• Title/Summary/Keyword: gene regulation

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Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

Growth performance, carcass traits, muscle fiber characteristics and skeletal muscle mRNA abundance in hair lambs supplemented with ferulic acid

  • Pena-Torres, Edgar Fernando;Castillo-Salas, Candelario;Jimenez-Estrada, Ismael;Muhlia-Almazan, Adriana;Pena-Ramos, Etna Aida;Pinelli-Saavedra, Araceli;Avendano-Reyes, Leonel;Hinojosa-Rodriguez, Cindy;Valenzuela-Melendres, Martin;Macias-Cruz, Ulises;Gonzalez-Rios, Humberto
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.52-69
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    • 2022
  • Ferulic acid (FA) is a phytochemical with various bioactive properties. It has recently been proposed that due to its phytogenic action it can be used as an alternative growth promoter additive to synthetic compounds. The objective of the present study was to evaluate the growth performance, carcass traits, fiber characterization and skeletal muscle gene expression on hair-lambs supplemented with two doses of FA. Thirty-two male lambs (n = 8 per treatment) were individually housed during a 32 d feeding trial to evaluate the effect of FA (300 and 600 mg d-1) or zilpaterol hydrochloride (ZH; 6 mg d-1) on growth performance, and then slaughtered to evaluate the effects on carcass traits, and muscle fibers morphometry from Longissimus thoracis (LT) and mRNA abundance of β2-adrenergic receptor (β2-AR), MHC-I, MHC-IIX and IGF-I genes. FA increased final weight and average daily gain with respect to non-supplemented animals (p < 0.05). The ZH supplementation increased LT muscle area, with respect to FA doses and control (p < 0.05). Cross-sectional area (CSA) of oxidative fibers was larger with FA doses and ZH (p < 0.05). Feeding ZH increased mRNA abundance for β2-AR compared to FA and control (p < 0.05), and expression of MHC-I was affected by FA doses and ZH (p < 0.05). Overall, FA supplementation of male hair lambs enhanced productive variables due to skeletal muscle hypertrophy caused by MHC-I up-regulation. Results suggest that FA has the potential like a growth promoter in lambs.

In vitro Anti-oxidative and Anti-inflammatory Activities of Horse-bone Extract via Up-regulation of Heme-oxygenase 1 (말뼈추출물의 Hemeoxygenase-1의 발현 조절을 통한 시험관내 항염증 효과)

  • Im, Eun Ju;Lee, Ki-Ja;Cho, Gil-Jae;Kim, Hyun-Kyoung;Kim, Suk;Rhee, Man Hee
    • Journal of agriculture & life science
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    • v.50 no.2
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    • pp.139-150
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    • 2016
  • Few studies have been reported that horse-bone extract(HBE) can prevent and treatment of bone diseases. However, HBE' therapeutic activities are still not fully understood. This study determined whether HBE up-regulates hemeoxygenase 1(HO-1) and this mediates its anti-inflammatory effect in murine macrophages.Nitric oxide(NO) assay, MTT assay and DPPH assay were performed. In addition, Western blotting and real time PCR were used to determine protein expression, and gene expression, respectively. HBE significantly inhibited NO production without observable cytotoxicity. In addition, HBE attenuated inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2) and phospho (p)-ERK protein expressions in LPS(0.1㎍/ml) stimulated RAW264.7 cells. On the other hand, HBE alone up-regulated HO-1 and Nrf-2 expressions, which mediated HBE's anti-inflammatory effect in RAW264.7 cells. Finally, HBE up-regulated HO-1 and impaired ERK1/2 signaling pathways, and thus it may provide protection against cellular oxidation and inflammation.

Metabolic Regulation of Longevity and Immune Response in Caenorhabditis elegans by Ingestion of Lacticaseibacillus rhamnosus IDCC 3201 Using Multi-Omics Analysis

  • Daniel Junpyo Lee;Ju Young Eor;Min-Jin Kwak;Junbeom Lee;An Na Kang;Daye Mun;Hyejin Choi;Minho Song;Jong Nam Kim;Jun-Mo Kim;Jungwoo Yang;Hyung Wook Kim;Sangnam Oh;Younghoon Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1109-1118
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    • 2024
  • Probiotics, specifically Lacticaseibacillus rhamnosus, have garnered attention for their potential health benefits. This study focuses on evaluating the probiotic properties of candidate probiotics L. rhamnosus IDCC 3201 (3201) using the Caenorhabditis elegans surrogate animal model, a well-established in vivo system for studying host-bacteria interactions. The adhesive ability to the host's gastrointestinal tract is a crucial criterion for selecting potential probiotic bacteria. Our findings demonstrated that 3201 exhibits significantly higher adhesive capabilities compared with Escherichia coli OP50 (OP50), a standard laboratory food source for C. elegans and is comparable with the widely recognized probiotic L. rhamnosus GG (LGG). In lifespan assay, 3201 significantly increased the longevity of C. elegans compared with OP50. In addition, preconditioning with 3201 enhanced C. elegans immune response against four different foodborne pathogenic bacteria. To uncover the molecular basis of these effects, transcriptome analysis elucidated that 3201 modulates specific gene expression related to the innate immune response in C. elegans. C-type lectin-related genes and lysozyme-related genes, crucial components of the immune system, showed significant upregulation after feeding 3201 compared with OP50. These results suggested that preconditioning with 3201 may enhance the immune response against pathogens. Metabolome analysis revealed increased levels of fumaric acid and succinic acid, metabolites of the citric acid cycle, in C. elegans fed with 3201 compared with OP50. Furthermore, there was an increase in the levels of lactic acid, a well-known antimicrobial compound. This rise in lactic acid levels may have contributed to the robust defense mechanisms against pathogens. In conclusion, this study demonstrated the probiotic properties of the candidate probiotic L. rhamnosus IDCC 3201 by using multi-omics analysis.

A Novel Human BTB-kelch Protein KLHL31, Strongly Expressed in Muscle and Heart, Inhibits Transcriptional Activities of TRE and SRE

  • Yu, Weishi;Li, Yongqing;Zhou, Xijin;Deng, Yun;Wang, Zequn;Yuan, Wuzhou;Li, Dali;Zhu, Chuanbing;Zhao, Xueying;Mo, Xiaoyang;Huang, Wen;Luo, Na;Yan, Yan;Ocorr, Karen;Bodmer, Rolf;Wang, Yuequn;Wu, Xiushan
    • Molecules and Cells
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    • v.26 no.5
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    • pp.443-453
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    • 2008
  • The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.

Up-regulation of Pluripotency-related Genes in Human Amniotic Fluid-derived Stem Cells by ESRRB Conjugated with Cell-Penetrating Peptide (인간 양수 유래 줄기세포에서 세포투과단백질을 이용한 ESRRB 단백질의 직접도입에 의한 전분화능 관련 유전자의 발현 조절)

  • Jo, Jung-Hyun;Lee, Yu-Sun;Oh, Mi-Hee;Ko, Jung-Jae;Cheon, Yong-Pil;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.14 no.4
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    • pp.243-251
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    • 2010
  • ESRRB (Estrogen related receptor $\beta$) is an orphan receptor, and have a role on maintaining the undifferentiated state and self-renewal of pluripotent stem cell as a transcription factor which regulates the expression of OCT4 and NANOG genes. Also, Feng et al. (2009) reported that Esrrb, Oct4 and Sox2 could induce pluripotent stem cell from somatic cells. The aim of the present study was to develop the direct delivery system of human ESRRB protein into human amniotic fluid-derived stem cells (AFSCs) and to analyze the effect of ESRRB on the regulation of pluripotency-related genes. Human ESRRB has three isoforms arisen by alternative splicing. We cloned short-form ESRRB and made a fusion protein of ESRRB and R7 for an efficient protein transfer to cell. R7 as cell-penetrating peptide(CPP) can help to transfer ESRRB into cells. R7-ESRRB-His6 protein was observed in the cytoplasm and nuclei within 5 hours after treatment. Also, we could observe R7-ESRRB-His6 protein only in the nuclei within 24 hours. Realtime PCR showed that ESRRB increased expression of OCT4 and NANOG as well as SOX2 gene. Therefore, we demonstrated that R7-ESRRB-His6 proteins were efficiently transferred into the nuclei of AFSCs and work well as a possible transcription factor.

Development of Lotus Root Bugak with Plasma Lipid Reduction Capacity by Addition of Opuntia ficus-indica var. saboten or Green Tea as a Coloring Agent (백년초 및 녹차 가루 첨가 연근 부각의 지질저하 기능성)

  • Kim, Mijeong;Hong, Sun Hee;Chung, Lana;Choe, Eunok;Song, Yeong-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.333-340
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    • 2014
  • The purpose of this study was to develop functional lotus root bugak with plasma lipid reduction capacity by controlling the color of batter used for bugak preparation. Lotus root, nearly colorless, was selected to observe color effects. Gardeniae fructus (GF), Opuntia ficus-indica var. saboten (OF), and green tea (GT), which are colored yellow, red, and green, respectively, were used as coloring agents. Fermented glutinous rice was prepared naturally during winter season by placing glutinous rice and water (1:2, w/w) together in a crock pot for 7 days. Coloring materials (10%, w/w) were blended with glue made from fermented glutinous rice flour to prepare the batter. Cooked lotus root was then mixed with a 1.1-fold amount of batter (w/w) and dried at room temperature. Lotus root bugak (LRB) is pan-fried with un-roasted sesame oil, which is traditionally used as frying oil in Korea. Low-density lipoprotein receptor knockout ($LDLr^{-/-}$) mice (n=36) were fed an atherogenic diet (AD) containing various types of LRB (10 g%) for 10 weeks. Plasma triglyceride, total cholesterol, and LDL-C concentrations decreased significantly in mice fed LRB prepared with OF batter (OFB) and GT batter (GTB) (P<0.05). Protein expression levels of fatty acid synthase (FAS) and 3-hydroxyl-3-methylglutaryl coenzyme A reductase (HMGCR) in the OFB and GTB groups were suppressed compared with the LRB group (P<0.05). In accordance with the results on FAS and HMGCR expression, sterol regulatory element binding protein-I and II (SREBP-I and II), which are responsible for the regulation of FAS and HMGCR gene expression, respectively, were down-regulated compared to the LRB group (P<0.05). In conclusion, the plasma lipid reduction activities of OFB and GTB could be mediated through down-regulation of FAS and HMGCR mRNA expression via suppression of regulatory molecules, SREBP-I and II, in $LDLr^{-/-}$ mice.

Expression of Bombyx mori Transferrin Gene in Response to Oxidative Stress or Microbes (미생물 및 산화적 스트레스에 의한 누에 트랜스페린 발현)

  • Yun, Eun-Young;Kwon, O-Yu;Hwang, Jae-Sam;Ahn, Mi-Young;Goo, Tae-Won
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1607-1611
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    • 2011
  • To analyze the role of Bombyx mori transferrin (BmTf) in response to microbes or oxidative stress, we investigated the level of BmTf transcripts in B. mori treated with various microbes and oxidative stress inducers. BmTf mRNA was mainly expressed in the epidermis and fat in the bodies of B. mori injected with Escherichia coli, and up regulated in response to microbes such as bacteria, fungi, or viruses, but was hardly altered in response to oxidative stress inducers such as $H_2O_2$, Cu, or $FeCl_3$. We also confirmed that BmTf mRNA expression was increased in Bm5 cells treated with ERK, PLC, PKA, PI3K, MAPK, or JNK inhibitors, respectively. To identify the major inducer of BmTf expression, we analyzed the amount of serum iron in the hemolymph of B. mori after injection or feeding with E. coli or $FeCl_3$. The results showed that the amount of serum iron was not changed by injection and feeding with E. coli, although BmTf mRNA was increased by injection with E. coli. On the contrary, injection and feeding with $FeCl_3$ significantly increased the amount of serum iron, although they did not alter the BmTf mRNA level. On the basis of these results, we assume that up-regulation of BmTf in B. mori is closely related to the defense of microorganism, and BmTf may be expressed at the basal constitutive level when it plays a role in iron metabolism by maintaining iron homeostasis and in the insect defense mechanism against oxidative stress.

Anti-inflammatory Effects of Rebaudioside A in LPS-stimulated RAW264.7 Macrophage Cells (LPS에 의해 자극된 RAW264.7 대식세포에서 Rebaudioside A의 항염 효과)

  • Cho, Uk Min;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.157-164
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    • 2017
  • Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.

A Study on the Gene Expression in Shikonin-Induced Inhibition of Adipogenesis (Shikonin에 의한 지방세포형성 억제과정에서의 유전자 발현 연구)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Chung, Sang-In;Cho, Soo-Hyun;Oh, Dong-Jin;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1637-1643
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    • 2009
  • Shikonin, a component of Lithospermum erythrorhizon Sieb. et Zucc, exerts various characteristics such as anti-inflammatory, anti-cancer and anti-obesity functions. To elucidate the molecular mechanism of shikonin-induced inhibition of adipogenesis, we analyzed the mRNA expression level of various adipogenesis-related factors including C/EBPs (CCAAT/enhancerbinding proteins) and $PPAR{\gamma}$ (peroxisome proliferator-activated receptor $\gamma$). The data showed that mRNA expressions of C/$EBP{\beta}$ and C/$EPB{\delta}$ were only slightly changed by shikonin treatment, but mRNA expressions of $PPAR{\gamma}$ and C/$EPB{\alpha}$ were significantly down-regulated. Then, we tested whether upstream regulators of C/$EBP{\beta}$ and $PPAR{\gamma}$ were involved in anti-adipogenesis of shikonin. C/$EBP{\gamma}$ and CHOP (C/EBP homologous protein), which are upstream regulators of C/$EBP{\beta}$, were not affected by shikonin treatment. On the contrary, the mRNA level of KROX20 was markedly down-regulated by shikonin treatment. These results suggest that KROX20 might regulate downstream factors of adipogenesis through C/$EBP{\beta}$-independent pathway. The expression of KLF15 (Kruppel-like factor15), which is a member of KLF family and is a upstream regulator of $PPAR{\gamma}$, was dramatically decreased by shikonin treatment, but KLF2 was not changed. Shikonin had no impact on the expression of KLF5 in the early stage of adipogenesis, but shikonin increased expression of KLF5 in the late stage of adipogenesis. Even though mRNA expression of KLF5 was moderately changed by shikonin treatment, its effect may be small compared with the effect of KLF15, which was markedly inhibited. Taken together, these results suggest that shikonin inhibits adipogenesis through the down-regulation of $PPAR{\gamma}$ and C/$EPB{\alpha}$, which is mediated by the down-regulation of two pro-adipogenic factors, KROX20 and KLF15.