O selênio exerce função marcante na qualidade do sêmen, atuando no desenvolvimento da peça intermediária, no desenvolvimento das células de Sertoli, e, como componente das selenoproteínas, principalmente a glutationa peroxidase. O selênio é utilizado comumente nas dietas animais como selenito de sódio (inorgânica) e as fontes orgânicas têm revelado diferenciada biodiponibilidade, sendo importantes para aumentar o aproveitamento pelos animais e reduzir o impacto ambiental. Assim, o experimento averiguou os efeitos do selênio orgânico nas características seminais no sêmen in natura e refrigerado a 18&ordm;C por três dias. Foram utilizados 12 cachaços divididos em três tratamentos: CONTROLE dieta formulada utilizando fonte inorgânica (0,30 ppm de selenito de sódio, n=4), INORGÂNICO dieta formulada utilizando fonte inorgânica (0,50 ppm de selenito de sódio, n=4) e ORGÂNICO - dieta formulada utilizando fonte orgânica (0,50 ppm de selênio- Sel-PlexTM, Alltech, Inc., n=4). As análises do sêmen consistiram da avaliação computadorizada da motilidade (CASA), e análises por citometria de fluxo da integridade das membranas plasmática e acrossomal, potencial de membrana mitocondrial, peroxidação lipídica, atividade da fosfolipídio hidroperóxido glutationa peroxidase. Associado a estas análises avaliou-se a concentração de selênio no plasma sangüíneo e seminal. Os dados foram submetidos à análise de variância, empregando-se o programa SAS (1999). As hipóteses testadas foram consideradas significativas quando p&lt;0,05. Na análise do sêmen in natura constatou-se que a concentração espermática, o número total de células e o número de doses foram influenciados positivamente pelo tratamento orgânico, assim como os percentuais de células normais. Os percentuais de defeitos de cabeça, peça intermediária e gota proximal foram menores nos animais suplementados com selênio levedura, contudo os defeitos de cauda do espermatozóide foram mais elevados. O defeito de colo do espermatozóide também foi reduzido pela suplementação de selênio. Porém, as integridades de membranas acrossomal e plasmática, potencial mitocondrial e peroxidação das membranas espermáticas, bem como as características de motilidade não melhoraram com a utilização de selênio levedura. A concentração de selênio no plasma sangüíneo e seminal foram maiores no grupo orgânico em comparação aos demais, assim como a atividade da enzima PHGPx. Em relação ao sêmen refrigerado averiguou-se que as integridades de membranas acrossomal e plasmática, potencial de membrana mitocondrial e peroxidação de membrana não foram influenciadas pela suplementação com selênio orgânico. Em contrapartida, as características de motilidade apresentaram diferenças significativas favoráveis ao tratamento orgânico, bem como o percentual de células normais e os defeitos de cabeça e gota proximal. O grupo orgânico apresentou maior percentual de defeitos de cauda comparativamente aos demais e a suplementação de selênio, independente da fonte reduziu os percentuais de defeitos de peça intermediária. A atividade da enzima PHGPx também foi influenciada positivamente pelo tratamento orgânico. Baseado nos resultados, efeitos positivos da suplementação de selênio orgânico (0,50 ppm) na dieta foram observados em algumas características espermáticas no sêmen in natura e refrigerado de cachaços. / The selenium plays a remarkable role on semen quality by acting in the development of middle piece, in the development of Sertoli cells and also as a component of selenoproteins, primarily of glutathione peroxidase. The selenium is usually utilized on animal diets as sodium selenite (inorganic). Organic sources have revealed a differentiated bioavailability, becoming important allies to enhance its use by the animals and reducing environmental impact. Therefore, this experiment evaluated the effects of organic selenium on seminal characteristics on raw and stored semen at 18&ordm;C for three days. Twelve boars were divided into three experimental groups: CONTROL inorganic sourced diet (0.30 ppm sodium selenite, n=4), INORGANIC inorganic sourced diet (0.50 ppm sodium selenite, n=4) and ORGANIC organic sourced diet (0.50 ppm selenium- Sel-PlexTM, Alltech, Inc., n=4). Semen analysis consisted of computerized assessment of motility (CASA) and flow cytometric analysis of plasmatic and acrosomal membranes integrity, mitochondrial membrane potential, lipidic peroxidation and phospholipid hidroperoxide glutatione peroxidase activity. Moreover, the blood and seminal plasma concentrations of selenium were evaluated. Data was submitted analysis of variance (SAS, 1999). Hypothesis tested were considered significant when p<0.05. In natura semen analysis showed that spermatic concentration, total cell number, number of doses and percentage of normal cells were positively influenced by organic treatment. Percentages of head and middle piece defects and proximal cytoplasmic droplets were decreased on animals supplemented with yeast selenium; however percentages of tail defects were increased in these animals. Neck defects were reduced by selenium supplementation. Acrosomal and plasmatic membrane integrity, mitochondrial membrane potential and spermatic membrane peroxidation, as well as motility characteristics were not enhanced with the use of yeast selenium. Selenium concentration on blood and seminal plasma were higher on organic group, similarly to PHGPx activity. Stored semen analysis showed that acrosomal and plasmatic membranes integrity, mitochondrial membrane potential and membrane peroxidation were not influenced by organic selenium supplementation. Nonetheless, motility characteristics were maintained normal with significant differences on behalf of organic treatment, likewise the percentage of normal cells, head defects and proximal droplets. The organic group presented higher percentage of tail defects when compared to the others and supplementation with selenium, independently from the source, reduced the percentages of middle piece defects. PHGPx activity was also positively influenced by organic treatment. Based on results, positives effects of dietary organic selenium supplementation (0.5 ppm) were observed on some spermatic characteristics of raw and stored boar semen.

O selênio exerce função marcante na qualidade do sêmen, atuando no desenvolvimento da peça intermediária, no desenvolvimento das células de Sertoli, e, como componente das selenoproteínas, principalmente a glutationa peroxidase. O selênio é utilizado comumente nas dietas animais como selenito de sódio (inorgânica) e as fontes orgânicas têm revelado diferenciada biodiponibilidade, sendo importantes para aumentar o aproveitamento pelos animais e reduzir o impacto ambiental. Assim, o experimento averiguou os efeitos do selênio orgânico nas características seminais no sêmen in natura e refrigerado a 18&ordm;C por três dias. Foram utilizados 12 cachaços divididos em três tratamentos: CONTROLE dieta formulada utilizando fonte inorgânica (0,30 ppm de selenito de sódio, n=4), INORGÂNICO dieta formulada utilizando fonte inorgânica (0,50 ppm de selenito de sódio, n=4) e ORGÂNICO - dieta formulada utilizando fonte orgânica (0,50 ppm de selênio- Sel-PlexTM, Alltech, Inc., n=4). As análises do sêmen consistiram da avaliação computadorizada da motilidade (CASA), e análises por citometria de fluxo da integridade das membranas plasmática e acrossomal, potencial de membrana mitocondrial, peroxidação lipídica, atividade da fosfolipídio hidroperóxido glutationa peroxidase. Associado a estas análises avaliou-se a concentração de selênio no plasma sangüíneo e seminal. Os dados foram submetidos à análise de variância, empregando-se o programa SAS (1999). As hipóteses testadas foram consideradas significativas quando p&lt;0,05. Na análise do sêmen in natura constatou-se que a concentração espermática, o número total de células e o número de doses foram influenciados positivamente pelo tratamento orgânico, assim como os percentuais de células normais. Os percentuais de defeitos de cabeça, peça intermediária e gota proximal foram menores nos animais suplementados com selênio levedura, contudo os defeitos de cauda do espermatozóide foram mais elevados. O defeito de colo do espermatozóide também foi reduzido pela suplementação de selênio. Porém, as integridades de membranas acrossomal e plasmática, potencial mitocondrial e peroxidação das membranas espermáticas, bem como as características de motilidade não melhoraram com a utilização de selênio levedura. A concentração de selênio no plasma sangüíneo e seminal foram maiores no grupo orgânico em comparação aos demais, assim como a atividade da enzima PHGPx. Em relação ao sêmen refrigerado averiguou-se que as integridades de membranas acrossomal e plasmática, potencial de membrana mitocondrial e peroxidação de membrana não foram influenciadas pela suplementação com selênio orgânico. Em contrapartida, as características de motilidade apresentaram diferenças significativas favoráveis ao tratamento orgânico, bem como o percentual de células normais e os defeitos de cabeça e gota proximal. O grupo orgânico apresentou maior percentual de defeitos de cauda comparativamente aos demais e a suplementação de selênio, independente da fonte reduziu os percentuais de defeitos de peça intermediária. A atividade da enzima PHGPx também foi influenciada positivamente pelo tratamento orgânico. Baseado nos resultados, efeitos positivos da suplementação de selênio orgânico (0,50 ppm) na dieta foram observados em algumas características espermáticas no sêmen in natura e refrigerado de cachaços. / The selenium plays a remarkable role on semen quality by acting in the development of middle piece, in the development of Sertoli cells and also as a component of selenoproteins, primarily of glutathione peroxidase. The selenium is usually utilized on animal diets as sodium selenite (inorganic). Organic sources have revealed a differentiated bioavailability, becoming important allies to enhance its use by the animals and reducing environmental impact. Therefore, this experiment evaluated the effects of organic selenium on seminal characteristics on raw and stored semen at 18&ordm;C for three days. Twelve boars were divided into three experimental groups: CONTROL inorganic sourced diet (0.30 ppm sodium selenite, n=4), INORGANIC inorganic sourced diet (0.50 ppm sodium selenite, n=4) and ORGANIC organic sourced diet (0.50 ppm selenium- Sel-PlexTM, Alltech, Inc., n=4). Semen analysis consisted of computerized assessment of motility (CASA) and flow cytometric analysis of plasmatic and acrosomal membranes integrity, mitochondrial membrane potential, lipidic peroxidation and phospholipid hidroperoxide glutatione peroxidase activity. Moreover, the blood and seminal plasma concentrations of selenium were evaluated. Data was submitted analysis of variance (SAS, 1999). Hypothesis tested were considered significant when p<0.05. In natura semen analysis showed that spermatic concentration, total cell number, number of doses and percentage of normal cells were positively influenced by organic treatment. Percentages of head and middle piece defects and proximal cytoplasmic droplets were decreased on animals supplemented with yeast selenium; however percentages of tail defects were increased in these animals. Neck defects were reduced by selenium supplementation. Acrosomal and plasmatic membrane integrity, mitochondrial membrane potential and spermatic membrane peroxidation, as well as motility characteristics were not enhanced with the use of yeast selenium. Selenium concentration on blood and seminal plasma were higher on organic group, similarly to PHGPx activity. Stored semen analysis showed that acrosomal and plasmatic membranes integrity, mitochondrial membrane potential and membrane peroxidation were not influenced by organic selenium supplementation. Nonetheless, motility characteristics were maintained normal with significant differences on behalf of organic treatment, likewise the percentage of normal cells, head defects and proximal droplets. The organic group presented higher percentage of tail defects when compared to the others and supplementation with selenium, independently from the source, reduced the percentages of middle piece defects. PHGPx activity was also positively influenced by organic treatment. Based on results, positives effects of dietary organic selenium supplementation (0.5 ppm) were observed on some spermatic characteristics of raw and stored boar semen.

Das 1817 erstmals schriftlich erwähnte Selen galt lange Zeit nur als toxisch und sogar als procancerogen, bis es 1957 von Schwarz und Foltz als essentielles Spurenelement erkannt wurde, dessen biologische Funktionen in Säugern durch Selenoproteine vermittelt werden. Die Familie der Glutathionperoxidasen nimmt hierbei eine wichtige Stellung ein. Für diese sind konkrete Funktionen und die dazugehörigen molekularen Mechanismen, welche über die von ihnen katalysierte Hydroperoxidreduktion und damit verbundene antioxidative Kapazität hinausgehen, bislang nur unzureichend beschrieben worden. <br><br> Die Funktion der gastrointestinalen Glutathionperoxidase (GI-GPx) wird als Barriere gegen eine Hydroperoxidabsorption im Gastrointestinaltrakt definiert. Neuen Erkenntnissen zufolge wird die GI-GPx aber auch in verschiedenen Tumoren verstärkt exprimiert, was weitere, bis dato unbekannte, Funktionen dieses Enzymes wahrscheinlich macht.<br> Um mögliche neue Funktionen der GI-GPx, vor allem während der Cancerogenese, abzuleiten, wurde hier die transkriptionale Regulation der GI-GPx detaillierter untersucht. Die Sequenzanalyse des humanen GI-GPx-Promotors ergab das Vorhandensein von zwei möglichen "antioxidant response elements" (ARE), bei welchen es sich um Erkennungssequenzen des Transkriptionsfaktors Nrf2 handelt. Die meisten der bekannten Nrf2-Zielgene gehören in die Gruppe der Phase-II-Enzyme und verfügen über antioxidative und/oder detoxifizierende Eigenschaften. Sowohl auf Promotorebene als auch auf mRNA- und Proteinebene konnte die Expression der GI-GPx durch typische, in der Nahrung enthaltene, Nrf2-Aktivatoren wie z.B. Sulforaphan oder Curcumin induziert werden. Eine direkte Beteiligung von Nrf2 wurde durch Cotransfektion von Nrf2 selbst bzw. von Keap1, das Nrf2 im Cytoplasma festhält, demonstriert. Somit konnte die GI-GPx eindeutig als Nrf2-Zielgen identifiziert werden. Ob sich die GI-GPx in die Gruppe der antiinflammatorischen und anticancerogenen Phase-II-Enzyme einordnen lässt, bleibt noch zu untersuchen. <br><br> Die Phospholipidhydroperoxid Glutathionperoxidase (PHGPx) nimmt aufgrund ihres breiten Substratspektrums, ihrer hohen Lipophilie und ihrer Fähigkeit, Thiole zu modifizieren, eine Sonderstellung innerhalb der Familie der Glutathionperoxidasen ein. Mit Hilfe eines PHGPx-überexprimierenden Zellmodells wurden deshalb Beeinflussungen des zellulären Redoxstatus und daraus resultierende Veränderungen in der Aktivität redoxsensitiver Transkriptionsfaktorsysteme und in der Expression atheroskleroserelevanter Adhäsionsmoleküle untersucht. Als Transkriptionsfaktoren wurden NF-kB und Nrf2 ausgewählt. Die Bindung von NF-kB an sein entsprechendes responsives Element in der DNA erfordert das Vorhandensein freier Thiole, wohingegen Nrf2 durch Thiolmodifikation von Keap1 freigesetzt wird und in den Kern transloziert. Eine erhöhte Aktivität der PHGPx resultierte in einer Erhöhung des Verhältnisses von GSH zu GSSG, andererseits aber in einer verminderten Markierbarkeit freier Proteinthiole. PHGPx-Überexpression reduzierte die IL-1-induzierte NF-kB-Aktivität, die sich in einer verminderten NF-kB-DNA-Bindefähigkeit und Transaktivierungsaktivität ausdrückte. Auch war die Proliferationsrate der Zellen vermindert. Die Expression des NF-kB-regulierten vaskulären Zelladhäsionsmoleküls, VCAM-1, war ebenfalls deutlich verringert. Umgekehrt war in PHGPx-überexprimierenden Zellen eine erhöhte Nrf2-Aktivität und Expression der Nrf2-abhängigen Hämoxygenase-1 zu verzeichnen. Letzte kann für die meisten der beobachteten Effekte verantwortlich gemacht werden.<br><br> Die hier dargestellten Ergebnisse verdeutlichen, dass eine Modifizierung von Proteinthiolen als wichtige Determinante für die Regulation der Expression und Funktion von Glutathionperoxidasen angesehen werden kann. Entgegen früheren Vermutungen, welche oxidative Vorgänge generell mit pathologischen Veränderungen assoziierten, scheint ein moderater oxidativer Stress, bedingt durch eine transiente Thiolmodifikation, durchaus günstige Auswirkungen zu haben, da, wie hier dargelegt, verschiedene, miteinander interagierende, cytoprotektive Mechanismen ausgelöst werden. Hieran wird deutlich, dass sich "antioxidative Wirkung" oder "oxidativer Stress" keineswegs nur auf "gute" oder "schlechte" Vorgänge beschränken lassen, sondern im Zusammenhang mit den beeinflussten (patho)physiologischen Prozessen und dem Ausmaß der "Störung" des physiologischen Redoxgleichgewichtes betrachtet werden müssen. / Selenium was discovered in 1817 by the Swedish chemist Berzelius and was for a long time considered as being toxic and even procarcinogenic. In 1957, however, Schwarz and Foltz realized that selenium is an essential trace element which elicits its biological functions in mammals as a structural component of selenoproteins among which the family of glutathione peroxidases plays a dominant role. Glutathione peroxidases reduce hydroperoxides to the corresponding alcohols and contribute to the antioxidative capacity of a cell. However, other functions of glutathione peroxidases and the according molecular mechanisms have hardly been described.>br><br> The gastrointestinal glutathione peroxidase (GI-GPx) is believed to build a barrier against the absorption of foodborne hydroperoxides. In addition, GI-GPx expression is increased in different tumors. This indicates further, still unknown, functions of this enzyme.<br> In order to elucidate new possible functions of GI-GPx, especially during carcinogenesis, the transcriptional regulation of GI-GPx was analyzed in more detail. An analysis of the GI-GPx promoter sequence revealed the presence of two putative "antioxidant response elements" (ARE) which are recognition sites for the transcription factor Nrf2. Most of the known Nrf2 target genes either belong to the group of phase-II detoxification enzymes or possess antioxidative and/or detoxifying properties. On promoter level as well as on mRNA- and protein level the expression of GI-GPx was induced by typical Nrf2-activating compounds such as sulforaphane or curcumin that are contained in the diet. A direct involvement of Nrf2 was demonstrated by cotransfection of Nrf2 itself or by cotransfection of Keap1 which retains Nrf2 in the cytosol. Thus, the GI-GPx gene was unequivocally identified as a new target for Nrf2. Whether GI-GPx also belongs in the category of antiinflammatory and anticarcinogenic enzymes remains to be elucidated.<br><br> The phospholipid hydroperoxide glutathione peroxidase (PHGPx) is exceptional among the glutathione peroxidases because of its broad range of substrates, its high lipophilicity, and its ability to modify protein thiols. With PHGPx-overexpressing cells, the influence of PHGPx on the cellular redox state and on resulting changes in the activity of redox-sensitive transcription factors and on the expression of proatherogenic adhesion molecules was analyzed. For this, the redox-sensitive transcription factors NF-kB and Nrf2 were chosen. NF-kB requires free thiols for being able to bind to its responsive element within the DNA, whereas Nrf2 is released from Keap1 and translocates to the nucleus upon a modification of protein thiols. PHGPx-overexpression resulted in an increase in the ratio of GSH to GSSG, in a reduced amount of intracellular protein thiols, and in a diminished proliferation rate. Furthermore, PHGPx-overexpressing cells displayed a reduced IL-1-dependent NF-kB activity as was assessed by a reduced NF-kB DNA-binding ability and activity of a NF-kB-driven reporter gene. In addition, the expression of the NF-kB-dependent vascular cell adhesion molecule (VCAM-1) was also inhibited by overexpression of PHGPx. On the other hand, PHGPx-overexpressing cells displayed an increased activity of Nrf2 that was accompanied by an increased expression of the Nrf2-dependent heme oxygenase-1. Heme oxygenase-1 most likely is responsible for most of the aforementioned effects.<br><br> The data presented here show that a modification of protein thiols can be regarded as an important determinant for the regulation and for the functions of glutathione peroxidases. In contrast to the previous assumption that oxidative processes are always linked to pathologic changes, a moderate oxidative stress seems to have beneficial effects, because it triggers different cytoprotective mechanisms. It can be concluded that the terms "antioxidative effect" or "oxidative stress" cannot simply be restricted to "good" or "bad" processes, but need to be seen in context with the modulated (patho)physiological processes and the degree of "disturbance" of the physiologic redox balance.