• 제목/요약/키워드: inducible proteins

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Characterization of a Low Molecular Weight Heat-Shock Protein cDNA Clone from Nicotiana tabacum

  • Park, Soo-Min;Joe, Myung-Kuk;Hong, Choo-Bong
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1999년도 춘계학술발표대회:발표논문요지록
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    • pp.18-18
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    • 1999
  • We characterized a cDNA clone for a low molecular weight heat-shock protein (LMW HSP) from tobacco named TLHS-l. Nucleotide sequence determination of TLHS-1 identified an open reading frame for 159 amino acids. To the upstream of the open reading frame, a sequence of 124 nucleotides was determined. To the 3' downstream of the open reading frame, 212 nucleotides were identified which carried poly(A)-tail. Comparison of the open reading frame and hydropathy plot of TLHS-1 with the previously reported class I LMW HSPs showed high identity which classified TLHS-1 as a class I LMW HSP cDNA clone. We proposed that there are six consensus regions in class I LMW HSPs. RNA blot hybridization for TLHS-1 showed a typical expression pattern of heat-shock-inducible gene from three common tobacco cultivars. The open reading frame of TLHS-1 was overexpressed in Escherichia coli. TLHS-1 protein confers thermal protection of other proteins in vitro and in vivo. Thermal induced aggregation of citrate synthase was reduced by purified TLHS-1 protein, and thermal death rate at $50^{\circ}C$ was reduced in E. coli expressing TLHS-l. From these data, we can expect that TLHS-1 acts as a molecular chaperone.perone.

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A Lipid-derived Endogenous Inducer of COX-2: a Bridge Between Inflammation and Oxidative Stress

  • Uchida, Koji
    • Molecules and Cells
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    • 제25권3호
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    • pp.347-351
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    • 2008
  • Several lines of evidence indicate that the oxidative modification of protein and the subsequent accumulation of the modified proteins have been found in cells during aging, oxidative stress, and in various pathological states including premature diseases, muscular dystrophy, rheumatoid arthritis, and atherosclerosis. The important agents that give rise to the modification of a protein may be represented by reactive aldehydic intermediates, such as ketoaldehydes, 2-alkenals and 4-hydroxy-2-alkenals. These reactive aldehydes are considered important mediators of cell damage due to their ability to covalently modify biomolecules, which can disrupt important cellular functions and can cause mutations. Furthermore, the adduction of aldehydes to apolipoprotein B in low-density lipoproteins (LDL) has been strongly implicated in the mechanism by which LDL is converted to an atherogenic form that is taken up by macrophages, leading to the formation of foam cells. During the search for an endogenous inducer of cyclooxygenase-2 (COX-2), an inducible isoform responsible for high levels of prostaglandin production during inflammation and immune responses, 4-hydroxy-2-noennal (HNE), one of the most representative lipid peroxidation product, has been identified as the potential inducer of COX-2. In addition, the following study on the molecular mechanism of the COX-2 induction by HNE has unequivocally established that a serum component, which is eventually identified to be denatured LDL, is essential for COX-2 induction. Here I review current understanding of the mechanisms by which HNE in cooperation with the serum component activates gene expression of COX-2.

RAW 264.7 대식세포 내에서 남정목 열매 추출물의 항염증 효과 (Anti-Inflammatory Effect of Extracts from Ligustrum obtusifolium S. fruits in RAW 264.7 Macrophages)

  • 문주호;고흥;신선미;김기태
    • 대한한의진단학회지
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    • 제17권3호
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    • pp.263-273
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    • 2013
  • Objectives This study was designed to investigate the anti-inflammatory effect of extracts from Ligustrum obtusifolium S. fruits(LOF) in RAW 264.7 Macrophages stimulated with lipopolysaccharide(LPS). Methods We examined productions of nitric oxide(NO), reactive oxygen species(ROS), inducible isoforms of NO synthase(iNOS), cyclooxygenase-2(COX-2) to investigate the anti-inflammatory effect of LOF extracts. In addition, we measured generation of pro-inflammatory cytokines(TNF-${\alpha}$, IL-6). Results Cell viability showed that LOF extracts had no cytotoxicity in Raw 264.7 cells. The treatment with LOF extracts significantly decreased the generation of NO and pro-inflammatory cytokines(TNF-${\alpha}$, IL-6) in LPS-stimulated macrophage cells. Furthermore LOF extracts inhibited intracellular ROS generation dose dependently and reduced the expression of iNOS, COX-2 proteins. Conclusions These results showed that the LOF extracts had an anti-inflammatory effect on LPS-stimulated Raw 264.7 cells. These findings provide scientific support for the use of this Ligustrum obtusifolium S. for inflammatory-related diseases.

Inhibitory Effect of Panax notoginseng and Berberine on LPS-induced iNOS, COX-2 and Prostaglandin E2

  • Lee, Boo-Young;Moon, Jin-Young;Kim, Cherl-Ho;Park, Won-Hwan
    • 동의생리병리학회지
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    • 제20권5호
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    • pp.1327-1333
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    • 2006
  • Many traditional herbal remedies exhibit several beneficial effects including anti-inflammation. Panax notoginseng Buck F.H. Chen. is used as a therapeutic agent to stop haemorrhages and a tonic to promote health in Korean and Chinese medicine. The pharrnacokinetic profiles of the main P. notoginseng are still not accurately investigated. The exact mechanism of the anti-inflammatory action of P. notoginseng, however, has not been determined. In the present study, we examined the effect of P. notoginseng on lipopolysaccharide (LPS)-induced nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene expression in RAW264.7 macrophages. The results indicated that P. notoginseng concentration-dependently inhibited LPS-induced NO production. Furthermore, P. notoginseng inhibited the expression of LPS-induced iNOS and COX-2 proteins without an appreciable cytotoxic effect on RAW264.7 cells. Berberine also inhibited LPS-induced iNOS protein as potently as P. notoginseng. This was consistent with the findings that P. notoginseng and also berberine inhibited prostaglandin E2 synthesis induced by LPS.

HIF-1-Dependent Induction of Jumonji Domain-Containing Protein (JMJD) 3 under Hypoxic Conditions

  • Lee, Ho-Youl;Choi, Kang;Oh, Hookeun;Park, Young-Kwon;Park, Hyunsung
    • Molecules and Cells
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    • 제37권1호
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    • pp.43-50
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    • 2014
  • Jumonji domain-containing proteins (JMJD) catalyze the oxidative demethylation of a methylated lysine residue of histones by using $O_2$, ${\alpha}$-ketoglutarate, vitamin C, and Fe(II). Several JMJDs are induced by hypoxic stress to compensate their presumed reduction in catalytic activity under hypoxia. In this study, we showed that an H3K27me3 specific histone demethylase, JMJD3 was induced by hypoxia-inducible factor (HIF)-$1{\alpha}/{\beta}$ under hypoxia and that treatment with Clioquinol, a HIF-$1{\alpha}$ activator, increased JMJD3 expression even under normoxia. Chromatin immunoprecipitation (ChIP) analyses showed that both HIF-$1{\alpha}$ and its dimerization partner HIF-$1{\beta}$/Arnt occupied the first intron region of the mouse JMJD3 gene, whereas the HIF-$1{\alpha}/{\beta}$ heterodimer bound to the upstream region of the human JMJD3, indicating that human and mouse JMJD3 have hypoxia-responsive regulatory regions in different locations. This study shows that both mouse and human JMJD3 are induced by HIF-1.

Construction of a Bile-responsive Expression System in Lactobacillus plantarum

  • Chae, Jong Pyo;Pajarillo, Edward Alain;Hwang, In-Chan;Kang, Dae-Kyung
    • 한국축산식품학회지
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    • 제39권1호
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    • pp.13-22
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    • 2019
  • This study aimed to develop a bile-responsive expression system for lactobacilli. The promoters of four genes, encoding phosphoenolpyruvate-dependent sugar phosphotransferase (mannose-specific), L-lactate dehydrogenase (LDH), HPr kinase, and D-alanine-D-alanine ligase, respectively, which were highly expressed by bile addition in Lactobacillus johnsonii PF01, were chosen. Each promoter was amplified by polymerase chain reaction and fused upstream of the ${\beta}$-glucuronidase gene as a reporter, respectively. Then, these constructs were cloned into E. coli-Lactobacillus shuttle vector pULP2, which was generated by the fusion of pUC19 with the L. plantarum plasmid pLP27. Finally, the constructed vectors were introduced into L. plantarum for a promoter activity assay. The LDH promoter showed the highest activity and its activity increased 1.8-fold by bile addition. The constructed vector maintained in L. plantarum until 80 generations without selection pressure. A bile-responsive expression vector, $pULP3-P_{LDH}$, for Lactobacillus spp. can be an effective tool for the bile-inducible expression of bioactive proteins in intestine after intake in the form of fermented dairy foods.

Nonstructural Protein of Severe Fever with Thrombocytopenia Syndrome Phlebovirus Inhibits TBK1 to Evade Interferon-Mediated Response

  • Lee, Jae Kyung;Shin, Ok Sarah
    • Journal of Microbiology and Biotechnology
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    • 제31권2호
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    • pp.226-232
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    • 2021
  • Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging phlebovirus of the Phenuiviridae family that has been circulating in the following Asian countries: Vietnam, Myanmar, Taiwan, China, Japan, and South Korea. Despite the increasing infection rates and relatively high mortality rate, there is limited information available regarding SFTSV pathogenesis. In addition, there are currently no vaccines or effective antiviral treatments available. Previous reports have shown that SFTSV suppresses the host immune response and its nonstructural proteins (NSs) function as an antagonist of type I interferon (IFN), whose induction is an essential part of the host defense system against viral infections. Given that SFTSV NSs suppress the innate immune response by inhibiting type I IFN, we investigated the mechanism utilized by SFTSV NSs to evade IFNmediated response. Our co-immunoprecipitation data suggest the interactions between NSs and retinoic acid inducible gene-I (RIG-I) or TANK binding kinase 1 (TBK1). Furthermore, confocal analysis indicates the ability of NSs to sequester RIG-I and related downstream molecules in the cytoplasmic structures called inclusion bodies (IBs). NSs are also capable of inhibiting TBK1-interferon regulatory factor 3 (IRF3) interaction, and therefore prevent the phosphorylation and nuclear translocation of IRF3 for the induction of type I IFN. The ability of SFTSV NSs to interact with and sequester TBK1 and IRF3 in IBs demonstrate an effective yet unique method utilized by SFTSV to evade and suppress host immunity.

Neuroprotective effects of paeoniflorin against neuronal oxidative stress and neuroinflammation induced by lipopolysaccharide in mice

  • Meng, Hwi Wen;Lee, Ah Young;Kim, Hyun Young;Cho, Eun Ju;Kim, Ji Hyun
    • Journal of Applied Biological Chemistry
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    • 제65권1호
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    • pp.23-31
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    • 2022
  • Oxidative stress and neuroinflammation play important roles in the pathogenesis of Alzheimer's disease (AD). This study investigated the protective effects of paeoniflorin (PF) against neuronal oxidative stress and neuroinflammation in lipopolysaccharide (LPS)-induced mice. The brains of LPS-injected control group showed significantly increased neuroinflammation by activating the nuclear factor kappa B (NF-κB) pathway and increasing inflammatory mediators. However, administration of PF significantly attenuated oxidative stress by inhibiting lipid peroxidation, nitric oxide levels, and reactive oxygen species production in the brain; PF at doses of 5 and 10 mg/kg/day downregulated the expression of NF-κB pathway-related proteins and significantly decreased inflammatory mediators including inducible nitric oxide synthase and cyclooxygenase-2. Moreover, the levels of brain-derived neurotrophic factor and its receptor, tropomycin receptor kinase B, were significantly increased in PF-treated mice. Furthermore, acetylcholinesterase activity and the ration of B-cell lymphoma 2 (Bcl-2)/Bcl-2 associated X were significantly reduced by PF in the brains of LPS-induced mice, resulting in the inhibition of cholinergic dysfunction and neuronal apoptosis. Thus, we can conclude that administration of PF to mice prevents the development of LPS-induced AD pathology through the inhibition of neuronal oxidative stress and neuroinflammation, suggesting that PF has a therapeutic potential for AD.

IFN-γ: A Crucial Player in the Fight Against HBV Infection?

  • Marine Laure Bettina Hillaire;Philip Lawrence;Brice Lagrange
    • IMMUNE NETWORK
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    • 제23권4호
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    • pp.30.1-30.18
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    • 2023
  • About 0.8 million people die because of hepatitis B virus (HBV) infection each year. In around 5% of infected adults, the immune system is ineffective in countering HBV infection, leading to chronic hepatitis B (CHB). CHB is associated with hepatocellular carcinoma, which can lead to patient death. Unfortunately, although current treatments against CHB allow control of HBV infection, they are unable to achieve complete eradication of the virus. Cytokines of the IFN family represent part of the innate immune system and are key players in virus elimination. IFN secretion induces the expression of interferon stimulated genes, producing proteins that have antiviral properties and that are essential to cell-autonomous immunity. IFN-α is commonly used as a therapeutic approach for CHB. In addition, IFN-γ has been identified as the main IFN family member responsible for HBV eradication during acute infection. In this review, we summarize the key evidence gained from cellular or animal models of HBV replication or infection concerning the potential anti-HBV roles of IFN-γ with a particular focus on some IFN-γ-inducible genes.

Anti-Inflammatory Effect of Freeze-Dried Broccoli Sprout Powder with Antioxidant Activity in RAW264.7 Cells

  • Hyun Jung Lim;Jong Soon Choi
    • 한국식생활문화학회지
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    • 제39권3호
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    • pp.156-165
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    • 2024
  • Sprout products, such as broccoli, alfalfa, and cabbage, have positive health effects. Thus far, sprout foods have attracted attention owing to their good bioavailability. In particular, young broccoli sprouts exhibit anti-inflammatory, antioxidant, and anti-cancer effects. They contain 100 times more chemoprotective substances than adult broccoli. This study examined the anti-inflammatory effects of freeze-dried young sprout broccoli (FD-YB) in vitro using RAW264.7 macrophage cells. The FD-YB powder antioxidant ability test showed that the radical-scavenging activity and superoxide dismutase enzyme activity increased in a dose-dependent manner. In addition, FD-YB was not cytotoxic to RAW264.7 cells, and nitric oxide production decreased after the FD-YB treatment of lipopolysaccharide-stimulated RAW264.7 cells in a dose-dependent manner. Furthermore, FD-YB significantly decreased the expression of inflammation-related proteins (Cyclooxygenase-2, Inducible nitric oxide synthase, and Prostaglandin E Synthase 2) and cytokines (Tumor necrosis factor-α and Interleukin-6). In conclusion, FD-YB can be a potential nutraceutical for preventing and regulating excessive immune responses during inflammation.