• Title/Summary/Keyword: ${\alpha}$-oxidase

Search Result 199, Processing Time 0.03 seconds

Role of Reactive Oxygen Species in Transforming Growth Factor-β1-inuduced Fibronectin Secretion and α-Smooth Muscle Actin Expression in Human Lung Fibroblasts (사람 폐 섬유아세포의 전환성장인자-β1에 의한 fibronectin 분비와 α-smooth muscle actin 표현에 있어서 활성산소족의 역할)

  • Ha, Hunjoo;Yu, Mi-Ra;Uh, Soo-taek;Park, Choon Sik;Lee, Hi Bahl
    • Tuberculosis and Respiratory Diseases
    • /
    • v.58 no.3
    • /
    • pp.267-276
    • /
    • 2005
  • Background : The transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$) plays a key role in lung fibrosis. However, the molecular mechanisms involved in $TGF-{\beta}1$-induced lung fibrosis are unclear. $TGF-{\beta}1$ is the key inducer of myofibroblast transdifferentiation via de novo synthesis of ${\alpha}-smooth$ muscle actin (${\alpha}-SMA$). Since $TGF-{\beta}1$ signals through reactive oxygen species (ROS) and ROS have been shown to induce accumulation of extracellular matrix (ECM) in various tissues, this study examined if ROS play a role in $TGF-{\beta}1$-induced fibronectin secretion and ${\alpha}-SMA$ expression in human lung fibroblasts, MRC-5 cells. Methods : Growth arrested and synchronized MRC-5 cells were stimulated with $TGF-{\beta}1$ (0.2-10 ng/ml) in the presence or absence of N-acetylcysteine (NAC) or diphenyleneiodonium (DPI) for up to 96 hours. Dichlorofluorescein (DCF)-sensitive cellular ROS were measured by FACScan and secreted fibronectin and cellular ${\alpha}-SMA$ by Western blot analysis. Results : $TGF-{\beta}1$ increased the level of fibronectin secretion and ${\alpha}-SMA$ expression in MRC-5 cells in a dosedependent manner. Both NAC (20 and 30 mM) and DPI (1 and $5{\mu}M$) significantly inhibited $TGF-{\beta}1$-induced fibronectin and ${\alpha}-SMA$ upregulation. The $TGF-{\beta}1$-induced cellular ROS level was also significantly reduced by NAC and DPI. Conclusions : The results suggest that NADPH oxidase-dependent ROS play an important role in $TGF-{\beta}1$-induced fibronectin secretion and ${\alpha}-SMA$ expression in MRC-5 cells, which leads to myofibroblast transdifferentiation and progressive lung fibrosis.

Purification and Characterization of Cytochrome c Oxidase from Photosynthetic Bacterium, Rhodopseudomonas gelatinosa (광합성세균 Rhodopseudomonas gelatinosa 의 시토크롬 c 산화효소의 정제 및 특성)

  • 강대길;최원기
    • Korean Journal of Microbiology
    • /
    • v.30 no.2
    • /
    • pp.101-107
    • /
    • 1992
  • Cytochrome c oxida5e from chemotrophically grown R p , geliitinosu was purified by cytochrome c affinity chromatography and DEAE-Sephacel ion exchange chromatography. The molecular weight of the cytochrome c oxidase was approximately 110.000 Da by sephacryl s-300 gel chromatography and approximately 52, 000 Da by SDS-gel electrophoresis, respectively. Therefore. cytochrolne c oxidase of Rps. gehtinosu seems to be dimer. The cytochrome c oxidasc was very sensitive to temperature. It's Km and Vmax were 20 pM and 44 unitlmg protein for horsc heart cytochrome c as a substrate. respectively, and its optimum pH and temperature were 6.4 and 25$^{\circ}$C. respectively. The absorption peaks of the reduced cytochrome c oxidase showed at 554 nm, 523 nm. and 422 nm. The activiiy of cytochrome c oxidase was inhibited by KCN, and NaN3, but not by CO, antimycir~ A. and myxothiazol. The cytochrome c-551 was produced either in phototrophically or chemotrophically grown Rps. gelaiinosci. The rcduced cytochrome c-551 was oxidized by b-type cytochrome c oxidase from Rp.v. gc.lrtino.sc~. Km and Vmax of cytochrome c oxidase was 26 pM and 31 unitlnlg protein For cytochrome c-551 as a substrate. respectively. Thercfore. thc electron transfer chain of chemotrophically grown Rps. glatinosa seems lo be ubiquinol cytochrome bc, complex -'cytochrome c-55lMb-type cytochrome c oxidase+02.

  • PDF

Effect of Occlusion on the Activities of Dermal Xanthine Oxidase in Rats (흰쥐에 있어서 피부노출 차단이 피부조직의 Xanthine Oxidase 활성에 미치는 영향)

  • 한선일;전태원;윤종국
    • Biomedical Science Letters
    • /
    • v.6 no.1
    • /
    • pp.37-43
    • /
    • 2000
  • To investigate an impact of skin occlusion on the dermal xanthine oxidase (XO) activity, the dorsal part in rats was covered with closed petri dish-shaped chamber, 46 mm in diameter and 10 mm in height, which was made of glass. The crack between top of chamber and skin was sealed by an adhesive agent. After 5 days, the quantity of sweat accumulation was about 400 mg, whereas after 10 days that was decreased about to 21 mg. The 5 days skin occlusion showed the more increased activity of dermal XO compared with the control, and the increased ratio of enzyme activity to the control was higher than that of 10 days skin occlusion, with the increase being associated with sweat accumulation in chamber. Furthermore, the V$_{max}$ of dermal XO in 5 days skin occlusion was higher than that in the control. In conclusion, it may be hypothesized that the XO system may play an role for defence mechanism in dermal tissue.

  • PDF

Liver PPAR${\alpha}$ and UCP2 are Involved in the Regulation of Ovariectomy-Induced Adiposity and Steatosis by Swim Training

  • Jeong, Sun-Hyo;Yoon, Mi-Chung
    • Biomedical Science Letters
    • /
    • v.16 no.4
    • /
    • pp.239-246
    • /
    • 2010
  • It is suggested that ovariectomy induces body weight gain primarily in the form of adipose tissue in rodents. Since liver peroxisome proliferator-activated receptor ${\alpha}$ (PPAR${\alpha}$) and uncoupling 2 (UCP2) are involved in the regulation of energy expenditure, it was investigated whether swim training regulates ovariectomy-induced adiposity and steatosis through liver PPAR${\alpha}$ and UCP2 activation in female ovariectomized mice, an animal model of postmenopausal women. Swim-trained mice had significantly decreased adipose tissue weights compared with sedentary control mice. Histological analysis showed that hepatic lipid accumulation was inhibited by swim training. Concomitantly, swim training significantly increased mRNA levels of PPAR${\alpha}$ and its target genes responsible for peroxisomal fatty acid ${\beta}$-oxidation, such as acyl-CoA oxidase, enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase and thiolase in the liver. Moreover, swim training induced the mRNA expression of UCP2. These results suggest that swim training can effectively prevent adiposity and steatosis caused by ovariectomy, in part through activation of liver PPAR${\alpha}$ and UCP2 in female obese mice.

Physicochemical Characteristics and Antioxidant Activity of Bracken (Pteridium aquilinum Kuhn) in Namhae (남해산 고사리의 이화학적 특성 및 항산화 활성)

  • Sim, Hye-Jin;Hwang, Cho-Rong;Kang, Jae-Ran;Kang, Min-Jung;Shin, Jung-Hye
    • Korean journal of food and cookery science
    • /
    • v.31 no.3
    • /
    • pp.288-295
    • /
    • 2015
  • In this study, we investigate the physicochemical characteristics of dried bracken (Pteridium aquilinum Kuhn) harvested in Namhae. The moisture, ash, crude protein and crude lipid content was $10.79{\pm}0.31%$, $6.16{\pm}0.04%$, $33.20{\pm}0.40%$ and $2.45{\pm}0.27%$, respectively. The total mineral (Ca, Fe, K, Mg, Na, Mn) content was $36720.1{\pm}495.7mg/kg$, with K being the highest at $31890.0{\pm}503.8mg/kg$. The total free amino acid content was 704.41 mg/100 g, the amount of methionine, citrulline and sarcosine being higher than the others. The water and 50% ethanol extracts from the dried bracken were evaluated for the total poly phenolic compound content, antioxidant activity and xanthine oxidase and ${\alpha}$-glucosidase inhibitory activity. The total polyphenol content in the 50% ethanol extract ($1574.86{\pm}18.31mg/100g$) was higher than in the water extract ($1240.24{\pm}16.32mg/100g$). The DPPH and ABT radical scavenging activity was increased according to the concentration of bracken extract, and the activity in the 50% ethanol extract was higher than in the water extract. In addition, the xanthine oxidase and ${\alpha}$-glucosidase inhibitory activity was also higher in the 50% ethanol extract than in the water extract. In conclusion, bracken has great antioxidative and antidiabetic effects and can be used as a preventive agent for oxidation and diabetes.

Antihyperlipidemic Effect of Ginsenoside Rg1 in Type 2 Diabetic Mice (제2형 당뇨병 모델 마우스에서 ginsenoside Rg1의 항당뇨 효과)

  • Park, Jae-Hong;Lee, Ji-Youn;Yeo, Ji-Young;Nam, Jeong-Su;Jung, Myeong-Ho
    • Journal of Life Science
    • /
    • v.21 no.7
    • /
    • pp.932-938
    • /
    • 2011
  • Ginsenoside Rg1 is a pharmacologically active component isolated from ginseng. The goal of this study was to clarify the beneficial effects of Rg1 on glucose and lipid metabolism in diabetic animals (db/db mice). To accomplish this, ten week old db/db mice were administered 10 mg/kg of Rg1 for 15 days. Rg1 did not influence the weight of db/db mice when compared with vehicle-treated db/db mice. The administration of Rg1 lowered fasting plasma glucose, and improved glucose tolerance. Importantly, Rg1 markedly reduced both plasma triglyceride and free fatty acids, and increased high-density lipoprotein cholesterol (HDL-C) concentrations in db/db mice. Rg1 activated promoter activity of chimeric GAL4-PPAR${\alpha}$ reporter and increased expression of peroxisome proliferator-activated receptor alpha (PPAR${\alpha}$) target genes such as carnitine palmitoyltransferase-1 (CPT-1) and acyl-CoA oxidase (ACO), which are involved in fatty acid oxidation. These findings indicated that improvement of lipid profiles by Rg1 may be associated with increased fatty acid oxidation via PPAR${\alpha}$ activation. Taken together, these results suggest that Rg1 could have beneficial effects for controlling hyperglycemia and hyperlipidemia associated with type 2 diabetes.

Effects of $\alpha$-Tocopherol and Perilla oil on the Toxicity of Polychlorinated biphenyl in Rat (랏트에 대한 Polychlorinated Biphenyl의 독성에 미치는 $\alpha$-Tocopherol과 Perilla oil의 효과)

  • 최경현;김문석;황두환;문재규;김성오
    • Environmental Analysis Health and Toxicology
    • /
    • v.3 no.3_4
    • /
    • pp.39-51
    • /
    • 1988
  • Effects of ${\alpha}$-tocopherol and perilla oil on the toxicity of polychlorinated biphenyls (PCB) in male rat were studied. Rats were fed ad libitum for 6 weeks with the animal diet which contains PCB 30 ppm and 100 ppm. Perilla oil (0.5 g/kg body weight) and ${\alpha}$-tocopherol (30 mg/kg body weight) were administered intraperitoneally twice a week for 6 weeks. Rats fed with PCB showed enlargement of liver and spleen, increase in aspartate aminotransferase, alkaline phosphatase, sereum lipid and cytochrome P 450 and decrease in body weight and glutathione. When perilla oil was administered to rats fed with PCB increase in aspartate aminotransferase, alkaline phosphatase, serum lipid and cytochrome P45O and decrease in body weight and glutathione were significantly augmented, compared to rats fed with PCB alone. This means that perilla oil potentiates the toxicity of PCB. On the other hand when ${\alpha}$-tocopherol was administered to rats fed with PCB increase in aspartate aminotransferase, alkaline phosphatase, serum lipid and cytochrome P45O and decrease in body weight and glutathione were signigicantly reduced, compared to rats fed with PCB alone. This means that u-tocopherol reduces the toxicity of PCB. From the above results, it may be concluded that PCB is metabolized by microsomal mixed function oxidase and the metabolite causes the toxicity and microsomal glutathione plays a role of protection on the toxicity of PCB.

  • PDF

Studies on the ROS(Reactive Oxygen Species)-Scavenging Activities of DDMP saponins Isolated from Glycine max (L.) Merrill (대두 (Glycine max (L.) merrill.)에서 추출한 DDMP 사포닌의 활성산소종 제거 작용)

  • 조수진;백희준;이상선;정일민;하지희;강주섭;고현철;신인철;이창호
    • Biomolecules & Therapeutics
    • /
    • v.8 no.1
    • /
    • pp.32-37
    • /
    • 2000
  • Recently new soybean saponins with D DMP (2,5-dihydroxy-6-methyl-2,3,- dihydro-4H-pyran-4-one) moiety have been isolated from legumes. The purpose of this study is to characterize ROS scavenging activities of DDMP saponins ($\alpha$g, $\beta$g saponin) isolated from Glycine max (L.) Merrill. The scavenging activity on OH was examined in terms of lipid peroxidation in the rat liver homogenates and the same activity on $O_2$ was also determined in the xanthine-xanthine oxidase system, respectively. Up to 0.25 mg DDMP saponins ($\alpha$g and $\beta$g saponins) did not cause any significant effects on the prevention of lipid peroxidation as compared with the control group. In terms of superoxide scavenging activities, 0.25 and 0.5 mg $\alpha$g saponin inhibits only 2.6% and 5.5% (p<0.05) of the control group, respectively. However, $\alpha$g saponin dose-dependently (p<0.01, r=0.955) inhibits the formation of superoxide radical unto 21.3% of the control group with a maximal dose of 0.5 mg (p<0.01), equivalent to 0.17 units of superoxide dismutase activity.

  • PDF

Isolation of Polyphenol from Green Tea by HPLC and Its Physiological Activities (HPLC에 의한 녹차의 polyphenol 화합물의 분리 및 polyphenol의 생리활성)

  • Woo, Hee-Seob;Choi, Hee-Jin;Han, Ho-Suk;Park, Jung-Hye;Son, Jun-Ho;An, Bong-Jeun;Son, Gyu-Mok;Choi, Cheong
    • Korean Journal of Food Science and Technology
    • /
    • v.35 no.6
    • /
    • pp.1199-1203
    • /
    • 2003
  • Polyphenols were isolated from Korean green tea using Sephadex LH-20 and HPLC. The isolated polyphenols were procyanidin B-4, procyanidin B-2-3,3'-digallate, prodelphinidin C-2-3,3'-di-O-digallate, (+)-catechin-3-O-rhamnose, procyandin B-5, procyanidin B-7-3-0-gallate, gallate, epiafzelechin-$(4{\beta}{\rightarrow}8)$-epiafzelechin, procyanidin B-3-3-O-rhamnose, afzelechin-$(4{\alpha}{\rightarrow}8)$-catechin, prodelphinidin B-5-3,3'-di-O-digallate and (+)-taxifolin-3-O-D-xyloside. The inhibitory effects of prodelphinidin C-2-3,3'-di-O-gallate and procyanidin B-2-3,3'-digallate $(at\;100{\mu}M)$ on angiotensin.converting enzyme were 68.8 and 54.6%, respectively, while the inhibitory effects of prodelphinidin C-2-3,3'-di-O-gallated and procyanidin B-2-3,3'-digallate $(at\;100{\mu}m)$ on xanthine oxidase were 54.5 and 38.2%, respectively. Lastly, the inhibitory activities of prodelphinidin C-2-3,3'-di-O-gallate $(at\;100{\mu}m)$ on tyrosinase was 42.1%.

High-level Secretory Expression of Recombinant $\beta$-Agarase from Zobellia galactanivorans in Pichia pastoris (Pichia pastoris에서 Zobellia galactanivorans 유래 재조합 $\beta$-Agarase의 고효율 분비생산)

  • Seok, Ji-Hwan;Park, Hee-Gyun;Lee, Sang-Hyeon;Nam, Soo-Wan;Jeon, Sung-Jong;Kim, Jong-Hyun;Kim, Yeon-Hee
    • Microbiology and Biotechnology Letters
    • /
    • v.38 no.1
    • /
    • pp.40-45
    • /
    • 2010
  • The gene encoding $\beta$-agarase (agaB) which hydrolyzes $\beta$-1,4 linkages of agarose from Zobellia galactanivorans was cloned and fused to Saccharomyces cerevisiae mating factor alpha-1 secretion signal ($MF{\alpha}1$), in which the transcription of $MF{\alpha}1$-AgaB was under the control of AOX1 (alcohol oxidase 1, methanol inducible) promoter. The constructed plasmid pPIC-AgaB (9 kb) was integrated into HIS4 gene locus of Pichia pastoris genome. Successful integration was confirmed by performing colony PCR. The transformed cells showed red halos around its colonies in methanol agar plate by adding iodine solution, indicating the active expression of agaB in P.pastoris. By SDS-PAGE and zymographic analysis, the molecular weight of $\beta$-agarase was estimated to be a 53 kDa and about 15% N-linked glycosylation was occurred. The activity of extracellular $\beta$-agarase reached 1.34, 1.42 and 1.53 units/mL by inducing 0.1, 0.5, and 1% methanol, respectively, at baffled flask culture of P.pastoris GS115/pPIC-AgaB for 48 hr. Most of the enzyme activity was found in the extacellular fraction and the secretion efficiency showed 98%. Thermostability of recombinant $\beta$-agarase was also increased by glycosylation.