Made available in DSpace on 2016-07-18T17:53:15Z (GMT). No. of bitstreams: 1 DANIELLE ALVES SILVA.pdf: 987458 bytes, checksum: 32662d7c7f3cc988d4d70c1f1d44b609 (MD5) Previous issue date: 2014-03-20 / The keratoconjunctivitis sicca ( CCS ) , or dry eye, is a common chronic inflammatory eye disease in dogs and humans , which occurs due to deficiency of production of the aqueous portion of the tear film ( quantitative deficiency ) and / or excessive tear evaporation (disability qualitative ) , mainly affecting the cornea and conjunctiva may result in blindness . Treatment consists mainly in the use of tear substitutes as the use of immunosuppressive drugs (cyclosporine , tacrolimus and pimecrolimus ) (Stevenson et al , 2012) . Some studies in medicine and veterinary showed good results in controlling the CCS with the use of essential fatty acids (AGE ) , ω -3 and ω -6 , due to its ability to produce anti -inflammatory mediators ( BARABINO et al. 2003 , . NEVES et al , 2013 ) How fish oil and flaxseed oil are sources of AGE , but with differences in composition and concentration , and to the present time there is a comparative study of the efficacy of these compounds in the treatment of CCS . The objective of this study is to compare these two important sources of ω - 3 and ω - 6 , orally , in the treatment of experimentally induced in rabbits CCS .In order to compare the efficacy in the treatment of keratoconjunctivitis sicca ( CCS ) induced experimentally in rabbits , between two sources of omega 3 and 6 orally , fish oil ( FO) and linseed oil ( OL ) . We used 28 female rabbits of New Zealand breed , divided into 4 groups of 7 animals each : Group C (control ) , Group OP ( fish oil) , OL Group ( flaxseed oil ) and Group OPL ( Oil fish oil and flaxseed ) . The animals were evaluated by Schirmer Tear Test ( TLS ) , Rose Bengal Test ( TRB ) , Fluorescein test ( TF ) , Time Tear Film Break ( TRFL ) , ocular cytology and histopathology . The results showed a significant increase in the values of TLS and TRFL in all treatment groups , but this increase was earlier in the OP group . TRB in the performance of the groups was similar. With respect to the parameter TF , negative marking was delayed in group OPL . There was a significant decrease in the number of goblet cells in the OPL group in relation to other grupos.Os results showed that OL and OP orally improve the clinical signs of CCS , however this improvement was more rapid with the OP . The combined use of oils brought no additional benefit . These results can help in the future with new oral formulations useful in the adjuvant treatment of CCS . / A ceratoconjuntivite seca (CCS), ou olho seco, é uma doença ocular inflamatória crônica frequente em cães e humanos, que ocorre devido a deficiência de produção da porção aquosa do filme lacrimal (deficiência quantitativa) e/ou pela evaporação excessiva da lágrima (deficiência qualitativa), afetando principalmente a córnea e a conjuntiva podendo evoluir para cegueira. O tratamento consiste principalmente no uso como substitutos da lágrima e o uso de imunossupressores (ciclosporina, tacrolimus e pimecrolimus) (STEVENSON et al, 2012). Alguns estudos na Medicina e na Veterinária mostraram bons resultados no controle da CCS com a utilização de ácidos graxos essenciais (AGE), ω-3 e ω-6, devido à sua capacidade de produzir mediadores anti-inflamatórios (BARABINO et al., 2003; NEVES et al., 2013) Como o óleo de peixe e o óleo de linhaça são fontes de AGE, porém com diferenças na sua composição e concentração, e até o presente momento não há um estudo comparativo da eficácia destes compostos no tratamento da CCS. O objetivo deste estudo foi comparar a eficácia no tratamento de ceratoconjuntivite seca (CCS) induzida experimentalmente em coelhos, entre duas fontes de ômega 3 e 6 por via oral, o óleo de peixe (OP) e o óleo de linhaça (OL). Foram utilizados 28 coelhos, fêmeas, da raça Nova Zelândia, divididos em 4 grupos com 7 animais cada: Grupo C (controle), Grupo OP (óleo de peixe), Grupo OL (óleo de linhaça) e Grupo OPL (óleo de peixe e óleo de linhaça). Os animais foram avaliados pelo Teste Lacrimal de Schirmer (TLS), Teste de Rosa Bengala (TRB), Teste de Fluoresceína (TF), Tempo de Ruptura do Filme Lacrimal (TRFL), citologia ocular e análise histopatológica. Os resultados demonstraram que houve aumento significativo nos valores de TLS e TRFL em todos os grupos de tratamento, porém esse aumento foi mais precoce no grupo OP. No TRB o desempenho dos grupos foi similar. Com relação ao parâmetro TF, a marcação negativa foi mais tardia no grupo OPL. Houve uma diminuição significativa no número de células caliciformes no grupo OPL em relação aos demais grupos.Os resultados demonstraram que o OP e OL por via oral melhoram os sinais clínicos da CCS, porém essa melhora foi mais rápida com o OP. O uso combinado dos óleos não trouxe benefício adicional. Esses resultados podem contribuir no futuro com novas formulações orais úteis no tratamento adjuvante da CCS.

Indiana University-Purdue University Indianapolis (IUPUI) / This dissertation studies the effects of fatty acids on prostate cancer. Prostate cancer is one of the most common malignant diseases in males in the U.S. Because of the slow progression of this disease, early intervention methods, especially, dietary fatty acid interventions are considered very important to control the disease in early stages. This study describes how the depletion of the enzyme for endogenous fatty acid synthesis, fatty acid synthase, influences the expression of enzymes that metabolize dietary fatty acids and show how dietary fatty acids affect prostate cancer protein expression and function. Fatty acid synthase is an oncoprotein overexpressed in prostate cancer and its expression is suppressed with omega-3 fatty acid treatment. This study finds that the depletion of fatty acid synthase by siRNA knockdown induces suppression of cyclooxygenase-2 and fatty acid desaturase-1. Our results also show that fish oil (omega-3 fatty acid), but not oleic acid (omega-9 fatty acid), suppresses prostate cancer cell viability. Assessment of fatty acid synthesis activity indicates that oleic acid is a more potent inhibitor than fish oil of de novo fatty acid biosynthesis. In addition, the inhibition of its activity occurs over several days while its effects on cell viability occur within 24 hours. To better understand this relationship, label free LC-MS/MS based mass spectrometry was carried out to determine global proteomic and phosphoproteomic profiles of the prostate cell line PC3, with longitudinal treatment with fish oil or oleic acid. With short-term fish oil treatment, sequestosome-1was elevated. Prolonged treatment induced downregulation of microseminoprotein, a proinflammation factor, as well as proteins in the glycolysis pathway. In the phosphoproteomics study, we confidently identified 828 phosphopeptides from 361 phosphoproteins. Quantitative comparison between fish oil or oleic acid treated groups and the untreated group suggests that the fish oil induces changes in phosphorylation of proteins involved in the pathways associated with cell viability and metabolic processes, with fish oil inducing significant decreases in the levels of phospho-PDHA1Ser232 and phospho-PDHA1Ser300 and they were accompanied by an increase in PDH activity, suggesting a role for n-3 polyunsaturated fatty acids in controlling the balance between lipid and glucose oxidation.

Die Abdeckung des Energiebedarfs ist insbesondere bei Sport- und Zuchtpferden mit einer Aufnahme hoher Stärkemengen gekoppelt. Daraus können jedoch sowohl fermentative Störungen, bedingt durch das Abfluten von Stärke in den Dickdarm, als auch metabolische Veränderungen resultieren, wobei letztere insbesondere in Form von gesteigerten Glucose- und Insulinreaktionen zum Ausdruck kommen und nicht zuletzt in der Entstehung einer Insulinresistenz münden können. In der vorliegenden Arbeit sollte der Einfluss verschiedener Nährstoffe bzw. Futtermittel in Kombination mit der Aufnahme einer stärkereichen Mahlzeit auf die Glucose- und Insulinreaktion bei gesunden Pferden überprüft werden. Für die Versuche standen insgesamt zwölf Wallache (Alter: 6 ± 4 Jahre, Gewicht: 552 ± 84 kg) zur Verfügung. Die Untersuchungen unterteilten sich in drei Fragestellungen, bei denen die jeweiligen Futtermittel randomisiert oder blockweise angeboten wurden. Als stärkereiche Mahlzeit wurde einmal täglich morgens Bruchmais in einer Dosierung von 2 g Stärke / kg KM verfüttert. Im ersten Versuchsabschnitt erhielten die Pferde verschiedene Heuzuteilungsformen (ad libitum oder restriktiv (0,6 kg / 100 kg KM)) in der Nacht vor Bruchmaisaufnahme sowie nach der Fütterung des Bruchmaises. Im zweiten Versuch wurde der Bruchmais in Kombination mit extrahierten Rohfaserquellen (0,2 g / kg KM Cellulose / Hemicellulose oder 0,1 g / kg KM Apfelpektin) oder 0,2 g Rohprotein / kg KM (in Form von Maiskleber) verfüttert. Im dritten Versuchsabschnitt erfolgte die Zulage von 0,2 ml Soja- oder Fischöl / kg KM zum Bruchmais. Je nach Fragestellung durchliefen die Pferde eine acht- bis elftägige Adaptationsphase, nach welcher sich ein- bis zweitägige Blutprobenentnahmetage anschlossen. Hierbei wurde den Pferden über einen Venenverweilkatheter bis 510 Minuten ppr. jeweils in halbstündigen Abständen Blut entnommen. Im Plasma wurden die Blutparameter Glucose (GOD-Methode) und Insulin (Radioimmunoassay) bestimmt. Es konnte gezeigt werden, dass die Zufuhr von Rohfaser in Form von Heu vor und nach Aufnahme einer stärkereichen Ration keinen wesentlichen Einfluss auf die Glucosereaktion hatte. Hierbei wurden die Glucosekonzentrationen nach zwölfstündiger Nüchterung durch die Heufütterung während der Blutentnahme nicht beeinflusst (AUCHeu restriktiv nachts, Heu restriktiv tags 132 ± 72,5 mmol x min / l, AUCHeu restriktiv nachts, Heu ad libitum tags 144 ± 114 mmol x min / l, Behandlung p > 0,05). Die Heuaufnahme während der Blutentnahme führte zu signifikant höheren Glucosekonzentrationen (Behandlung p = 0,02) wenn die Pferde vor der Bruchmaisaufnahme ad libitum Zugang zu Heu hatten (Heu ad libitum nachts, Heu ad libitum tags). Die Flächen unter den Glucosekurven wiesen jedoch keine signifikanten Unterschiede zwischen den Behandlungen auf (AUCHeu ad libitum nachts, Heu restriktiv tags 89,5 ± 50,5 mmol x min / l, AUCHeu ad libitum nachts, Heu ad libitum tags 113 ± 62,7 mmol x min / l, Behandlung p > 0,05). Die Heufütterung während der Blutentnahme ließ nach zwölfstündiger Nüchterung signifikant höhere Insulinkonzentrationen (AUCHeu restriktiv nachts, Heu ad libitum tags 5996 ± 4460 µU x min / ml) erkennen als ohne Heufütterung (AUCHeu restriktiv nachts, Heu restriktiv tags 1626 ± 1040 µU x min / ml, Behandlung p = 0,02). Die Pferde zeigten weiterhin signifikant höhere Insulinwerte, wenn nach der Fütterung des Bruchmaises Heu ad libitum aufgenommen werden konnte (AUCHeu ad libitum nachts, Heu restriktiv tags 1275 ± 845 µU x min / ml, AUCHeu ad libitum nachts, Heu ad libitum tags 3701 ± 2163 µU x min / ml, Behandlung p = 0,04). Die Zugabe extrahierter Rohfaserquellen wie Cellulose / Hemicellulose sowie Pektin zu einer definierten Maisaufnahme zeigte keinen signifikanten Einfluss auf die Glucose- (AUCBruchmais 230 ± 163 mmol x min / l, AUCCellulose / Hemicellulose 259 ± 215 mmol x min / l, AUCPektin 274 ± 106 mmol x min / l, Behandlung p > 0,05) und Insulinreaktion (AUCBruchmais 8885 ± 4024 µU x min / ml, AUCCellulose / Hemicellulose 8767 µU x min / ml, AUCPektin 10657 µU x min / ml, Behandlung p > 0,05). Durch die additive Supplementierung von Sojaöl- und Fischöl zu einer stärkehaltigen Mahlzeit ließen sich keine signifikanten Unterschiede in der Glucose- (AUCSojaöl 268 mmol x min / l, AUCFischöl 234 mmol x min / l, AUCBruchmais 257 ± 113 mmol x min / l) und Insulinreaktion (AUCSojaöl 4640 µU x min / ml, AUCFischöl 3653 µU x min / ml, AUCBruchmais 4703 ± 3365 µU x min / ml) im Vergleich zur isolierten Bruchmaisaufnahme erkennen. Die Ergänzung von Protein zu einer stärkehaltigen Mahlzeit zeigte sehr variable Reaktionen innerhalb der Einzelpferde, wobei zwei Pferde keine Reaktion und zwei Pferde erhöhte Insulinreaktionen zeigten. Unterschiede in den Glucosereaktionen wurden nicht beobachtet. Zusammenfassend ist hervorzuheben, dass die gewählten Fütterungskonzepte (Zulage extrahierter Faserquellen, Protein oder Fett) bei einer Stärkezufuhr von 2 g Stärke / kg KM keinen signifikanten Einfluss auf die Glucose- und Insulinreaktionen hatten. Dagegen führte die Fütterung von Heu ad libitum nach Aufnahme einer stärkereichen Ration zu erhöhten Insulinkonzentrationen. Um die Glucose- und Insulinreaktion nachhaltig abschwächen zu können, ist als effiziente Maßnahme eine Reduktion der aufgenommenen Stärkemenge pro Mahlzeit zu empfehlen, wohingegen die Zulage von Rohfaser und Fett bei standardisierter Stärkeaufnahme keine Abschwächung der postprandialen Glucose- und Insulinreaktion bewirkt.:INHALTSVERZEICHNIS ABKÜRZUNGSVERZEICHNIS V 1 EINLEITUNG 1 2 LITERATURÜBERSICHT 2 2.1 Einfluss verschiedener Getreidearten und deren Bearbeitungsformen auf die präcaecale Stärkeverdaulichkeit beim Pferd 2 2.2 Einfluss verschiedener Getreidearten und deren Bearbeitungsformen auf die Glucose- und Insulinreaktion beim Pferd 4 2.3 Einfluss von Rohfaser auf die Glucose- und Insulinreaktion beim Pferd 6 2.3.1 Definition und Eigenschaften von Rohfaser 6 2.3.2 Effekte löslicher Fasern auf Verdauungsvorgänge im Magen-Darm-Trakt 8 2.3.3 Effekte unlöslicher Fasern auf Verdauungsvorgänge im Magen-Darm-Trakt 10 2.3.4 Effekte von löslichen und unlöslichen Fasern auf die Glucose- und Insulinreaktion 11 2.4 Einfluss von Fett auf die Glucose- und Insulinreaktion 15 2.4.1 Klassifizierung und Eigenschaften von Fetten 15 2.4.2 Fette in der Pferdefütterung 16 2.4.3 Kurzfristige Effekte von Fettzulagen auf die Glucose- und Insulinreaktion 17 2.4.4 Langfristige Effekte von Fettzulagen auf die Glucose- und Insulinreaktion 18 2.5 Einfluss von Protein auf die Glucose- und Insulinreaktion 20 2.6 Zusammenfassung 23 3 TIERE, MATERIAL UND METHODEN 24 3.1 Versuchsplanung 24 3.2 Experimentelle Versuchsdurchführung 24 3.2.1 Einfluss von Raufutterzulagen vor und nach Aufnahme einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung I) 24 3.2.1.1 Versuchsaufbau 24 3.2.2 Effekte der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektine, Cellulose / Hemicellulose) und Protein zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung II) 28 3.2.2.1 Versuchsaufbau 28 3.2.3 Effekte unterschiedlicher Fettzulagen (Sojaöl, Fischöl) zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung III) 32 3.2.3.1 Versuchsaufbau 32 3.3 Verwendete Techniken der Futtermittelbearbeitung 34 3.3.1 Mechanische Futtermittelbearbeitung 34 3.3.2 Thermische Futtermittelbearbeitung 34 3.4 Messungen und Untersuchungsmethoden 35 3.4.1 Körpermasse der Pferde 35 3.4.2 Futteraufnahmegeschwindigkeit 35 3.4.3 Methode der Blutprobengewinnung 35 3.4.4 Untersuchungsmethoden der Futtermittel 36 3.4.4.1 Trockensubstanz und Rohnährstoffgehalte 36 3.4.4.2 Stärke 37 3.4.4.3 Zucker 37 3.4.4.4 Fettsäuren 37 3.4.5 Untersuchungsmethoden der Plasmaproben 38 3.4.5.1 Glucose 38 3.4.5.2 Insulin 38 3.4.5.3 Fettsäuren 39 3.4.6 Statistische Auswertung 39 4 ERGEBNISSE 40 4.1 Allgemeine Beobachtungen 40 4.1.1 Gesundheitszustand der Pferde 40 4.1.2 Gewichtsentwicklung der Pferde 40 4.2 Einfluss von Raufutterzulagen vor und nach Aufnahme einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung I) 42 4.2.1 Futteraufnahmeverhalten 42 4.2.2 Futteraufnahmedauer 42 4.2.3 Glucosekonzentrationen im Plasma 43 4.2.3.1 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu restriktiv tags) 43 4.2.3.2 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu ad libitum tags) 44 4.2.3.3 Vergleich der Fütterungsvarianten Heu restriktiv nachts, Heu restriktiv tags und Heu restriktiv nachts, Heu ad libitum tags 46 4.2.3.4 Effekte der Heufütterung vor Maisaufnahme ohne Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu restriktiv tags) 47 4.2.3.5 Effekte der Heufütterung vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu ad libitum tags) 48 4.2.3.6 Vergleich der Fütterungsvarianten Heu ad libitum nachts, Heu restriktiv tags und Heu ad libitum nachts, Heu ad libitum tags 50 4.2.4 Insulinkonzentrationen im Plasma 51 4.2.4.1 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu restriktiv tags) 51 4.2.4.2 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu ad libitum tags) 52 4.2.4.3 Vergleich der Fütterungsvarianten Heu restriktiv nachts, Heu restriktiv tags und Heu restriktiv nachts, Heu ad libitum tags 54 4.2.4.4 Effekte der Heufütterung vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu restriktiv tags) 55 4.2.4.5 Effekte der Heufütterung vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu ad libitum tags) 56 4.2.4.6 Vergleich der Fütterungsvarianten Heu ad libitum nachts, Heu restriktiv tags und Heu ad libitum nachts, Heu ad libitum tags 58 4.3 Effekte der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektine, Cellulose / Hemicellulose) und Protein (Maiskleber) zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung II) 59 4.3.1 Futteraufnahmeverhalten 59 4.3.2 Futteraufnahmedauer 59 4.3.3 Einfluss der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektin, Cellulose / Hemicellulose) 61 4.3.3.1 Glucosekonzentrationen im Plasma 61 4.3.3.2 Insulinkonzentrationen im Plasma 63 4.3.4 Einfluss der Zulage von Protein (Maiskleber) 65 4.3.4.1 Glucosekonzentrationen im Plasma 65 4.3.4.2 Insulinkonzentrationen im Plasma 67 4.4 Effekte unterschiedlicher Fettzulagen zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung III) 69 4.4.1 Futteraufnahmeverhalten 69 4.4.2 Futteraufnahmedauer 69 4.4.3 Fettsäuren im Plasma 69 4.4.4 Glucosekonzentrationen im Plasma 70 4.4.5 Insulinkonzentrationen im Plasma 72 4.5 Zusammenfassung der Ergebnisse 74 5 DISKUSSION 76 5.1 Kritik der Methoden 76 5.1.1 Auswahl der Versuchstiere 76 5.1.2 Versuchsdesign 76 5.1.3 Auswahl der Futtermittel 76 5.1.4 Dosierung der Futtermittel 77 5.1.4.1 Stärkegehalt in der Ration 77 5.1.4.2 Quantität der Zulagen 77 5.2 Erörterung der eigenen Ergebnisse 79 5.3 Schlussbetrachtung 89 6 ZUSAMMENFASSUNG 91 7 SUMMARY 93 8 LITERATURVERZEICHNIS 95 9 TABELLENANHANG 110 10 DANKSAGUNG 123 / In order to meet horses’ requirements of work and exercise feeding high amounts of starch is common practice. However, a high intake of starch can increase the risk of either fermentative disorders due to an overload of starch in the hindgut, or of metabolic disorders leading to exaggerated blood glucose and insulin concentrations which possibly could induce insulin resistance. The aim of this study was to investigate the effects of different nutrients in combination with the intake of a starchy meal on blood glucose and insulin concentrations in healthy horses. Twelve geldings (age: 6 ± 4 years, body weight (BW): 552 ± 84 kg) were fed the respective diets either in a randomized order or in a block design. The experiment was divided into three feeding trials. In each trial, horses received cracked corn (2 g starch / kg BW) in the morning (0800 h). In the first trial horses were fed cracked corn and grass hay in four different orders (hay ad libitum or hay restrictive (0.6 kg / 100 kg BW)), in the night before and after the intake of cracked corn. In the second trial cracked corn was mixed with extracted diatary fibers (cellulose / hemicellulose (0.2 g / kg BW) or apple pectin (0.1 g / kg BW)) or with corn gluten (2 g crude protein / kg BW). In the third trial cracked corn was fed either in combination with soybean oil (0.2 ml / kg BW) or in combination with fish oil (0.2 ml / kg BW). Each feeding period consisted of eight to eleven days of acclimatization to the different diets followed by one or two blood collection days. Blood samples were taken via a venous catheter in half-hour intervals up to 510 minutes ppr. Plasma glucose concentrations were measured by a glucose oxidase assay, and insulin levels were determined using a radioimmunoassay. There was no influence of hay feeding during blood collection after a 12 h overnight fast on plasma glucose concentrations (AUChay restriktive night, hay restriktive day 132 ± 72.5 mmol x min / l, AUChay restriktive night, hay ad libitum day 144 ± 114 mmol x min / l, treatment p > 0.05). In contrast, hay feeding during the blood collection period led to significant higher plasma glucose concentrations (treatment p = 0.02) when horses had ad libitum access to hay in the night before the intake of cracked corn. There was no significant difference in the AUC (AUChay ad libitum night, hay restriktive day 89.5 ± 50.5 mmol x min / l, AUChay ad libitum night, hay ad libitum day 113 ± 62.7 mmol x min / l, treatment p > 0.05). Furthermore, horses which were fed hay ad libitum after the intake of cracked corn demonstrated significant higher insulin concentrations (AUChay restriktive night, hay ad libitum day 5996 ± 4460 µU x min / ml) than horses without hay feeding (AUChay restriktive night, hay restriktive day 1626 ± 1040 µU x min / ml, treatment p = 0.02). In case of ad libitum hay feeding during the night significant higher insulin concentrations were found in horses which had ad libitum access to hay after intake of cracked corn when compared to horses without hay feeding during blood collection (AUChay ad libitum night, hay restriktive day 1275 ± 845 µU x min / ml, AUChay ad libitum night, hay ad libitum day 3701 ± 2163 µU x min / ml, treatment p = 0.04). The addition of extracted cellulose / hemicellulose and extracted apple pectin did not change the glycaemic (AUCcracked corn 230 ± 163 mmol x min / l, AUCcellulose / hemicellulose 259 ± 215 mmol x min / l, AUCpectin 274 ± 106 mmol x min / l, treatment p > 0,05) and insulinaemic responses (AUCcracked corn 8885 ± 4024 µU x min / ml, AUCcellulose / hemicellulose 8767 µU x min / ml, AUCpectin 10657 µU x min / ml, treatment p > 0.05) in comparison to isolated intake of cracked corn. There were no differences in postprandial glucose (AUCsoybean oil 268 mmol x min / l, AUCfish oil 234 mmol x min / l, AUCcracked corn 257 ± 113 mmol x min / l, treatment p > 0,05) and insulin responses (AUCsoybean oil 4640 µU x min / ml, AUCfish oil 3653 µU x min / ml, AUCcracked corn 4703 ± 3365 µU x min / ml, treatment p > 0.05) between feeding either cracked corn alone or in combination with soybean oil or fish oil. Protein supplementation of a starchy meal resulted in highly individual insulin responses. Two horses showed higher insulin concentrations after addition of protein to the corn meal, whereas two horses showed no reaction to the respective diet. There were no differences in the glycaemic responses. In conclusion, the results demonstrated no influence of the applied feeding strategies including supplementation of a starchy meal (2 g / kg BW) by extracted dietary fibers, protein or fat on postprandial glucose and insulin responses. In contrast, hay feeding increased the plasma insulin concentrations after the intake of cracked corn. To attenuate the glycaemic and insulinaemic responses in the horse efficiently, a reduction of starch intake per meal to quantities below 1 g starch per kg BW and meal is recommended, whereas feeding strategies like the addition of fiber or fat to a standardised starch intake do not improve glucose metabolism in healthy horses.:INHALTSVERZEICHNIS ABKÜRZUNGSVERZEICHNIS V 1 EINLEITUNG 1 2 LITERATURÜBERSICHT 2 2.1 Einfluss verschiedener Getreidearten und deren Bearbeitungsformen auf die präcaecale Stärkeverdaulichkeit beim Pferd 2 2.2 Einfluss verschiedener Getreidearten und deren Bearbeitungsformen auf die Glucose- und Insulinreaktion beim Pferd 4 2.3 Einfluss von Rohfaser auf die Glucose- und Insulinreaktion beim Pferd 6 2.3.1 Definition und Eigenschaften von Rohfaser 6 2.3.2 Effekte löslicher Fasern auf Verdauungsvorgänge im Magen-Darm-Trakt 8 2.3.3 Effekte unlöslicher Fasern auf Verdauungsvorgänge im Magen-Darm-Trakt 10 2.3.4 Effekte von löslichen und unlöslichen Fasern auf die Glucose- und Insulinreaktion 11 2.4 Einfluss von Fett auf die Glucose- und Insulinreaktion 15 2.4.1 Klassifizierung und Eigenschaften von Fetten 15 2.4.2 Fette in der Pferdefütterung 16 2.4.3 Kurzfristige Effekte von Fettzulagen auf die Glucose- und Insulinreaktion 17 2.4.4 Langfristige Effekte von Fettzulagen auf die Glucose- und Insulinreaktion 18 2.5 Einfluss von Protein auf die Glucose- und Insulinreaktion 20 2.6 Zusammenfassung 23 3 TIERE, MATERIAL UND METHODEN 24 3.1 Versuchsplanung 24 3.2 Experimentelle Versuchsdurchführung 24 3.2.1 Einfluss von Raufutterzulagen vor und nach Aufnahme einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung I) 24 3.2.1.1 Versuchsaufbau 24 3.2.2 Effekte der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektine, Cellulose / Hemicellulose) und Protein zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung II) 28 3.2.2.1 Versuchsaufbau 28 3.2.3 Effekte unterschiedlicher Fettzulagen (Sojaöl, Fischöl) zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung III) 32 3.2.3.1 Versuchsaufbau 32 3.3 Verwendete Techniken der Futtermittelbearbeitung 34 3.3.1 Mechanische Futtermittelbearbeitung 34 3.3.2 Thermische Futtermittelbearbeitung 34 3.4 Messungen und Untersuchungsmethoden 35 3.4.1 Körpermasse der Pferde 35 3.4.2 Futteraufnahmegeschwindigkeit 35 3.4.3 Methode der Blutprobengewinnung 35 3.4.4 Untersuchungsmethoden der Futtermittel 36 3.4.4.1 Trockensubstanz und Rohnährstoffgehalte 36 3.4.4.2 Stärke 37 3.4.4.3 Zucker 37 3.4.4.4 Fettsäuren 37 3.4.5 Untersuchungsmethoden der Plasmaproben 38 3.4.5.1 Glucose 38 3.4.5.2 Insulin 38 3.4.5.3 Fettsäuren 39 3.4.6 Statistische Auswertung 39 4 ERGEBNISSE 40 4.1 Allgemeine Beobachtungen 40 4.1.1 Gesundheitszustand der Pferde 40 4.1.2 Gewichtsentwicklung der Pferde 40 4.2 Einfluss von Raufutterzulagen vor und nach Aufnahme einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung I) 42 4.2.1 Futteraufnahmeverhalten 42 4.2.2 Futteraufnahmedauer 42 4.2.3 Glucosekonzentrationen im Plasma 43 4.2.3.1 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu restriktiv tags) 43 4.2.3.2 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu ad libitum tags) 44 4.2.3.3 Vergleich der Fütterungsvarianten Heu restriktiv nachts, Heu restriktiv tags und Heu restriktiv nachts, Heu ad libitum tags 46 4.2.3.4 Effekte der Heufütterung vor Maisaufnahme ohne Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu restriktiv tags) 47 4.2.3.5 Effekte der Heufütterung vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu ad libitum tags) 48 4.2.3.6 Vergleich der Fütterungsvarianten Heu ad libitum nachts, Heu restriktiv tags und Heu ad libitum nachts, Heu ad libitum tags 50 4.2.4 Insulinkonzentrationen im Plasma 51 4.2.4.1 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu restriktiv tags) 51 4.2.4.2 Effekte einer zwölfstündigen Nüchterungsphase vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu restriktiv nachts, Heu ad libitum tags) 52 4.2.4.3 Vergleich der Fütterungsvarianten Heu restriktiv nachts, Heu restriktiv tags und Heu restriktiv nachts, Heu ad libitum tags 54 4.2.4.4 Effekte der Heufütterung vor Maisaufnahme, ohne Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu restriktiv tags) 55 4.2.4.5 Effekte der Heufütterung vor Maisaufnahme in Kombination mit Heufütterung während der Blutentnahme (Heu ad libitum nachts, Heu ad libitum tags) 56 4.2.4.6 Vergleich der Fütterungsvarianten Heu ad libitum nachts, Heu restriktiv tags und Heu ad libitum nachts, Heu ad libitum tags 58 4.3 Effekte der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektine, Cellulose / Hemicellulose) und Protein (Maiskleber) zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung II) 59 4.3.1 Futteraufnahmeverhalten 59 4.3.2 Futteraufnahmedauer 59 4.3.3 Einfluss der Zulage unterschiedlicher extrahierter Rohfaserquellen (Pektin, Cellulose / Hemicellulose) 61 4.3.3.1 Glucosekonzentrationen im Plasma 61 4.3.3.2 Insulinkonzentrationen im Plasma 63 4.3.4 Einfluss der Zulage von Protein (Maiskleber) 65 4.3.4.1 Glucosekonzentrationen im Plasma 65 4.3.4.2 Insulinkonzentrationen im Plasma 67 4.4 Effekte unterschiedlicher Fettzulagen zu einer stärkehaltigen Mahlzeit auf die Glucose- und Insulinreaktion (Fragestellung III) 69 4.4.1 Futteraufnahmeverhalten 69 4.4.2 Futteraufnahmedauer 69 4.4.3 Fettsäuren im Plasma 69 4.4.4 Glucosekonzentrationen im Plasma 70 4.4.5 Insulinkonzentrationen im Plasma 72 4.5 Zusammenfassung der Ergebnisse 74 5 DISKUSSION 76 5.1 Kritik der Methoden 76 5.1.1 Auswahl der Versuchstiere 76 5.1.2 Versuchsdesign 76 5.1.3 Auswahl der Futtermittel 76 5.1.4 Dosierung der Futtermittel 77 5.1.4.1 Stärkegehalt in der Ration 77 5.1.4.2 Quantität der Zulagen 77 5.2 Erörterung der eigenen Ergebnisse 79 5.3 Schlussbetrachtung 89 6 ZUSAMMENFASSUNG 91 7 SUMMARY 93 8 LITERATURVERZEICHNIS 95 9 TABELLENANHANG 110 10 DANKSAGUNG 123

La presente investigación se realizó con el fin de encontrar los factores que influyen en la exportación de aceite de pescado (1540201000) a China, durante los años 2007 al 2017. Por tal motivo, se analizó todos los factores que se consideraron influyente al tema, siendo estas: las Medidas Sanitarias y Fitosanitarias, el Valor FOB y los Fenómenos Ambientales-El Niño. Debido a esto, se investigó sobre las Medidas Sanitarias y Fitosanitarias presentadas por China, para poder exportar y entrar a su país con este producto, ya que debido al Tratado de Libre Comercio, en el 2010, este tipo de medidas fueron más rigurosas, lo cual fue una barrera comercial para los exportadores porque eran nuevos requerimientos por obtener. Así también, se revisaron los valores FOB exportados a China del aceite de pescado, que vienen influenciados por la poca o la mucha producción de dicho producto, dependiendo de la temporada. Además, se analizó el impacto que causan ciertos fenómenos ambientales, tales como el Niño, que afecta directamente a la producción, a la biomasa de la anchoveta y su consecuencia con las exportaciones al mercado de China. Para la realización de esta proyecto, se utilizó el método de investigación cualitativo, tomando en cuenta información de expertos relacionado al tema de empresas exportadoras de aceite de pescado a China, y entidades como IMARPE, SANIPES y la Sociedad Nacional de Pesquería, con la finalidad de poder obtener una información más confiable para el desarrollo de la tesis, así mismo se utilizaron herramientas metodológicas como datos estadísticos de SUNAT y MINCETUR. / The present investigation was conducted in order to find the factors that influence the export of fish oil (1540201000) to China, during the years 2007 to 2017. For this reason, factors that were considered influential to the subject were analyzed, being these, the Sanitary and Phytosanitary Measures, the FOB Value and the Environmental-El Niño Phenomena. Due to this, it is investigated which are the Sanitary and Phytosanitary Measures presented by China, to be able to export and enter their country with this product, since due to the Free Trade Agreement, in 2010, this type of measures were more rigorous, which was a commercial barrier for exporters because they were new requirements to obtain. Likewise, the FOB values exported to China from fish oil were reviewed, which are influenced by the low or high production of said product, depending on the season. In addition, we analyzed the impact caused by certain environmental phenomena, such as El Niño, which affects production, anchovy biomass and its consequence with exports to the Chinese market. To carry out this project, the qualitative research method was used, taking into account information from experts related to the topic of fish oil exporting companies to China, and entities such as IMARPE, SANIPES and the National Fisheries Society, with the purpose of of being able to obtain a more reliable information for the development of the thesis, likewise, methodological tools were used as statistical data of SUNAT and MINCETUR. / Tesis

<p>Hypoxia-reoxygenation (H-R) injury is an important clinical phenomenon in patients with coronary artery disease (CAD). Endothelial injury is a critical step in the initiation and progression of atherosclerosis. Therefore, endothelial and cardiomyocyte protection has been considered an effective step in prevention and treatment of CAD.</p><p>To investigate the cardioprotective effect of tocopherols, omega-3 fatty acid [eicosapentaenoic acid (EPA)] and transforming growth factor-β₁ (TGF-β₁) during H-R, calcium tolerant myocytes isolated from adult rats were cultured and subjected to hypoxia for 24 hrs followed by reoxygenation of 3 hrs. All strategies, including tocopherol preparations, EPA and TGF-β₁, showed attenuation of H-R-induced myocyte injury indicated by reduction of lactate dehydrogenase (LDH) release. Both a-tocopherol and a mixed- tocopherols (α-, γ-, and δ-) decreased the effects of H-R on iNOS expression and SOD activity in cultured myocytes. The mixed-tocopherols was more potent than a-tocopherol alone. EPA inhibited H-R-induced lipid peroxidation, MMP-1 expression and p38MAPK phosphorylation. TGF-β₁ blocked the increase in iNOS and PKB phosphorylation as well as the decrease in eNOS expression in cultured myocytes exposed to H-R.</p><p> To further investigate the protective effect of omega-3 fatty acids [docosahexaenoic acid (DHA) and EPA] and TGF-β₁, the cultured endothelial cells were exposed to oxidant injury mediated by oxidized low-density lipoprotein (ox-LDL). Ox-LDL markedly reduced TGF-β₁ release, increased the expression of TGF-β₁ receptors, upregulated the expression of adhesion molecules, P-selectin and ICAM-1, enhanced the adhesion of monocytes to endothelial cells, and decreased protein kinase B (PKB) activation. Both DHA and EPA blocked these effects of ox-LDL on endothelial cells. Exogenous recombinant TGF-β₁ also ameliorated ox-LDL-induced expression of adhesion molecules and monocytes adhesion, which were blocked by antibodies to the TGF-β₁ type 2, but not to the type 3 receptor.</p><p>These observations provide mechanistic insights into H-R and oxidant injury and tissue protection by three different strategies.</p>

Hypoxia-reoxygenation (H-R) injury is an important clinical phenomenon in patients with coronary artery disease (CAD). Endothelial injury is a critical step in the initiation and progression of atherosclerosis. Therefore, endothelial and cardiomyocyte protection has been considered an effective step in prevention and treatment of CAD. To investigate the cardioprotective effect of tocopherols, omega-3 fatty acid [eicosapentaenoic acid (EPA)] and transforming growth factor-β1 (TGF-β1) during H-R, calcium tolerant myocytes isolated from adult rats were cultured and subjected to hypoxia for 24 hrs followed by reoxygenation of 3 hrs. All strategies, including tocopherol preparations, EPA and TGF-β1, showed attenuation of H-R-induced myocyte injury indicated by reduction of lactate dehydrogenase (LDH) release. Both a-tocopherol and a mixed- tocopherols (α-, γ-, and δ-) decreased the effects of H-R on iNOS expression and SOD activity in cultured myocytes. The mixed-tocopherols was more potent than a-tocopherol alone. EPA inhibited H-R-induced lipid peroxidation, MMP-1 expression and p38MAPK phosphorylation. TGF-β1 blocked the increase in iNOS and PKB phosphorylation as well as the decrease in eNOS expression in cultured myocytes exposed to H-R. To further investigate the protective effect of omega-3 fatty acids [docosahexaenoic acid (DHA) and EPA] and TGF-β1, the cultured endothelial cells were exposed to oxidant injury mediated by oxidized low-density lipoprotein (ox-LDL). Ox-LDL markedly reduced TGF-β1 release, increased the expression of TGF-β1 receptors, upregulated the expression of adhesion molecules, P-selectin and ICAM-1, enhanced the adhesion of monocytes to endothelial cells, and decreased protein kinase B (PKB) activation. Both DHA and EPA blocked these effects of ox-LDL on endothelial cells. Exogenous recombinant TGF-β1 also ameliorated ox-LDL-induced expression of adhesion molecules and monocytes adhesion, which were blocked by antibodies to the TGF-β1 type 2, but not to the type 3 receptor. These observations provide mechanistic insights into H-R and oxidant injury and tissue protection by three different strategies.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / Os ácidos graxos poli-insaturados n-3 derivados do óleo de peixe estão associados a benefícios cardiovasculares, que podem ser decorrentes da ativação da óxido nítrico sintase (NOS). Assim como as células endoteliais, os eritrócitos possuem NOS endotelial (eNOS) e induzível (iNOS) e, portanto, são capazes de sintetizar óxido nítrico (NO). O presente estudo testou a capacidade que diferentes concentrações de óleo de peixe tem de ativar a via L-arginina-NO e, em seguida, alterar os níveis de guanosina monofosfato cíclica (GMPc) em eritrócitos de camundongos alimentados com dieta hiperlipídica. Além disso, foram analisados os marcadores de estresse oxidativo nos eritrócitos, objetivando investigar a biodisponibilidade do NO. O transporte de L-arginina, avaliado através da incubação com L-[3H]-arginina, mostrou-se ativado quando da administração de dietas contendo elevadas concentrações de óleo de peixe, em comparação com as dietas contendo baixas concentrações e controle. A atividade da NOS, medida pela conversão de L-[3H]-arginina em L-[3H]-citrulina, e a expressão da eNOS também aumentaram nos animais que se alimentaram com dietas ricas em óleo de peixe. Apesar da ativação da via L-arginina-óxido nítrico observada em nossos experimentos, os níveis de GMPc intraeritrocitário não foram afetados. O dano oxidativo nos eritrócitos aumentou linearmente conforme o óleo de peixe era acrescido na dieta, sem afetar a atividade das enzimas antioxidantes. Além do endotélio, os eritrócitos contribuem para o metabolismo do NO. Desta forma, a ativação da via L-arginina-NO nessas células pode ser benéfica para saúde cardiovascular. Estudos futuros poderão investigar outros marcadores de estresse oxidativo durante o consumo de óleo de peixe para assegurar que o seu uso não resulta em efeitos prejudiciais secundários e para garantir a biodisponibilidade de NO. / The n-3 polyunsaturated fatty acids derived from fish oil are associated with cardiovascular benefits and it has been suggested that the activation of nitric oxide synthase (NOS) would be a potential mechanism responsible for its effects. Beside endothelial cells, red blood cells (RBC) possess endothelial NOS (eNOS) and inducible NOS (iNOS), and thus are capable of synthesizing their own nitric oxide (NO). The present study tested the capacity of different amounts of fish oil to activate L-arginine-NO pathway and therefore alter cyclic guanosine monophosphate (cGMP) levels in RBC from mice fed on a high fat diet. Additionally, the oxidative status in RBC was performed to investigate NO bioavailability. L-arginine transport, assessed by incubation with L-[3H]-arginine, was activated by higher doses of fish oil, compared to control diet and to lowest doses of fish oil. RBC NOS activity, measured by the conversion of L-[3H]-arginine into L-[3H]-citrulline, and eNOS expression were also enhanced by diets rich in fish oil. Despite the L-arginine-NO activation, no effect on intra RBC cGMP basal levels was seen among the groups. Oxidative damage of RBC rises linearly with increasing amounts of fish oil in the diet without affecting the activity of antioxidant enzymes. Besides endothelium, red blood cells also contribute regulating the NO bioactivity. Therefore, the activation of L-arginine-NO pathway in RBC by fish oil would be beneficial in cardiovascular health. Future studies testing other oxidant markers during dietary fish oil supplementation will be necessary to verify that its consumption does not result in detrimental secondary effects and to ensure NO bioavailability.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A modulação do tecido adiposo marrom (TAM) e do tecido adiposo branco (TAB) está associada à prevenção ou redução do ganho de massa corporal. O óleo de peixe possui diversos efeitos benéficos que podem estar relacionados a esses tecidos. Dessa forma, objetivou-se avaliar os efeitos antiobesogênicos de diferentes dietas hiperlipídicas com óleo de peixe na termogênese do TAM e na lipogênese e beta-oxidação do TAB. Para isso, foram utilizados camundongos machos C57BL/6, com três meses de idade, que foram divididos em quatro grupos experimentais: um que recebeu dieta standard-chow (SC, 10% kcal de lipídios) e outros três que receberam dieta hiperlipídica (HL, 50% kcal de lipídios). Obtivemos os grupos HL com banha de porco (HL-B), HL com banha de porco mais óleo de peixe (HL-B+Px) e HL com óleo de peixe (HL-Px). As dietas foram administradas por um período de oito semanas, sendo que a ingestão alimentar foi avaliada diariamente e a massa corporal, semanalmente. Na última semana de experimento, realizou-se a calorimetria indireta e o teste oral de tolerância à glicose. No sacrifício, a glicemia foi aferida, o sangue foi puncionado para obtenção do plasma e o TAM interescapular e o TAB epididimário foram dissecados e armazenados. A leptina, os triglicerídeos e a insulina foram mensurados no plasma. O índice de adiposidade e o HOMA-IR foram calculados. O TAM e o TAB foram avaliados por microscopia confocal e de luz. Realizou-se RT-qPCR e Western blot para avaliação de marcadores termogênicos, da captação e oxidação de ácidos graxos e glicose e de PPAR no TAM, e para a avaliação da lipogênese e beta-oxidação e de PPAR no TAB. Com relação aos resultados, o grupo HL-B apresentou ganho de massa corporal e elevação da adiposidade, associado com hipertrofia dos adipócitos, hiperleptinemia, hipertrigliceridemia, intolerância à glicose e resistência à insulina, reproduzindo um quadro de obesidade e síndrome metabólica. Por outro lado, a ingestão de óleo de peixe nos dois grupos (HL-B+Px e HL-Px) foi capaz de reduzir o ganho de massa corporal e a adiposidade, sem alterar a ingestão alimentar. Essa ingestão também aumentou o gasto energético dos animais, regularizou a leptina e os triglicerídeos plasmáticos, bem como a tolerância à glicose e a resistência à insulina. Esses efeitos foram associados ao aumento de marcadores termogênicos no TAM, bem como da captação e oxidação de ácidos graxos e glicose e da expressão de PPAR nesse tecido. No TAB, houve redução de marcadores da lipogênese e aumento de marcadores da beta-oxidação, juntamente com elevação na expressão de PPAR. Em conclusão, nossos resultados mostram que a ingestão de óleo de peixe tem efeitos antiobesogênicos em camundongos através da modulação benéfica do TAM e do TAB e pode, portanto, representar uma terapia auxiliar alternativa contra a obesidade e suas comorbidades. / Brown adipose tissue (BAT) and white adipose tissue (WAT) modulation is associated with prevention or reduction of body mass gain. Fish oil has several beneficial effects which can be related to these tissues. Thus, we aimed to evaluate the anti-obesity effects of different high-fat diets with fish oil on BAT thermogenesis and WAT lipogenesis and beta-oxidation. For this, we used 3-mo-old C57BL/6 male mice that were divided into four groups: one that received a standard-chow diet (SC, 10% kcal of lipids) and three that received a high-fat diet (HF, 50% kcal of lipids). We obtained the HF with lard group (HF-L), the HF with lard plus fish oil group (HF-L+FO), and the HF with fish oil group (HF-FO). Diets were administrated for eight weeks, and food intake was evaluated daily and the body mass, weekly. At the end of the experiment, we performed indirect calorimetry and an oral glucose tolerance test. At sacrifice, glycemia was assessed, the blood was punctured to obtain plasma and interscapular BAT and epididymal WAT were dissected and stored. Plasmatic leptin, triglycerides and insulin were analyzed. Adiposity index and HOMA-IR were calculated. BAT and WAT were evaluated through confocal and light microscopy. RT-qPCR and Western blot were performed for analyses of thermogenic markers, fatty acids and glucose uptake and oxidation, and PPAR in BAT, and lipogenesis, beta-oxidation and PPAR in WAT. In relation to the results, the HF-L group presented elevated body mass gain and adiposity, associated with adipocyte hypertrophy, hyperleptinemia, hypertriglyceridemia, glucose intolerance and insulin resistance, displaying a condition that simulates obesity and metabolic syndrome. On the contrary, fish oil intake in both groups (HF-L+FO and HF-FO) was able to reduce body mass gain and adiposity, without affecting food intake. It also increased energy expenditure, normalized plasmatic leptin and triglycerides as well as glucose tolerance and insulin resistance. These effects were associated with increases in thermogenic markers, in uptake and oxidation of fatty acids and glucose and in PPAR expression in BAT. In WAT, lipogenesis was reduced and beta-oxidation and PPAR expression were increased. In conclusion, our results demonstrated that fish oil intake has anti-obesity effects in mice through beneficial modulation of BAT and WAT and can, therefore, represent an auxiliary alternative therapy against obesity and its comorbidities.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / Os ácidos graxos poli-insaturados n-3 derivados do óleo de peixe estão associados a benefícios cardiovasculares, que podem ser decorrentes da ativação da óxido nítrico sintase (NOS). Assim como as células endoteliais, os eritrócitos possuem NOS endotelial (eNOS) e induzível (iNOS) e, portanto, são capazes de sintetizar óxido nítrico (NO). O presente estudo testou a capacidade que diferentes concentrações de óleo de peixe tem de ativar a via L-arginina-NO e, em seguida, alterar os níveis de guanosina monofosfato cíclica (GMPc) em eritrócitos de camundongos alimentados com dieta hiperlipídica. Além disso, foram analisados os marcadores de estresse oxidativo nos eritrócitos, objetivando investigar a biodisponibilidade do NO. O transporte de L-arginina, avaliado através da incubação com L-[3H]-arginina, mostrou-se ativado quando da administração de dietas contendo elevadas concentrações de óleo de peixe, em comparação com as dietas contendo baixas concentrações e controle. A atividade da NOS, medida pela conversão de L-[3H]-arginina em L-[3H]-citrulina, e a expressão da eNOS também aumentaram nos animais que se alimentaram com dietas ricas em óleo de peixe. Apesar da ativação da via L-arginina-óxido nítrico observada em nossos experimentos, os níveis de GMPc intraeritrocitário não foram afetados. O dano oxidativo nos eritrócitos aumentou linearmente conforme o óleo de peixe era acrescido na dieta, sem afetar a atividade das enzimas antioxidantes. Além do endotélio, os eritrócitos contribuem para o metabolismo do NO. Desta forma, a ativação da via L-arginina-NO nessas células pode ser benéfica para saúde cardiovascular. Estudos futuros poderão investigar outros marcadores de estresse oxidativo durante o consumo de óleo de peixe para assegurar que o seu uso não resulta em efeitos prejudiciais secundários e para garantir a biodisponibilidade de NO. / The n-3 polyunsaturated fatty acids derived from fish oil are associated with cardiovascular benefits and it has been suggested that the activation of nitric oxide synthase (NOS) would be a potential mechanism responsible for its effects. Beside endothelial cells, red blood cells (RBC) possess endothelial NOS (eNOS) and inducible NOS (iNOS), and thus are capable of synthesizing their own nitric oxide (NO). The present study tested the capacity of different amounts of fish oil to activate L-arginine-NO pathway and therefore alter cyclic guanosine monophosphate (cGMP) levels in RBC from mice fed on a high fat diet. Additionally, the oxidative status in RBC was performed to investigate NO bioavailability. L-arginine transport, assessed by incubation with L-[3H]-arginine, was activated by higher doses of fish oil, compared to control diet and to lowest doses of fish oil. RBC NOS activity, measured by the conversion of L-[3H]-arginine into L-[3H]-citrulline, and eNOS expression were also enhanced by diets rich in fish oil. Despite the L-arginine-NO activation, no effect on intra RBC cGMP basal levels was seen among the groups. Oxidative damage of RBC rises linearly with increasing amounts of fish oil in the diet without affecting the activity of antioxidant enzymes. Besides endothelium, red blood cells also contribute regulating the NO bioactivity. Therefore, the activation of L-arginine-NO pathway in RBC by fish oil would be beneficial in cardiovascular health. Future studies testing other oxidant markers during dietary fish oil supplementation will be necessary to verify that its consumption does not result in detrimental secondary effects and to ensure NO bioavailability.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A modulação do tecido adiposo marrom (TAM) e do tecido adiposo branco (TAB) está associada à prevenção ou redução do ganho de massa corporal. O óleo de peixe possui diversos efeitos benéficos que podem estar relacionados a esses tecidos. Dessa forma, objetivou-se avaliar os efeitos antiobesogênicos de diferentes dietas hiperlipídicas com óleo de peixe na termogênese do TAM e na lipogênese e beta-oxidação do TAB. Para isso, foram utilizados camundongos machos C57BL/6, com três meses de idade, que foram divididos em quatro grupos experimentais: um que recebeu dieta standard-chow (SC, 10% kcal de lipídios) e outros três que receberam dieta hiperlipídica (HL, 50% kcal de lipídios). Obtivemos os grupos HL com banha de porco (HL-B), HL com banha de porco mais óleo de peixe (HL-B+Px) e HL com óleo de peixe (HL-Px). As dietas foram administradas por um período de oito semanas, sendo que a ingestão alimentar foi avaliada diariamente e a massa corporal, semanalmente. Na última semana de experimento, realizou-se a calorimetria indireta e o teste oral de tolerância à glicose. No sacrifício, a glicemia foi aferida, o sangue foi puncionado para obtenção do plasma e o TAM interescapular e o TAB epididimário foram dissecados e armazenados. A leptina, os triglicerídeos e a insulina foram mensurados no plasma. O índice de adiposidade e o HOMA-IR foram calculados. O TAM e o TAB foram avaliados por microscopia confocal e de luz. Realizou-se RT-qPCR e Western blot para avaliação de marcadores termogênicos, da captação e oxidação de ácidos graxos e glicose e de PPAR no TAM, e para a avaliação da lipogênese e beta-oxidação e de PPAR no TAB. Com relação aos resultados, o grupo HL-B apresentou ganho de massa corporal e elevação da adiposidade, associado com hipertrofia dos adipócitos, hiperleptinemia, hipertrigliceridemia, intolerância à glicose e resistência à insulina, reproduzindo um quadro de obesidade e síndrome metabólica. Por outro lado, a ingestão de óleo de peixe nos dois grupos (HL-B+Px e HL-Px) foi capaz de reduzir o ganho de massa corporal e a adiposidade, sem alterar a ingestão alimentar. Essa ingestão também aumentou o gasto energético dos animais, regularizou a leptina e os triglicerídeos plasmáticos, bem como a tolerância à glicose e a resistência à insulina. Esses efeitos foram associados ao aumento de marcadores termogênicos no TAM, bem como da captação e oxidação de ácidos graxos e glicose e da expressão de PPAR nesse tecido. No TAB, houve redução de marcadores da lipogênese e aumento de marcadores da beta-oxidação, juntamente com elevação na expressão de PPAR. Em conclusão, nossos resultados mostram que a ingestão de óleo de peixe tem efeitos antiobesogênicos em camundongos através da modulação benéfica do TAM e do TAB e pode, portanto, representar uma terapia auxiliar alternativa contra a obesidade e suas comorbidades. / Brown adipose tissue (BAT) and white adipose tissue (WAT) modulation is associated with prevention or reduction of body mass gain. Fish oil has several beneficial effects which can be related to these tissues. Thus, we aimed to evaluate the anti-obesity effects of different high-fat diets with fish oil on BAT thermogenesis and WAT lipogenesis and beta-oxidation. For this, we used 3-mo-old C57BL/6 male mice that were divided into four groups: one that received a standard-chow diet (SC, 10% kcal of lipids) and three that received a high-fat diet (HF, 50% kcal of lipids). We obtained the HF with lard group (HF-L), the HF with lard plus fish oil group (HF-L+FO), and the HF with fish oil group (HF-FO). Diets were administrated for eight weeks, and food intake was evaluated daily and the body mass, weekly. At the end of the experiment, we performed indirect calorimetry and an oral glucose tolerance test. At sacrifice, glycemia was assessed, the blood was punctured to obtain plasma and interscapular BAT and epididymal WAT were dissected and stored. Plasmatic leptin, triglycerides and insulin were analyzed. Adiposity index and HOMA-IR were calculated. BAT and WAT were evaluated through confocal and light microscopy. RT-qPCR and Western blot were performed for analyses of thermogenic markers, fatty acids and glucose uptake and oxidation, and PPAR in BAT, and lipogenesis, beta-oxidation and PPAR in WAT. In relation to the results, the HF-L group presented elevated body mass gain and adiposity, associated with adipocyte hypertrophy, hyperleptinemia, hypertriglyceridemia, glucose intolerance and insulin resistance, displaying a condition that simulates obesity and metabolic syndrome. On the contrary, fish oil intake in both groups (HF-L+FO and HF-FO) was able to reduce body mass gain and adiposity, without affecting food intake. It also increased energy expenditure, normalized plasmatic leptin and triglycerides as well as glucose tolerance and insulin resistance. These effects were associated with increases in thermogenic markers, in uptake and oxidation of fatty acids and glucose and in PPAR expression in BAT. In WAT, lipogenesis was reduced and beta-oxidation and PPAR expression were increased. In conclusion, our results demonstrated that fish oil intake has anti-obesity effects in mice through beneficial modulation of BAT and WAT and can, therefore, represent an auxiliary alternative therapy against obesity and its comorbidities.