Mice were divided into 3 experimental groups including normal (3 water: 1 ethanol: 2 polyethylene glycol mixture as a vehicle), SSY extract and 5% minoxidil (as a positive control), treated groups. SSY was applied topically on the hair-shaved skin of C57BL/6 mice every day for 15 days. The color, thickness and density of hair were monitored every $5^{th}$ day by naked eye, photograph and phototrichogram using folliscope. Also hair growth-associated gene expressions were measured by immunoblotting assay. Results : Hair density of minoxidil or SSY-treated group was significantly increased compared to that of vehicle application on the $15^{th}$ day, respectively. And hair thickness of minoxidil and SSY groups was increased compared to that of vehicle treated group on the $15^{th}$ day, respectively. Induction of insulin-like-growth factor 1(IGF-1) and vascular endothelial growth factor (VEGF) were also significantly accelerated by SSY extract compared to those of vehicle-applied group. Conclusions : These results provide scientific evidence to support the potent multi-application of SSY as a cosmeceutical material for promoting hair growth.

Comprehensive Lipid Profiling Recapitulates Enhanced Lipolysis and Fatty Acid Metabolism in Intimal Foamy Macrophages From Murine Atherosclerotic Aorta

  • Jae Won Seo;Kyu Seong Park;Gwang Bin Lee;Sang-eun Park;Jae-Hoon Choi;Myeong Hee Moon
    • IMMUNE NETWORK
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    • v.23 no.4
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    • pp.28.1-28.20
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    • 2023
  • Lipid accumulation in macrophages is a prominent phenomenon observed in atherosclerosis. Previously, intimal foamy macrophages (FM) showed decreased inflammatory gene expression compared to intimal non-foamy macrophages (NFM). Since reprogramming of lipid metabolism in macrophages affects immunological functions, lipid profiling of intimal macrophages appears to be important for understanding the phenotypic changes of macrophages in atherosclerotic lesions. While lipidomic analysis has been performed in atherosclerotic aortic tissues and cultured macrophages, direct lipid profiling has not been performed in primary aortic macrophages from atherosclerotic aortas. We utilized nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry to provide comprehensive lipid profiles of intimal non-foamy and foamy macrophages and adventitial macrophages from Ldlr-/- mouse aortas. We also analyzed the gene expression of each macrophage type related to lipid metabolism. FM showed increased levels of fatty acids, cholesterol esters, phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, and sphingomyelin. However, phosphatidylethanolamine, phosphatidic acid, and ceramide levels were decreased in FM compared to those in NFM. Interestingly, FM showed decreased triacylglycerol (TG) levels. Expressions of lipolysis-related genes including Pnpla2 and Lpl were markedly increased but expressions of Lpin2 and Dgat1 related to TG synthesis were decreased in FM. Analysis of transcriptome and lipidome data revealed differences in the regulation of each lipid metabolic pathway in aortic macrophages. These comprehensive lipidomic data could clarify the phenotypes of macrophages in the atherosclerotic aorta.