The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.
Oxidative stress has been implicated in cutaneous damage in various inflammatory skin diseases, including atopic dermatitis. The present study was undertaken to investigate the antioxidative and anti-inflammatory activities of the extract of Duchesnea chrysantha (DCE). DEC was prepared by extracting with 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Total flavonoid and polyphenol contents of DCE were included $24.73{\pm}0.45$ and $178.77{\pm}2.65$, respectively. DCE significantly increased electron donating ability (DPPH), nitrite scavenging (NO) and ABTS reducing activity in dose dependant. We investigated the anti-inflammatory effects of DCE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. DCE significantly suppressed NO and prdstaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with DCE in a dose dependent manner. These results suggest that DEC may has value as natural product with its high quality functional components, antioxidative and anti-inflammatory activities.
Objectives: To investigate the effects of Chrysanthemum indicum L. pharmacopuncture on lipids, antioxidant capacity and anti-inflammation in rats fed high-fat diet. Methods: Hyperlipidemic rats induced by high-fat diet were divided into 5 groups: no treatment control (normal, n=8), high-fat diet only control (control, n=8), high-fat diet and Chrysanthemum indicum L. pharmacopuncture at CV4 group (TI, n=8), high-fat diet and Chrysanthemum indicum L. pharmacopuncture at CV17 group (TII, n=8), and high-fat diet and Chrysanthemum indicum L. pharmacopuncture at EX-HN3 group (TIII, n=8). They were given pharmacopuncture accordingly every other day for two weeks followed by analyses of lowering lipids effects, oxidative capacity and anti-inflammatory effects. Results: Compared with the control, pharmacopuncture groups showed significantly decreased plasma total cholesterol (TC), liver thiobarbituric acid reactive substance (TBARS), catalase, glutathione peroxidase, neutrophils, monocytes, plasma and liver IL-$1{\beta}$, and plasma and liver IL-6. In other parameters including plasma and liver triglyceride, liver TC, LDL-cholesterol, HDL-cholesterol, liver TBARS, supraoxide dismutase, total protein, albumin, blood cell analysis, plasma and liver TNF-$\alpha$, and IL-10, there was no significant difference between control and pharmacopuncture groups. No clear acupoint-specificity was observed. Conclusions: Chrysanthemum indicum L. pharmacopuncture may improve control of hyperlipidemia.
This study was conducted to investigate the functional composition and antioxidant activity of Cirsium and Carduus genera based on different parts. Leaves of Cirsium setidens Nakai contained 23.66% protein and seeds of Carduus crispus L contained 25.30% lipid. Extraction yields of Cirsium and Carduus genera were higher in leaves than in any other parts of the plants. Total phenolics and total flavonoid content were abundant in extracts of leaves, steam and root of C. japonicum var. ussuriense, and the flower extract of C. setidens Nakai. Silymarin was not found in extracts of Cirsium and Carduus genera. Acacetin was identified in leaf or flower extracts of C. setidens Nakai, or in leaf and steam extracts of C. pendulum Fisch ex DC. Apigenin was identified in the flower extracts of Cirsium and Carduus genera and constituted 7.16 mg/g in C. japonicum var. ussuriense. Cynarin was present at 5.55 mg/g in the seed extract of C. setidens Nakai, and narirutin represented 19.56 mg/g and, 4.18 mg/g of the seed extracts of C. pendulum Fisch ex DC and, Carduus crispus L, respectively. 2,2-Diphenyl-1-2-picrylhydrazyl (DPPH) radical scavenger activity was higher in flower extracts of C. setidens Nakai than in the other genera. Photochemiluminescence activity was 2.3 nmol in leaves and flower extracts of C. setidens Nakai and flower extract of Carduus crispus L. Flowers and seeds of C. setidens Nakai shoe potential as new functional materials.
This study investigated the nutritional compositions and physiochemical properties of three domestically cultivated jujube fruits (Boeun, Sangwhang, and Bokjo). Nutrient contents were analyzed by freeze-drying the jujube fruits. The results showed that the proximate compositions of the three jujube cultivars ranged from 7.85~13.52% for moisture, 4.65~5.24% for crude protein, 1.29~2.38% for crude ash, 0.17~1.09% for crude lipid, and 7.66~16.63% for crude fiber. The above proximate contents of jujube cultivars were significantly different. Mineral contents of Al, Fe, Mg, Ca, Na, and K in the three jujube fruits were also determined. Mn, Cu and Zn were not detected in any of the varieties. The content of Al was higher in Sangwhang, while Boeun had higher contents of Ca and K. The total polyphenol, ABTS values, and hypoglycemic effect were slightly higher in Boeun as compared to the other varieties. Taken together, Boeun could be an important source of natural antioxidant as a functional food.
This study was conducted to provide insight into the management and care of the elderly in nursing homes. Eighty-six elderly subjects over 65 years old (38 male, 48 female) in 3 non-fee-paying nursing homes, located in Daejeon and Chungchong areas, were studied. Subjects'dietary intakes by estimated food intake records, and self-perceived health conditions, were studied using questionnaires from July 21 to August 1, 1997. Twenty-one % of the male subjects and 42% of the female subjects were over 80 years. Their predominant past occupation was farming. While 8.3% of the female and no male residents showed a BMI (Body Mass Index) of 27 and above, about 30% of the subjects were underweight and in poor health status in seeing, joint pains, lumbago and shoulder pains. Regarding overall health status, 72.1% of the subjects considered them to be in poor health, and female and male subjects suffered more difficulties from cloudy eyes, joint pains and lumbago and shoulder pains than any other. Neuralgia was the predominant chronic complaint and followed by hypertension in both sexes. Overall, female subjects felt worse off than the male subjects in terms of their health status, that can be attributed to higher average age of the female subjects compared to the male subjects. The elderly were eating a very low fat (about 15 g : 6% of total calorie) diet with low vitamin A and E. Intakes of calories, protein and iron slightly exceeded RDA. The phosphorus intake was more than double the RDA although calcium intake was close to the RDA. From these results, it seems important to doubling the fat intake of the elderly residents in the form of vegetable fat with the object of raising of vitamin E, antioxidant vitamin, and essential fatty acids for the elderly. It is also recommended that the elderly residents should be given adequate calcium and exercise for bone health.
Objective : This study aimed to evaluate the protective effect of Artemisiae Capillaris Herba (AC) in reflux esophagitis (RE) rats. Methods : The AC was measured antioxidant activity through in vitro experiments, such as total polyphenol and flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Base on the results, we had conducted in vivo experiments. Rats were divided normal, control, AC treatment 50 mg/kg BW (AC50), and AC treatment 100 mg/kg BW (AC100) groups. AC were orally administered 2 h before the induction of RE. RE was induced by tie the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats. The rats were sacrificed 5 h after the surgery. We analyzed the expression of inflammatory related markers by western blot and observed the production of reactive oxygen species (ROS) and hematoxylin-eosin staining, Results : The $IC_{50}$ of AC for DPPH and ABTS were showed 12.60 and $33.32{\mu}g/m{\ell}$ respectively. In the RE rat, AC decreased inflammatory related markers, such as phosphorylated inhibitor of ${\kappa}B{\alpha}$, nuclear factor-kappa B, cyclooxygenase-2, inducible nitric oxide synthase, and tumor necrosis factor alpha. Also, AC reduced the increased reactive oxygen species in serum. The anti-inflammatory effect of AC appeared to be partially mediated through the inhibition of ROS. Also, AC markedly ameliorated esophageal mucosa damage via the inhibition of protein expression related to inflammation. Conclusions : Therefore, these results suggest that AC would be used as a therapeutic material in protection and/or treatment for reflux esophagitis.
Park, Mi-Young;Kim, Min Young;Seo, Young Rok;Kim, Jong-Sang;Sung, Mi-Kyung
Journal of Cancer Prevention
/
제21권2호
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pp.95-103
/
2016
Background: Excess energy supply induces chronic low-grade inflammation in association with oxidative stress in various tissues including intestinal epithelium. The objective of this study was to investigate the effect of high-fat diet (HFD) on intestinal cell membrane integrity and intestinal tumorigenesis in $Apc^{Min/+}$ mice. Methods: Mice were fed with either normal diet (ND) or HFD for 12 weeks. The number of intestinal tumors were counted and biomarkers of endotoxemia, oxidative stress, and inflammation were determined. Changes in intestinal integrity was measured by fluorescein isothiocyanate (FITC)-dextran penetration and membrane gap junction protein expression. Results: HFD group had significantly higher number of tumors compared to ND group (P < 0.05). Blood total antioxidant capacity was lower in HFD group, while colonic 8-hydroxy-2'-deoxyguanosine level, a marker of oxidative damage, was higher in HFD group compared to that of ND group (P < 0.05). The penetration of FITC-dextran was substantially increased in HFD group (P < 0.05) while the expressions of membrane gap junction proteins including zonula occludens-1, claudin-1, and occludin were lower in HFD group (P < 0.05) compared to those in ND group. Serum concentration of lipopolysaccharide (LPS) receptor (CD14) and colonic toll-like receptor 4 (a LPS receptor) mRNA expression were significantly higher in HFD group than in ND group (P < 0.05), suggesting that significant endotoxemia may occur in HFD group due to the increased membrane permeability. Serum interleukin-6 concentration and myeloperoxidase activity were also higher in HFD group compared to those of ND group (P < 0.05). Conclusions: HFD increases oxidative stress disrupting intestinal gap junction proteins, thereby accelerating membrane permeability endotoxemia, inflammation, and intestinal tumorigenesis.
Zhen, Ao Xuan;Piao, Mei Jing;Kang, Kyoung Ah;Fernando, Pincha Devage Sameera Madushan;Kang, Hee Kyoung;Koh, Young Sang;Yi, Joo Mi;Hyun, Jin Won
Biomolecules & Therapeutics
/
제27권6호
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pp.562-569
/
2019
Niacinamide (NIA) is a water-soluble vitamin that is widely used in the treatment of skin diseases. Moreover, NIA displays antioxidant effects and helps repair damaged DNA. Recent studies showed that particulate matter 2.5 ($PM_{2.5}$) induced reactive oxygen species (ROS), causing disruption of DNA, lipids, and protein, mitochondrial depolarization, and apoptosis of skin keratinocytes. Here, we investigated the protective effects of NIA on $PM_{2.5}$-induced oxidative stress in human HaCaT keratinocytes. We found that NIA could inhibit the ROS generation induced by $PM_{2.5}$, as well block the $PM_{2.5}$-induced oxidation of molecules, such as lipids, proteins, and DNA. Furthermore, NIA alleviated $PM_{2.5}$-induced accumulation of cellular $Ca^{2+}$, which caused cell membrane depolarization and apoptosis, and reduced the number of apoptotic cells. Collectively, the findings show that NIA can protect keratinocytes from $PM_{2.5}$-induced oxidative stress and cell damage.
The protective adaptive response to electrophiles and reactive oxygen species is mediated by the induction of phase II detoxifying genes including glutathione S-transferases (GSTs). NF-E2-related factor-2 (Nrf2) phosphorylation by protein kinase C (PKC) is a critical event for its nuclear translocation in response to oxidative stress. Previously, we have shown that peroxynitrite plays a role in activation of Nrf2 and Nrf2 binding to the antioxidant response element (ARE) via the pathway of phosphatidylinositol 3-kinase (PI3-kinase) and that nitric oxide synthase in hepatocytes is required for GSTA2 induction. In view of the importance of PKC and Pl3-kinase in Nrf2-mediated GST induction, we investigated the role of these kinases in peroxynitrite formation for GSTA2 induction by oxidative stress and determined the relationship between PKC and PI3-kinase. Although PKC activation by phorbol 12-myristate-13-acetate (PMA) did not increase the extents of constitutive and inducible GSTA2 expression, either PKC depletion by PMA or PKC inhibition by staurosporine significantly inhibited GSTA2 induction by tert-butylhydroquinone (t-SHa) a prooxidant chemical. Therefore, the basal PKC activity is req- uisite for GSTA2 induction. 3-Morpholinosydnonimine (SIN-1), which decomposes and yields peroxynitrite, induced GSTA2, which was not inhibited by PKC depletion, but slightly enhanced by PKC activation, suggesting that PKC promotes peroxynitrite formation for Nrf2-mediated GSTA2 induction. Treatment of cells with S-nitroso-N-acetyl-penicillamine (SNAP), an exogenous NO donor, in combination with t-BHQ may produce peroxynitrite. GSTA2 induction by SNAP + t-BHQ was not decreased by PKC depletion, but rather enhanced by PKC activation, showing that the activity of PKC might be required for peroxynitrite formation. LY294002 a P13-kinase inhibitor blocked GSTA2 induction by t-BHQ, which was reversed by PMA-induced PKC activation. These results provide evidence that PKC may playa role in formation of peroxynitrite that activates Nrf2 for GSTA2 induction and that PKC may serve an activator for GSTA2 induction downstream of PI3-kinase.
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