• Title/Summary/Keyword: Expression and secretion

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Pear pomace ethanol extract improves insulin resistance through enhancement of insulin signaling pathway without lipid accumulation

  • You, Mi-Kyoung;Kim, Hwa-Jin;Rhyu, Jin;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.198-205
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    • 2017
  • BACKGROUND/OBJECTIVES: The anti-diabetic activity of pear through inhibition of ${\alpha}-glucosidase$ has been demonstrated. However, little has been reported about the effect of pear on insulin signaling pathway in obesity. The aims of this study are to establish pear pomace 50% ethanol extract (PPE)-induced improvement of insulin sensitivity and characterize its action mechanism in 3T3-L1 cells and high-fat diet (HFD)-fed C57BL/6 mice. MATERIALS/METHODS: Lipid accumulation, monocyte chemoattractant protein-1 (MCP-1) secretion and glucose uptake were measure in 3T3-L1 cells. Mice were fed HFD (60% kcal from fat) and orally ingested PPE once daily for 8 weeks and body weight, homeostasis model assessment of insulin resistance (HOMA-IR), and serum lipids were measured. The expression of proteins involved in insulin signaling pathway was evaluated by western blot assay in 3T3-L1 cells and adipose tissue of mice. RESULTS: In 3T3-L1 cells, without affecting cell viability and lipid accumulation, PPE inhibited MCP-1 secretion, improved glucose uptake, and increased protein expression of phosphorylated insulin receptor substrate 1 [p-IRS-1, ($Tyr^{632})$)], p-Akt, and glucose transporter type 4 (GLUT4). Additionally, in HFD-fed mice, PPE reduced body weight, HOMA-IR, and serum lipids including triglyceride and LDL-cholesterol. Furthermore, in adipose tissue, PPE up-regulated GLUT4 expression and expression ratio of p-IRS-1 ($Tyr^{632})/IRS$, whereas, down-regulated p-IRS-1 ($Ser^{307})/IRS$. CONCLUSIONS: Our results collectively show that PPE improves glucose uptake in 3T3-L1 cells and insulin sensitivity in mice fed a HFD through stimulation of the insulin signaling pathway. Furthermore, PPE-induced improvement of insulin sensitivity was not accompanied with lipid accumulation.

Effect of M11C (Non-lectin Components) Obtained from Korean Mistletoe on the $IL-1\beta$ Secretion from Mouse Splenocytes (쥐의 비장세포로부터 $IL-1\beta$ 분비에 있어서 한국산 겨우살이 추출물 M11C (비렉틴 구성물질)의 효과)

  • Jun, Myung-Ha;Kang, Tae-Bong;Chang, Sung-Ho;Choi, Wahn-Soo;Seong, Nak-Sul;Her, Erk
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.1
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    • pp.38-45
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    • 2007
  • Korean mistletoe (Viscum album L) extract has been found to posses immunoregulating activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract activates splenocytes to secret interleukin $1\beta(IL-1\beta)$. The splenocytes were treated with M11C, and then collected the supernatant and cell lysate that were prepared to analyze the level of $IL-1\beta$, using ELISA, immunoblotting, and RT-PCR. Maximum effective dose and time of M11C on $IL-1\beta$ secretion from splenocytes were $200{\mu}g/m\ell$ and 8 hours, respectively. Treatment dose and time for the maximum expression of $IL-1\beta$ mRNA were $200{\mu}g/m\ell$ and 4 hours, respectively. Saccharide degradation enzyme Viscozyme L completely blocked the effect of M11C on $IL-1\beta$ secretion from splenocytes. As the result, among non-lectin components saccharide could be regarded as a main component for $IL-1\beta$ expression from splenocytes.

Effects of Cinnamaldehyde on Salivary Gland Tissue in Xerostomia Model

  • Choi, Ja-Hyeong;Lee, Jung-Hwa;Kim, Yeon-Hwa;Hyun, Kyung-Yae;Park, Chung-Mu;Lee, Min-Kyung
    • Biomedical Science Letters
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    • v.26 no.2
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    • pp.93-100
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    • 2020
  • Xerostomia is a relatively common oral disease that causes various problems such as pain, discomforted, tissue damage, and infection. When the activity of AQPs, which plays an important role in the microbial channel transmembrane activity in tissues, decreases saliva secretion and the oral cavity dryness occurs. In this study, we observed whether there was a change in tissue through the expression level of AQP-5 in the submandibular gland in the 4-DAMP-induced xerostomia model. First, in order to construct a xerostomia model, 4-DAMP (1 mg/kg) and 20% urethane (0.5 mL/kg) were administered intraperitoneal (i.p.) to experimental animals. To observe the changes in the submandibular gland was excised, H&E staining was performed and protein quantitation analysis was performed using the submandibular tissue to observe the changes in AQP5 protein expression involved in changes in saliva secretion. Also, cinnamaldehyde (5, 12.5, 25 and 50 mg/kg) dissolved in 20% DMSO, in distilled water for each concentration, and then orally administered at a dose of 1 mL for biopsy and protein quantitative analysis. As a result, it was observed that the submandibular tissue, a model of xerostomia was wider than the naïve group. And then western blot analysis, the expression level of AQP5 decreased in the 4-DAMP group compared to the naïve group, and the expression increased in the group administered orally with cinnamaldehyde. Therefore, administration of 4-DAMP resulted in histological changes for xerostomia, and cinnamaldehyde would be a material that can be developed by reducing xerostomia.

Increase in Insulin Secretion Induced by Panax ginseng Berry Extracts Contributes to the Amelioration of Hyperglycemia in Streptozotocin-induced Diabetic Mice

  • Park, Eun-Young;Kim, Ha-Jung;Kim, Yong-Kyoung;Park, Sang-Un;Choi, Jae-Eul;Cha, Ji-Young;Jun, Hee-Sook
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.153-160
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    • 2012
  • Panax ginseng has long been used as a traditional herbal medicine. More recently, it has received attention for its anti-diabetic and anti-obesity effects in humans and in animal models of type 2 diabetes. In the present study, we tested the hypoglycemic effects of ginseng berry extract in beta-cell-deficient mice and investigated the mechanisms involved. Red (ripe) and green (unripe) berry extracts were prepared and administered orally (100 or 200 mg/kg body weight) to streptozotocin-induced diabetic mice daily for 10 wk. The body weight was measured daily, and the nonfasting blood glucose levels were measured after 5 and 10 wk after administration. Glucose tolerance tests were performed, and the serum insulin levels were measured. The proliferation of beta-cells was measured in vitro. The administration of red or green ginseng berry extract significantly reduced the blood glucose levels and improved the glucose tolerance in beta-cell deficient mice, with the higher doses resulting in better effects. Glucose-stimulated insulin secretion was significantly increased in berry extract-treated mice compared with streptozotocin-induced diabetic control mice. Treatment with ginseng berry extract increased beta-cell proliferation in vitro. Both red berry and green berry extracts improved glycemic control in streptozotocin-induced diabetic mice and increased insulin secretion, possibly due to increased beta-cell proliferation. These results suggest that ginseng berry extracts might have beneficial effects on beta-cell regeneration.

Common and differential effects of docosahexaenoic acid and eicosapentaenoic acid on helper T-cell responses and associated pathways

  • Lee, Jaeho;Choi, Yu Ri;Kim, Miso;Park, Jung Mi;Kang, Moonjong;Oh, Jaewon;Lee, Chan Joo;Park, Sungha;Kang, Seok-Min;Manabe, Ichiro;Ann, Soo-jin;Lee, Sang-Hak
    • BMB Reports
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    • v.54 no.5
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    • pp.278-283
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    • 2021
  • Our understanding of the differential effects between specific omega-3 fatty acids is incomplete. Here, we aimed to evaluate the effects of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) on T-helper type 1 (Th1) cell responses and identify the pathways associated with these responses. Naïve CD4+ T cells were co-cultured with bone marrow-derived dendritic cells (DCs) in the presence or absence of palmitate (PA), DHA, or EPA. DHA or EPA treatment lowered the number of differentiated IFN-γ-positive cells and inhibited the secretion of IFN-γ, whereas only DHA increased IL-2 and reduced TNF-α secretion. There was reduced expression of MHC II on DCs after DHA or EPA treatment. In the DC-independent model, DHA and EPA reduced Th1 cell differentiation and lowered the cell number. DHA and EPA markedly inhibited IFN-γ secretion, while only EPA reduced TNF-α secretion. Microarray analysis identified pathways involved in inflammation, immunity, metabolism, and cell proliferation. Moreover, DHA and EPA inhibited Th1 cells through the regulation of diverse pathways and genes, including Igf1 and Cpt1a. Our results showed that DHA and EPA had largely comparable inhibitory effects on Th1 cell differentiation. However, each of the fatty acids also had distinct effects on specific cytokine secretion, particularly according to the presence of DCs.

Expression of Human Lactoferrin Gene and Secretion in Saccharomyces diastaticus YIY345 (효모 Saccharomyces diastaticus YIY 345에서의 Human Lactoferrin 유전자 발현 및 분비)

  • Joo, Yun Jung;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.23 no.1
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    • pp.80-89
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    • 1996
  • The expression and secretion of human lactoferrin (hLf) in Sacclnromyces diastaticus were performed. 1. For the secretion of hLf in yeast, recombinant plasmid pYEGLf was constructed using promoter, secretion signal sequence of glucoamylase I gene (STA1) and transcriptional terminator of GAL7 gene. 2. Each correct recombinant plasmid was selected by mini-preparation of plasmid DNA from E coli transformant and restriction enzyme digestion analysis. The selected plasmids, pYEGLf, were transformed into S. diastaticus YIY345 as a expression host, respectively. 3. Western blot analysis using rabbit anti-hLf was carried out to identify expressed hLf. Positive signals were shown in culture supernatant of pYEGLf transformant. 4. About $100{\mu}g-1mg$ of concentrated culture supernatant of positive clone were loaded on paper disc and tested for the antimicrobial activity against E coli. However, no activity was observed. We concluded that this fact results from low concentration of hLf secreted from yeast, compared with the fact that MIC of hLf is as high as $3mg/m{\ell}$. Therefore, the purification of secreted hLf may be require to investigate the antimicrobial activity. From this study, the feasibility of low-cost production of sufficient quantities of human lactofferin for nutritional and therapeutical applications were suggested.

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Proteome Analysis of Bacillus subtilis When Overproducing Secretory Protein

  • Jang Mi;Park Byoung-Chul;Lee Do-Hee;Kho Chang-Won;Cho Sa-Yeon;Lee Baek-Rak;Park Sung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.368-373
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    • 2006
  • Bacillus subtilis and related Bacillus species are frequently used as hosts for the mass production of recombinant proteins. Accordingly, this study examined the cellular response of B. subtilis to the overexpression of a soluble secretory protein. As such, the lichenase derived from B. cereus was overexpressed in B. subtilis, initially localized in the cytoplasm as a mature form and then secreted into the medium. Thereafter, the proteome of B. subtilis was analyzed using 2D electrophoresis and MALDI-TOF mass spectrometry. The expression of several heat-shock proteins, such as dnaK and groEL, was increased under this condition. In addition, manganese superoxide dismutase and NADH dehydrogenase were also upregulated in the lichenase-secreting B. subtilis. Therefore, it was concluded that the transient accumulation of a secreted protein in B. subtilis before secretion acted as a stress on the cell, which in turn induced the expression of various protective proteins.

7α-Hydroxycholesterol Elicits TLR6-Mediated Expression of IL-23 in Monocytic Cells

  • Seo, Hyun Chul;Kim, Sun-Mi;Eo, Seong-Kug;Rhim, Byung-Yong;Kim, Koanhoi
    • Biomolecules & Therapeutics
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    • v.23 no.1
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    • pp.84-89
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    • 2015
  • We investigated the question of whether 7-oxygenated cholesterol derivatives could affect inflammatory and/or immune responses in atherosclerosis by examining their effects on expression of IL-23 in monocytic cells. $7{\alpha}$-Hydroxycholesterol ($7{\alpha}OHChol$) induced transcription of the TLR6 gene and elevated the level of cell surface TLR6 protein in THP-1 monocytic cells. Addition of an agonist of TLR6, FSL-1, to TLR6-expressing cells by treatment with $7{\alpha}OHChol$ resulted in enhanced production of IL-23 and transcription of genes encoding the IL-23 subunit ${\alpha}$ (p19) and the IL-12 subunit ${\beta}$ (p40). However, treatment with 7-ketocholesterol (7K) and $7{\beta}$-hydroxycholesterol ($7{\beta}OHChol$) did not affect TLR6 expression, and addition of FSL-1 to cells treated with either 7K or $7{\beta}OHChol$ did not influence transcription of the genes. Pharmacological inhibition of ERK, Akt, or PI3K resulted in attenuated transcription of TLR6 induced by $7{\alpha}OHChol$ as well as secretion of IL-23 enhanced by $7{\alpha}OHChol$ plus FSL-1. Inhibition of p38 MAPK or JNK resulted in attenuated secretion of IL-23. These results indicate that a certain type of 7-oxygenated cholesterol like $7{\alpha}OHChol$ can elicit TLR6-mediated expression of IL-23 by monocytic cells via PI3K/Akt and MAPKs pathways.

Efficacies of Potential Probiotic Candidates Isolated from Traditional Fermented Korean Foods in Stimulating Immunoglobulin A Secretion

  • Chang-Yong Choi;Chang-Hee Lee;Jun Yang;Seok-Jin Kang;In-Byung Park;Si-Won Park;Na-Young Lee;Hyun-Been Hwang;Hyun Sun Yun;Taehoon Chun
    • Food Science of Animal Resources
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    • v.43 no.2
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    • pp.346-358
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    • 2023
  • The aim of this study was to evaluate efficacies of selected lactic acid bacteria (LAB) in inducing immunoglobulin A (IgA) secretion. Twenty-five different LAB isolated from traditional fermented Korean foods were characterized for their probiotic properties and screened to identify those that could stimulate lamina propria cells (LPCs) from Peyer's patch to secret IgA in vitro. Among them, four strains (Lactiplantibacillus plantarum CJW55-10, Lactiplantibacillus pentosus CJW18-6, L. pentosus CJW56-11, and Pediococcus acidilactici CJN2696) were found to be strong IgA inducers. The number of IgA positive B cells and soluble IgA level were increased when LPCs were co-cultured with these LAB. Expression levels of toll-like receptor (TLR) such as TLR2 and TLR4 and secretion of interleuckin-6 were augmented in LPCs treated with these LAB. Further, we determined whether oral intake of these LAB enhanced IgA production in vivo. After one-week of daily oral administration, these LAB feed mice increased mucosal IgA and serum IgA. In conclusion, selected strains of LAB could induce systemic IgA secretion by activating lamina propria B cells in Peyer's patch and oral intake of selected strains of LAB can enhance systemic immunity by inducing mucosal IgA secretion.

Effects of Oral Administration of Phellinus linteus on the Productions of the Th1- and Th2-type Cytokines in Mice

  • Oh, Gi-Su;Pae, Hyun-Ock;Choi, Byung-Min;Kwon, Ji-Wung;Yun, Yeong-Ho;Choi, Jeong-Ho;Kwon, Tae-Oh;Park, Young-Chul;Chung, Hun-Teag
    • IMMUNE NETWORK
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    • v.3 no.3
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    • pp.182-187
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    • 2003
  • Background: The mushroom Phellinus linteus (PL) has been shown to have the anti-tumor and immunostimulatory effects. We hypothesized that the hot water extract of PL (WEPL) exerts its significant immunostimulatory effect by inducing production of the Th1-derived cytokine interferon-${\gamma}$ (IFN-${\gamma}$) by T lymphocytes. Methods: T lymphocytes were isolated from the mice fed with 200 mg/kg of WEPL once a day for 4 weeks, and then stimulated with the mitogen concanavaline A (Con A). IFN-${\gamma}$ gene and intracellular protein expressions were analyzed by RT-PCR and flow cytometry, respectively. The production of IFN-${\gamma}$ was measured by enzyme-linked immunosorbent assay. Results: WEPL significantly enhanced the transcription of IFN-${\gamma}$ mRNA. The effect of WEPL on IFN-${\gamma}$ expression was further supported by a concomitant increase in the number of cells with intracellular IFN-${\gamma}$ protein as well as the secretion of IFN-${\gamma}$. However, WEPL did not modulate either gene expression or protein secretion of interleukin-4, a Th2-associated cytokine, by Con A-stimulated T lymphocytes. Conclusion: Our results demonstrate that one of the potentially beneficial anti-tumor and immunostimulatory effects of WEPL may be mediated through the enhancement of IFN-${\gamma}$ secretion by T lymphocytes.