Investigation of sphingosin-1-phosphate-triggered matriptase activation using a rat primary hepatocyte model

Sphingosine-1-phosphate (S1P) has been reported as a matriptase activator. The aim of this study was to reveal if S1P can influence hepcidin production. Furthermore, we investigated how S1P can affect the viability and the redox status of primary hepatocytes. Rat primary hepatocytes were cultivated for 72 h and were treated with 50, 200, 1000 ng/ml S1P. Cell-free supernatants were collected every 24 h. Cell viability was tested by a colorimetric method using tetrazolium compound (MTS). The hepcidin levels in the cell-free supernatants were examined with hepcidin sandwich ELISA to determine the effect of S1P on the hepcidin-modulating ability of matriptase. In order to estimate the extent of S1P-generated oxidative stress, extracellular H2O2 measurements were performed by the use of fluorescent dye. Based on the findings, S1P treatment did not cause cell death for 72 h at concentrations up to 1000 ng/ml. S1P did not influence the extracellular H2O2 production for 72 h. The hepcidin levels were significantly suppressed in hepatocytes exposed to S1P treatment. Further studies would be needed to explore the exact mechanism of action of S1P

Szelektív matriptáz inhibitorok hatása a paracelluláris transzportfolyamatokra

A matriptáz egy olyan kettes típusú transzmembrán szerin- proteáz, amely kizárólag epithel sejteken, és epitheliális eredetű daganatokban jelenik meg. Számos szelektív matriptázgátlót találtak jelenleg is hatékonynak osteoarthritisben, influenzavírus fertőzésekben és daganatos megbetegedésekben a metasztázis-képződés gátlásában. Jelenlegi kutatásunk célja újonnan szintetizált 3-amidinofenilalanin alapvázú szelektív matriptázgátlók bélhámréteg integritására gyakorolt hatásának meghatározása, valamint a matriptáz szerepének feltérképezése a paracelluláris permeabilitás modulálásában. Kutatásunk során célul tűztük ki egy in vivo körülményekhez közelítő, a vékonybél apikális és bazolaterális kompartmentje közötti szabályozott anyagforgalom vizsgálatára alkalmas sejtmodell kifejlesztését, amelyhez egy membrán inzerten tenyésztett nem daganatos eredetű, sertés vékonybélhámsejt vonalat, az IPEC-J2 sejteket használtunk fel. A kezeléseket egy nem szelektív, szerin- proteáz inhibitor, a 4-(2-aminoetil)-benzoszulfonilfluorid (AEBSF), és több 3- amidinofenilalanin alapvázú szelektív matriptáz/TMPRSS2 inhibitor alkalmazásával végeztük

Treatment options and new perspectives in iron homeostasis disturbances Literature review

SUMMARY The authors summarize in their study the causes and the symptoms of iron toxicosis and iron deficiency anaemia and the diagnostic and treatment options of disturbances in iron homeostasis. Plasma iron level can be a reliable indica tor of iron deficiency anaemia, however, the low iron level can also be caused by chronic inflammation, infections and cancers. The iron supplementation of neonatal suckling piglets is of high veterinary importance, which is the most appropriate treatment for prevention of iron deficiency anaemia. The authors point out that iron toxicosis developed after administration of excessive iron can be compensated through parenteral deferoxamine, however other chelators applied mainly in human medicine such as deferiprone or deferasirox might be also effective in veterinary field. In this study the role of hepcidin in iron homeo stasis of the body is also discussed. The maintenance of physiological iron lev els in spite of rapid turnover of the iron suggests the regulation of hepcidin by plasma iron. In case of iron deficiency, the hepcidin amount is lowered, which then facilitates the rate of iron transport into the blood. In contrast, if the iron storage is saturated, the level of hepcidin produced by liver becomes elevated. Hepcidin oversecretion can also be induced by infection and inflammation with excessive interleukin-6 production. The authors also describe the compounds acting via hepcidin regulation which can be therapeutically beneficial in the treatment of iron homeostatic disturbances. The pharmacological intervention of the interplay between hepcidin and matriptase-2 has been one of the most recently discovered research fields which might involve the introduction of mat riptase modulators into the drug therapy of iron disorders

A rozmaringsav hatásai a haszonállatokban : Irodalmi összefoglaló = The Effects of the Rosmarinic Acid in Livestock Animals : Literature Review

The authors present in this review the in vitro and in vivo effects of rosmarinic acid, which is an ester of caffeic acid and 3,4-dihydroxy-phenyllactic acid. This phenolic acid belongs to the group of non-flavonoid polyphenols. These molecules are synthetized by plants as secondary metabolites in response to various stressful stimuli. The plants of Lamiaceae family such as common thyme, lemongrass, rosemary and sage contain high proportion of this substance. In this review rosemary and the common thyme are presented in details as these two species were usually used for in vivo experiments. Besides the rosmarinic acid, these two plants contain a lot of flavonoid and non-flavonoid type molecules in their volatile oils or in their other extracts, which have beneficial effects on human and animal health. It is widely known that the phenolic ring can act as proton and hydrogen donator, which can render this molecule antioxidant characteristic. Antitumor, antimicrobial, anti-inflammatory, gastro- and hepato-protective effects of rosmarinic acid in aglycon form were successfully proven in in vitro experiments using cell line models or tissue cultures. In the nature, however, rosmarinic acid is present in glycosylated form; therefore the results of the in vivo experiments differ from the in vitro data. Studies using farm animals did not apply rosmarinic acid only, but different plant parts or extracts, which involve rosmarinic acid among other active compounds. The application of these rosmarinic acid containing herbs could contribute to the efficient antioxidant defence mechanism in rabbits. Rosmarinic acid appeared to promote elevated growth rate in chickens and provide better quality of meat in broilers. In general, rosmarinic acid in combination with other substances could also improve cholesterol and triglyceride profiles and support antioxidant capacities against oxidative stress in livestock

The Impact of Fermented Wheat Germ Extract on Porcine Epithelial Cell Line Exposed to Deoxynivalenol and T-2 Mycotoxins

The effect of fermented wheat germ extract (FWGE) (Immunovet®) was evaluated with cotreatments with deoxynivalenol (DON) and T-2 toxin (T-2). These mycotoxins are produced by Fusarium mold species. The effects of FWGE on IPEC-J2 with DON and T-2 have not been studied until now. The IPEC-J2 porcine, nontumorigenic cell line was selected to investigate the outcome of the individually and simultaneously added compounds, as it has in vivo-like properties. The cells were treated for 24 h with the selected solutions; then, the IPEC-J2 cells were allowed to regenerate in a culture medium for an additional 24 h. In our results, DON and T-2 significantly increased the adverse impacts on cell viability and integrity of the cell monolayer. To elucidate the extent of oxidative stress, extracellular H2O2 concentrations and intracellular reactive oxygen species (ROS) were measured. FWGE appeared to be beneficial to IPEC-J2 cells given the separately and significantly decreased ROS levels. 1% and 2% FWGE could significantly reduce mycotoxin-induced oxidative stress. In conclusion, the results demonstrate that FWGE exerted protective effects to counteract the oxidative stress-provoking properties of applied fusariotoxins in the nontumorigenic IPEC-J2 cell line

Beneficial Effects of Rosmarinic Acid on IPEC-J2 Cells Exposed to the Combination of Deoxynivalenol and T-2 Toxin

Mycotoxin contamination in feedstuffs is a worldwide problem that causes serious health issues both in humans and animals, and it contributes to serious economic losses. Deoxynivalenol (DON) and T-2 toxin (T-2) are major trichothecene mycotoxins and are known to challenge mainly intestinal barrier functions. Polyphenolic rosmarinic acid (RA) appeared to have antioxidant and anti-inflammatory properties in vitro. The aim of this study was to investigate protective effects of RA against DON and T-2 or combined mycotoxin-induced intestinal damage in nontumorigenic porcine cell line, IPEC-J2. It was ascertained that simultaneous treatment of DON and T-2 (DT2: 1 μmol/L DON+5 nmol/L T−2) for 48 h and 72 h reduced transepithelial electrical resistance of cell monolayer, which was restored by 50 μmol/L RA application. It was also found that DT2 for 48 h and 72 h could induce oxidative stress and elevate interleukin-6 (IL-6) and interleukin-8 (IL-8) levels significantly, which were alleviated by the administration of RA. DT2 administration contributed to the redistribution of claudin-1; however, occludin membranous localization was not altered by combined mycotoxin treatment. In conclusion, beneficial effect of RA was exerted on DT2-deteriorated cell monolayer integrity and on the perturbated redox status of IPEC-J2 cells