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Fraxetin Induces Heme Oxygenase-1 Expression by Activation of Akt/Nrf2 or AMP-activated Protein Kinase α/Nrf2 Pathway in HaCaT Cells

  • Kundu, Juthika;Chae, In Gyeong;Chun, Kyung-Soo
    • Journal of Cancer Prevention
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    • v.21 no.3
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    • pp.135-143
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    • 2016
  • Background: Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, has been reported to possess antioxidative, anti-inflammatory and neuroprotective effects. A number of recent observations suggest that the induction of heme oxygenase-1 (HO-1) inhibits inflammation and tumorigenesis. In the present study, we determined the effect of fraxetin on HO-1 expression in HaCaT human keratinocytes and investigated its underlying molecular mechanisms. Methods: Reverse transcriptase-PCR and Western blot analysis were performed to detect HO-1 mRNA and protein expression, respectively. Cell viability was measured by the MTS test. The induction of intracellular reactive oxygen species (ROS) by fraxetin was evaluated by 2′,7′-dichlorofluorescin diacetate staining. Results: Fraxetin upregulated mRNA and protein expression of HO-1. Incubation with fraxetin induced the localization of nuclear factor-erythroid-2-related factor-2 (Nrf2) in the nucleus and increased the antioxidant response element-reporter gene activity. Fraxetin also induced the phosphorylation of Akt and AMP-activated protein kinase $(AMPK){\alpha}$ and diminished the expression of phosphatase and tensin homolog, a negative regulator of Akt. Pharmacological inhibition of Akt and $AMPK{\alpha}$ abrogated fraxetin-induced expression of HO-1 and nuclear localization of Nrf2. Furthermore, fraxetin generated ROS in a concentration-dependent manner. Conclusions: Fraxetin induces HO-1 expression through activation of Akt/Nrf2 or $AMPK{\alpha}/Nrf2$ pathway in HaCaT cells.

Production of Inflammatory Cytokines and Nitric Oxide by Human Mast Cells Incubated with Toxoplasma gondii Lysate

  • Park, Eun-Ah;Han, Ik-Hwan;Kim, Jung-Hyun;Park, Soon-Jung;Ryu, Jae-Sook;Ahn, Myoung-Hee
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.201-206
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    • 2019
  • The roles of mast cells in allergic diseases and helminth infections are well known. However, the roles of mast cells in T. gondii infection is poorly understood. This study was focused on the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-4), chemokines (CXCL8, MCP-1) and nitric oxide (NO) by mast cells in response to soluble lysate of T. gondii tachyzoites. Production of CXCL8 (IL-8), MCP-1, $TNF-{\alpha}$ and IL-4 were measured by RT-PCR and ELISA. Western blot were used for detection of CXCR-1 and CXCR2. Our results showed that T. gondii lysates triggered mast cells to release CXCL8, MCP-1, $TNF-{\alpha}$, IL-4 and to produce NO. This suggests that mast cells play an important role in inflammatory responses to T. gondii.

Changes in Microflora, Enzyme Activities and Microscopic Structure of Waxy Rice and Steeping Water in Response to Different Steeping Conditions During Preparation of Gangjung (강정 제조시 찹쌀의 수침조건에 따른 찹쌀과 수침액의 미생물상, 효소활성 및 미세구조 변화)

  • Kim, Haeng-Ran;Kim, Kyung-Mi;Kim, Kwang-Ok
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.644-651
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    • 2009
  • This study was conducted to investigate the effects of steeping periods and temperatures of waxy rice on the microflora and enzyme activities of steeping water and waxy rice, as well as on the microscopic structures of waxy rice during the preparation of gangjung. When the steeping water was 15 and $35^{\circ}C$, yeast and lactic acid bacteria were the major microflora after 1 day. After 11 days, Lactobacillus spp. were the major bacteria, while Bacillus spp. were the primary microflora after 21 days. At $25^{\circ}C$ the primary species were Bacillus spp., Leuconostoc spp., and Bacillus spp. after 1, 11 and 21 days, respectively. The $\alpha$-amylase activity of steeping water increased proportionally with the increase of temperature and period, while the protease activity tended to increase for 11 days, but decreased at 21 days. The enzyme activities of the steeped waxy rice, especially the $\alpha$-amylase activity, decreased with higher steeping temperature, while the $\beta$-amylase and glucoamylase activity remained constant for up to 21 days. Observation using scanning electron microscopy indicated that intracellular cleavage of steeped waxy rice endosperm resulted in a rough surface morphology due to the exposure of polygonal starch granules. Tiny pinholes on the surface of waxy rice starch granules were also observed after steeping for 11 days.

Effects of steamed Polygonatum odoratum extract on inhibition of adipocyte differentiation and lowing lipid in 3T3-L1 adipocytes (증자 둥굴레 추출물의 3T3-L1 지방세포에서 분화억제 및 지질강하 효과)

  • Kang, Byung Tae;Choe, Won Kyung;Park, Dong Cheol;Kim, Jong Kuk;Park, Mora;Kim, Sung Ok;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.29 no.2
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    • pp.15-21
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    • 2014
  • Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.

Tumor Necrosis Factor-Alpha $(TNF-{\alpha})$ Induces PTEN Expression in HL-60 Cells (백혈병세포에서 종양괴사인자에 의한 PTEN 발현증가)

  • Lee Seung-Ho;Park Chul-Hong;Kim Byeong-Su
    • Journal of Food Hygiene and Safety
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    • v.21 no.3
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    • pp.181-188
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    • 2006
  • Tumor necrosis factor-alpha $(TNF-{\alpha})$ plays a variety of biological functions such as apoptosis, inflammation and immunity. PTEN also has various cellular function including cell growth, proliferation, migration and differentiation. Thus, possible relationships between two molecules are suggested. $(TNF-{\alpha})$has been known to downregulate PTEN via nuclear factor-kappa $B(NF-{\kappa}B)$ pathway in the human colon cell line, HT-29. However, here we show the opposite finding that $(TNF-{\alpha})$ upregulates PTEN via activation of $NF-{\kappa}B$ in HL-60 cells. $TNF-{\alpha}$ increased PTEN expression at HL-60 cells in a time- and dose-dependent manner, but the response was abolished by disruption of $NF-{\kappa}B$ with p65 anisense oligonucleotide or pyrrolidine dithiocarbamate (PDTC). We found that $TNF-{\alpha}$ activated the $NF-{\kappa}B$ pathways, evidenced by the translocation of p65 to the nucleus in $TNF-{\alpha}-treated$ cells. We conclude that $TNF-{\alpha}$ induces upregulation of PTEN expression through $NF-{\kappa}B$ activation in HL-60 cells.

The Metabolic Effects of FGF21: From Physiology to Pharmacology (생리, 약학적 관점에서 fibroblast growth factor 21 (FGF21)의 대사 효과 고찰)

  • Song, Parkyong
    • Journal of Life Science
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    • v.30 no.7
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    • pp.640-650
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    • 2020
  • Fibroblast growth factor 21 (FGF21) is an atypical member of the FGF protein family which is highly synthesized in the liver, pancreas, and adipose tissue. Depending on the expression tissue, FGF21 uses endo- or paracrine features to regulate several metabolic pathways including glucose metabolism and energy homeostasis. Different physiologically stressful conditions such as starvation, a ketogenic diet, extreme cold, and mitochondrial dysfunction are known to induce FGF21 synthesis in various tissues to exert either adaptive or defensive mechanisms. More specifically, peroxisome proliferator-activated receptor gamma and peroxisome proliferator-activated receptor alpha control FGF21 expression in adipose tissue and liver, respectively. In addition, the pharmacologic administration of FGF21 has been reported to decrease the body weight and improve the insulin sensitivity and lipoprotein profiles of obese mice and type 2 diabetes patients meaning that FGF21 has attracted huge interest as a therapeutic agent for type 2 diabetes, obesity, and non-alcoholic fatty liver disease. However, understanding FGF21 remains complicated due to the paradoxical condition of its tissue-dependent expression. For example, nutrient deprivation largely increases hepatic FGF21 levels whereas adipose tissue-derived FGF21 is increased under feeding condition. This review discusses the issues of interest that have arisen from existing publications, including the tissue-specific function of FGF21 and its action mechanism. We also summarize the current stage of a clinical trial using several FGF21 analogs.

Study on relationship between "cholestrol·cardiorenal connective tissue weakness type" and TNF-alpha gene polymorphism in iris constitution analysis (홍채체질 분석에서 콜레스테롤·심신이 약한 체질과 TNF-alpha 유전자 다형성과의 상관성 연구)

  • Yoo, Chun-sang;Hwang, Woo-jun;Kim, Kyung-sik;Choi, Sung-yong;Kim, Jong-uk
    • Journal of Acupuncture Research
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    • v.21 no.2
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    • pp.89-101
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    • 2004
  • 홍채학은 대체의학의 한 분야로서 홍채 침착의 불규칙성을 주시함으로써 의학적 상태를 진단한다. 홍채학적 분류에 의한 체질은 가족력을 보이고 있으며 이는 홍채체질의 유전성을 의미한다. 강력한 면역조절자이며 전 염증성 사이토카인인 종양괴사인자(tumor necrosis factor-a, TNF-${\alpha}$)는 많은 병리적 과정에서 중요한 역할을 한다. 따라서 본 연구자는 고혈압환자에서 홍채체질과 TNF-${\alpha}$ 유전자 다형성과의 관련성을 조사하였다. 87명의 고혈압 환자와 79명의 정상인을 홍채체질에 따라 분류하였으며 이들의 TNF-${\alpha}$ 유전자형을 분석하였다. 결과적으로 정상인에 비하여 TNF-${\alpha}$ GA 이형접합체의 빈도가 고혈압 환자군에서 감소하였다. 이 같은 결과는 TNF-${\alpha}$ 다형성이 고혈압에 대한 저항성과 관련 있음을 의미한다. 또한 고혈압환자에서 콜레스테롤 침착체질과 심신 결합조직 허약 체질은 42.5%로서 정상인 16.5%에 비하여 현저하게 증가하였다 (P<0.001). GG TNF-${\alpha}$ 유전자형을 갖고 있는 군에서 심신 결합조직 허약체질과 콜레스테롤 침착체질의 빈도는 정상인보다 환자에서 유의하게 높았다(P<0.001). 본 연구에서 저자는 홍채체질과 고혈압사이의 관련성을 발견함과 동시에 TNF-${\alpha}$ 유전자 다형성과 고혈압, 그리고 홍채 체질과의 관련성을 최초로 입증하였다.

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Study on Tumor Necrosis Factor- ${\alpha}$ Gene Polymorphism in Rheumatoid Arthritis (류마티스 관절염에 있어 종양괴사인자 다형성에 대한 연구)

  • Kim, Kyung-Un;Han, Mi-Yung;Lee, Yoon-Kyung;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Journal of Acupuncture Research
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    • v.24 no.3
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    • pp.197-205
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    • 2007
  • Objectives : Tumor necrosis factor-${\alpha}$(TNF ${\alpha}$) is a proinflammatory cytokine involved in the pathogenesis of rheumatoid arthritis. This study was designed to investigate the relation between TNF-${\alpha}$ gene polymorphism and rheumatoid arthritis in Korean population. Methods : This study was carried out on 103 rheumatoid arthritis patients who fulfilled the American College of Rheumatology 1987 revised criteria for rheumatoid arthritis and 208 healthy control subjects. Blood samples from all subjects were obtained for DNA extraction. The extracted DNA was amplified by polymerse chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. We investigated the genotyping of TNF-${\alpha}$ by using Pyrosequencing. Results: The genotypes of TNF-${\alpha}$ gene were GG, AG and AA. While the distribution of TNF-${\alpha}$ polymorphism in control subjects was 92.31%, 7.21%, 0.48% respectively, in rheumatoid arthritis patients was 93.20%, 6.80%, 0.00%(GG, AG, AA). There was no statistical significant allelic frequency difference between control and rheumatoid arthritis groups. Conclusions : We concluded that there was no significant association between TNF-${\alpha}$ gene polymorphism and rheumatoid arthritis. However, the findings of this study need to be confirmed in more patients and further studies.

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Polymorphism Identification, RH Mapping and Association of ${\alpha}$-Lactalbumin Gene with Milk Performance Traits in Chinese Holstein

  • Zhang, Jian;Sun, Dongxiao;Womack, J.E.;Zhang, Yi;Wang, Yachun;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1327-1333
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    • 2007
  • Lactose synthase catalyses the formation of lactose which is the major osmole of bovine milk and regulates the milk volume. Alpha-lactalbumin (${\alpha}$-LA) is involved in the synthesis of lactose synthase in the mammary gland. Therefore ${\alpha}$-LA is regarded as a plausible candidate gene for the milk yield trait. To determine whether ${\alpha}$-LA is associated with milk performance traits, 1,028 Chinese Holstein cows were used to detect polymorphisms in the ${\alpha}$-LA by means of single-strand conformation polymorphism (SSCP). Two nucleotide transitions were identified in the 5'flanking region and intron 3 of ${\alpha}$-LA. Associations of such polymorphisms with five milk performance traits were analyzed using a general linear model procedure. No significant associations were observed between these polymorphisms and the five milk performance traits (p>0.05). RH mapping placed ${\alpha}$-LA on BTA5q21, linked most closely to markers U63110, CC537786 and L10347 (LOD>8.3), which is far distant from the region of the quantitative trait locus (QTL) on bovine chromosome 5 for variation in the milk yield trait. In summary, based on our findings, we eliminated these SNPs from having an effect on milk performance traits.

Inhibition of Over-oxidation of 11$\beta$-l7$\alpha$, 21-trihydroxy-pregna-1, 4-diene-3, 2-dione in Fermentative Process. (Prednisolone 발효중의 산화분해 저지법)

  • Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.2 no.3
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    • pp.127-131
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    • 1974
  • Inhibition of over-oxidation of prednisolone in the fermentation has been studied by using vegetative cells as enzyme source. firstly, A. simplex (ATCC 6946) was demonst.ated to degrade p.ednisolone in the vegetative culture of the microorganism. Over 72% of hydrocortisone was transformed into prednisoloneby 3 hours of the fermentation. However, the prednisolone produced was considerably oxidized forming over-oxidation product in 8hours of fermentation period with the intact cells. Secondly, in order to depress the over-oxidation and the breakdown of the steroid skeleton of prednisolone, chelating agents such as $\alpha$$\alpha$'-dipyridyl, o-phenanthroline and 8-hydroxyruinoline were added to the fermentation broth. Conseauently, the breakdown of prednisolone by the iutact cells was able to be remarkably retatded and an intermediate regarded as an oxidized product derived from prednisolone was accumulated, by the addition of $\alpha$$\alpha$'-dipytidyl in the fermentation.

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