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      Ecklonia cava Polyphenol Has a Protective Effect against Ethanol-Induced Liver Injury in a Cyclic AMP-Dependent Manner

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          Abstract

          Previously, we showed that Ecklonia cava polyphenol (ECP) treatment suppressed ethanol-induced increases in hepatocyte death by scavenging intracellular reactive oxygen species (ROS) and maintaining intracellular glutathione levels. Here, we examined the effects of ECP on the activities of alcohol-metabolizing enzymes and their regulating mechanisms in ethanol-treated hepatocytes. Isolated hepatocytes were incubated with or without 100 mM ethanol. ECP was dissolved in dimethylsulfoxide. ECP was added to cultured cells that had been incubated with or without ethanol. The cells were incubated for 0–24 h. In cultured hepatocytes, the ECP treatment with ethanol inhibited cytochrome P450 2E1 (CYP2E1) expression and activity, which is related to the production of ROS when large quantities of ethanol are oxidized. On the other hand, ECP treatment with ethanol increased the activity of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase. These changes in activities of CYP2E1 and ADH were suppressed by treatment with H89, an inhibitor of protein kinase A. ECP treatment with ethanol enhanced cyclic AMP concentrations compared with those of control cells. ECP may be a candidate for preventing ethanol-induced liver injury via regulating alcohol metabolic enzymes in a cyclic AMP-dependent manner.

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          Most cited references38

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          Antioxidant activities of enzymatic extracts from brown seaweeds.

          Potential antioxidative activities of enzymatic extracts from seven species of brown seaweeds were evaluated using four different reactive oxygen species (ROS) scavenging assays containing DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical, superoxide anion, hydroxyl radical and hydrogen peroxide scavenging assay. The brown seaweeds were enzymatically hydrolyzed to prepare water-soluble extracts by using five carbohydrate degrading enzymes (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) of commercial and inexpensive enzymes obtained from Novozyme Co. (Novozyme Nordisk, Bagsvaerd, Denmark). The enzymatic extracts exhibited more prominent effects in hydrogen peroxide scavenging activity (approximately 90%) compared to the other scavenging activities and the activity of enzymatic extracts was even higher than that of the commercial antioxidants. In particular, Ultraflo and Alcalase extracts of S. horneri were dose-dependent and thermally stable. Moreover the two enzymatic extracts strongly inhibited DNA damage (approximately 50%). Those extracts showed significantly (p<0.05) remarkable scavenging effects in DPPH free radical scavenging assay and the activity indicated a marked correlation with phenolic contents. From the results, enzymatic extracts of the brown seaweeds might be valuable antioxidative sources.
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            Alcohol, oxidative stress and free radical damage.

            The involvement of free radical mechanisms in the pathogenesis of alcoholic liver disease (ALD) is demonstrated by the detection of lipid peroxidation markers in the liver and the serum of patients with alcoholism, as well as by experiments in alcohol-feed rodents that show a relationship between alcohol-induced oxidative stress and the development of liver pathology. Ethanol-induced oxidative stress is the result of the combined impairment of antioxidant defences and the production of reactive oxygen species by the mitochondrial electron transport chain, the alcohol-inducible cytochrome P450 (CYP) 2E1 and activated phagocytes. Furthermore, hydroxyethyl free radicals (HER) are also generated during ethanol metabolism by CYP2E1. The mechanisms by which oxidative stress contributes to alcohol toxicity are still not completely understood. The available evidence indicates that, by favouring mitochondrial permeability transition, oxidative stress promotes hepatocyte necrosis and/or apoptosis and is implicated in the alcohol-induced sensitization of hepatocytes to the pro-apoptotic action of TNF-alpha. Moreover, oxidative mechanisms can contribute to liver fibrosis, by triggering the release of pro-fibrotic cytokines and collagen gene expression in hepatic stellate cells. Finally, the reactions of HER and lipid peroxidation products with hepatic proteins stimulate both humoral and cellular immune reactions and favour the breaking of self-tolerance during ALD. Thus, immune responses might represent the mechanism by which alcohol-induced oxidative stress contributes to the perpetuation of chronic hepatic inflammation. Together these observations provide a rationale for the possible clinical application of antioxidants in the therapy for ALD.
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              Bactericidal activity of phlorotannins from the brown alga Ecklonia kurome.

              The bactericidal activity of phlorotannins from brown algae against food-borne pathogenic bacteria (25 strains), methicillin-resistant Staphylococcus aureus (MRSA) (nine strains) and Streptococcus pyogenes (one strain) was examined and compared with that of catechins. In addition, the effect of the oral administration of phlorotannins on mice was investigated. Phlorotannins, which are oligomers of phloroglucinol, were extracted from thalli of the brown alga Ecklonia kurome and prepared by silicic acid chromatography. The bactericidal activity of polyphenols was determined using a broth microdilution method. Of the bacteria tested, Campylobacter spp. were the most susceptible to the phlorotannins. The MBCs of the crude phlorotannins, dieckol and 8,8'-bieckol (hexamers), and that of epigallocatechin gallate (EGCG) against Campylobacter jejuni were 50 mg/L, 0.03 micromol/mL and 0.03 micromol/mL, respectively. On the whole, the bactericidal effects of the phlorotannins were more pronounced than those of the catechins. The phlorotannins were as effective against MRSA as against the other bacteria tested. At twice the MBCs, all Vibrio parahaemolyticus were killed within 0.5-2 h. However, at the same concentration, catechins showed little bactericidal activity within 4 h. No effect on mice was observed with oral administration of the phlorotannins under the conditions tested.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Mar Drugs
                Mar Drugs
                marinedrugs
                Marine Drugs
                MDPI
                1660-3397
                18 June 2015
                June 2015
                : 13
                : 6
                : 3877-3891
                Affiliations
                Department of Food and Human Health Sciences, Graduate School of Human Life Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan; E-Mails: yuzu_chiffon_o712@ 123456yahoo.co.jp (H.Y.); m14hat0Q07@ 123456st.osaka-cu.ac.jp (M.G.); yuasa-i@ 123456hotmail.co.jp (I.M.-Y.)
                Author notes
                [* ]Author to whom correspondence should be addressed; E-Mail: kojima@ 123456life.osaka-cu.ac.jp ; Tel.: +81-6-6605-2865; Fax: +81-6-6605-2810.
                Article
                marinedrugs-13-03877
                10.3390/md13063877
                4483661
                26096275
                12ba408a-db5e-4389-837a-debaaf79ee5f
                © 2015 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 12 March 2015
                : 09 June 2015
                Categories
                Article

                Pharmacology & Pharmaceutical medicine
                ecklonia cava polyphenol,cytochrome p450 2e1,alcohol dehydrogenase,reactive oxygen species,cyclic amp,hepatocytes

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