At the end of the 20th century it was known that cannabinoid drugs interact with two receptors, CB1 and CB2. Subsequent pharmacological studies have confirmed that there are other receptors interacting with cannabinoids. GPR55 is a transmembrane G protein coupled receptor, which together with the receptor GPR18 and GPR119 belong to a group of new cannabinoid receptors and is involved in the function of the endocannabinoid system. In addition to some of cannabinoid substances, it is stimulated primarily phospholipid lysofosfatidylinositolem. LPI-dependent signaling GPR55 plays an important role in the regulation of many physiological and pathological processes, such as pain, inflammation, cell proliferation, or endothelial function. It was found that LPI confers tolerance to ischemic brain damage and has a cytoprotective effect on the pyramidal cells. The aim of the study was to determine whether the application of five ligands induce phosphorylation of protein kinase ERK 1/2, Akt and activate the GTPase RhoA and whether activation of the receptor GPR55 has cytoprotective effect in model cell line PC12, in which hypoxic conditions were simulated by adding CoCl2. For working methods were used SDS-PAGE, Western bloting and colorimetric measurement. Pharmacological studies in recent years have shown...

Thesis (Ph.D.)--Medical College of Ohio, 2004. / "In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Medical Sciences." Major advisor: Linda A. Dokas. Document formatted into pages: iv, 150 p. Title from title page of PDF document. Bibliography: pages 44-52.

The heat shock response (HSR) is the cell's molecular reaction to protein damaging stress and is critical in the management of denatured proteins. Activation of HSF1, the master transcriptional regulator of the HSR, results in the induction of molecular chaperones called heat shock proteins (HSPs). Transcription of hsp genes is promoted by the hyperphosphorylation of HSF1, while the attenuation of the HSR is regulated by a dual mechanism involving negative feedback inhibition from HSPs and acetylation at a critical lysine residue within the DNA binding domain of HSF1, which results in a loss of affinity for DNA. SIRT1 is a NAD+-dependent histone deacetylase that has been reported to deacetylate HSF1, thus promoting stress-induced HSF1 DNA binding ability and increasing HSP expression (Westerheide, Anckar et al. 2009). While an abundance of research is aimed to investigate SIRT1 substrate regulation, the mechanism in which SIRT1 itself is regulated is less understood (Haigis and Sinclair 2010). Positive and negative modulators of SIRT1 include AROS and DBC1, respectively, and have yet to be investigated in relation to SIRT1-dependent regulation of the HSR. In addition, metabolic stress such as caloric restriction has been shown to modulate SIRT1 activity in yeast (Rahat, Maoz et al. 2011), but the effect of caloric restriction on the HSR is unknown. Using cell-based assays, we have investigated how the HSR may be controlled by factors influencing SIRT1 activity. We found that heat shock results in an increase in the cellular NAD+/NADH ratio and an increase in recruitment of SIRT1 to the hsp70 promoter. Furthermore, we found that the SIRT1 modulators, AROS and DBC1, impact hsp70 transcription, HSF1 acetylation status, and HSF1 recruitment to the hsp70 promoter. The nematode Caenorhabditis elegans is a useful model organism for testing the relationship between the HSR and metabolism, as these animals can easily be calorically-restricted via bacterial limitation and possess the mammalian SIRT1 homolog, Sir2.1. Using C. elegans, we demonstrate that caloric restriction and heat shock have a synergistic effect on the HSR in a sir2.1-dependent manner. We show that caloric restriction increases the ability of heat shock to promote thermotolerance and fitness in wild-type animals and to preserve movement in a polyglutamine toxicity neurodegenerative disease model and that this effect is dependent on sir2.1. These studies provide insight into SIRT1-dependent regulation of the HSR and the impact of metabolism on this response. We highlight the SIRT1 modulators AROS and DBC1 as two new targets available for therapeutic regulation of the HSR and add caloric restriction as another HSR activator that can synergize with heat shock.

Quercetin-3-O-glucoside (Q3G), a glycosylated derivative of quercetin, is a polyphenolic compound known to possess diverse biological activities. Its moderately hydrophilic nature is a critical factor governing the accessibility to the active sites of oxidative damages in vivo. It was hypothesized that biological activities of Q3G can be further enhanced by regioselective acylation with fatty acids which gives more lipophilicity. Q3G was acylated with six selected long chain fatty acids: stearic acid, oleic acid, linoleic acid, ?-linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), using Candida antactica lipase. The derivatives were evaluated for their potential in inhibiting lipid oxidation in food systems and human low density lipoprotein (LDL), and cytoprotection and anti-inflammatory effect in cell culture model systems. The fatty acid derivatives of Q3G possessed greater effectiveness in inhibiting lipid oxidation in oil-in-water emulsions, and better cytoprotective effect against H2O2- and cigarette smoke toxicant-induced cytotoxicity when compared to Q3G.

Au Canada, nous remarquons une prédominance du diabète de type 2 au sein des communautés autochtones. Une approche ethnobotanique est utilisée en collaboration avec la Nation Crie de Eeyou Istchee afin de déterminer quels traitements à base de plantes peuvent être utilisés pour contrer les différentes conditions qui, collectivement, forment le diabète. Les pharmacopées de deux communautés cries, soit celles de Waskaganish et de Nemaska, ont été établies puis comparées à celles de étudiées antérieurement : communautés Whapmagoostui et Mistissini. Malgré les différences géographiques de ces groupes, leurs utilisations sont majoritairement semblables, avec pour seule exception le contraste entre les communautés de Nemaska et de Whapmagoostui. De plus, nous avons complété l’évaluation du taux cytoprotecteur des aiguilles, de l’écorce et des cônes de l’épinette noire (Picea mariana). Les extraits provenant de tous les organes des plantes démontrent une protection qui dépend de la concentration. La réponse spécifique d’organes peut varier selon l’habitat; ainsi, les plantes poussant dans les tourbières ou dans les forêts, sur le littoral ou à des terres l’intérieur démontrent des différences quant à leur efficacité. Bref, l’écorce démontre une relation dose-effet plus forte dans la forêt littorale, tandis que les aiguilles n’indiquent pas de changements significatifs selon leur environnement de croissance. La bioactivité observée démontre une corrélation avec le contenu phénolique et non avec l’activité de l’agent antioxydant. Ces résultats contribuent à péciser les activités antidiabétiques des plantes de la forêt boréale canadienne, telles qu’identifiées au niveau cellulaire par les guérisseurs Cries. / In Canada there is an overwhelming prevalence of type II diabetes in First Nations communities. Here an ethnobotanical approach has been used in cooperation with the Cree Nation of Eeyou Istchee to focus on finding plant based treatments for the conditions which collectively make up the symptoms of diabetes. The pharmacopoeias of two Cree communities (Waskaganish and Nemaska) are elucidated then compared with previously studied populations (Whapmagoostui and Mistissini). Despite differences in north-south east-west geography, plant ranking and use matrices were similar with the exception of Nemaska/Whapmagoostui. We have also completed the evaluation of Black spruce (Picea mariana) needle, bark and cone cytoprotectivity. Extracts from all organs exhibited concentration-dependent protection. Organ-specific response was habitat and growth environment dependent with plants grown either in bog or forest habitats in coastal or inland environments exhibiting differences in efficacy. Observed bioactivity correlated with total phenolic content but not with antioxidant activity. Together, these results contributed to the understanding of antidiabetic activity of Canadian boreal forest plants identified by the Cree of Eeyou Istchee healers at the cellular level.

Au Canada, nous remarquons une prédominance du diabète de type 2 au sein des communautés autochtones. Une approche ethnobotanique est utilisée en collaboration avec la Nation Crie de Eeyou Istchee afin de déterminer quels traitements à base de plantes peuvent être utilisés pour contrer les différentes conditions qui, collectivement, forment le diabète. Les pharmacopées de deux communautés cries, soit celles de Waskaganish et de Nemaska, ont été établies puis comparées à celles de étudiées antérieurement : communautés Whapmagoostui et Mistissini. Malgré les différences géographiques de ces groupes, leurs utilisations sont majoritairement semblables, avec pour seule exception le contraste entre les communautés de Nemaska et de Whapmagoostui. De plus, nous avons complété l’évaluation du taux cytoprotecteur des aiguilles, de l’écorce et des cônes de l’épinette noire (Picea mariana). Les extraits provenant de tous les organes des plantes démontrent une protection qui dépend de la concentration. La réponse spécifique d’organes peut varier selon l’habitat; ainsi, les plantes poussant dans les tourbières ou dans les forêts, sur le littoral ou à des terres l’intérieur démontrent des différences quant à leur efficacité. Bref, l’écorce démontre une relation dose-effet plus forte dans la forêt littorale, tandis que les aiguilles n’indiquent pas de changements significatifs selon leur environnement de croissance. La bioactivité observée démontre une corrélation avec le contenu phénolique et non avec l’activité de l’agent antioxydant. Ces résultats contribuent à péciser les activités antidiabétiques des plantes de la forêt boréale canadienne, telles qu’identifiées au niveau cellulaire par les guérisseurs Cries. / In Canada there is an overwhelming prevalence of type II diabetes in First Nations communities. Here an ethnobotanical approach has been used in cooperation with the Cree Nation of Eeyou Istchee to focus on finding plant based treatments for the conditions which collectively make up the symptoms of diabetes. The pharmacopoeias of two Cree communities (Waskaganish and Nemaska) are elucidated then compared with previously studied populations (Whapmagoostui and Mistissini). Despite differences in north-south east-west geography, plant ranking and use matrices were similar with the exception of Nemaska/Whapmagoostui. We have also completed the evaluation of Black spruce (Picea mariana) needle, bark and cone cytoprotectivity. Extracts from all organs exhibited concentration-dependent protection. Organ-specific response was habitat and growth environment dependent with plants grown either in bog or forest habitats in coastal or inland environments exhibiting differences in efficacy. Observed bioactivity correlated with total phenolic content but not with antioxidant activity. Together, these results contributed to the understanding of antidiabetic activity of Canadian boreal forest plants identified by the Cree of Eeyou Istchee healers at the cellular level.

A cisplatina (cis-diaminodicloroplatina II) é um efetivo agente anticâncer, porém seu uso clínico é altamente limitado, predominantemente devido à sua nefrotoxicidade. Muitos estudos têm demonstrado que a cisplatina causa disfunção mitocondrial em células epiteliais renais devido à ação de espécies reativas de oxigênio tais como ânions superóxido e radicais hidroxila. A proteção seletiva das mitocôndrias renais contra espécies reativas de oxigênio geradas pela cisplatina é fundamental na quimioterapia de pacientes com câncer. Vários estudos têm sugerido que o carvedilol é capaz de proteger contra a toxicidade mitocondrial cardíaca induzida pelo quimioterápico doxorubicina. Assim, no presente estudo investigou-se o potencial protetor deste fármaco contra a toxicidade mitocondrial renal induzida pela cisplatina, bem como os mecanismos moleculares envolvidos nesta proteção. Foram estudados 4 grupos (n=6, cada) de ratos Wistar machos tratados da seguinte forma: (i) Grupo controle: uma injeção intraperitoneal (i.p.) de DMSO (0,2mL/200g, i.p.) imediatamente antes da injeção de solução salina isotônica (2 ml/200g, i.p.) e posteriormente uma injeção diária de DMSO (0,2mL/200g, i.p.) em dois dias consecutivos; (ii) Grupo cisplatina (CISP): uma injeção de cisplatina (10 mg/kg, i.p.); (iii) Grupo carvedilol (CV): uma injeção de carvedilol (1 mg/kg, i.p.), seguida de uma injeção diária de carvedilol em dois dias consecutivos (1 mg/Kg, i.p) e (iv) Grupo carvedilol + cisplatina (CV+CISP): uma injeção de carvedilol (1mg/kg, i.p.), imediatamente antes da injeção de cisplatina (10 mg/Kg, i.p.) seguida de uma injeção diária de carvedilol nos dois dias seguintes (1 mg/Kg, i.p.). Os animais foram sacrificados 72 horas após o início do tratamento. O grupo CV+CISP apresentou uma significativa redução na lesão renal, marcada pela diminuição da concentração de uréia e creatinina plasmáticas, quando comparado ao grupo CISP. A avaliação da função mitocondrial comprovou o efeito protetor do carvedilol contra a toxicidade mitocondrial renal da cisplatina através da significativa melhora nos valores da (i) RCR, (ii) do consumo de oxigênio no estado 3 e (iii) da razão ADP/O. Além disso, no grupo CV+CISP o potencial de membrana mitocondrial e a captação de cálcio mitocondrial foram preservados. Adicionalmente, a redução da oxidação do NADPH, da cardiolipina, da glutationa e das proteínas sulfidrilas, bem como a redução na formação de MDA no grupo CV+CISP sugerem efeito protetor do carvedilol contra o estresse oxidativo mitocondrial. O grupo CV+CISP também apresentou menor ativação da caspase-3, o que sugere menor indução de apoptose. Os grupos CISP e CV+CISP apresentaram concentrações semelhantes de platina na suspensão mitocondrial renal, indicando que o mecanismo de proteção do carvedilol provavelmente não envolve a formação de complexos e a subseqüente inativação da cisplatina. Os resultados do presente estudo são promissores, pois o carvedilol é um fármaco de uso seguro e já estabelecido na clínica e a comprovação do seu efeito protetor contribuirá para o desenvolvimento de novas estratégias de prevenção dos danos nefrotóxicos da cisplatina. / Cisplatin (cis-diamminedichloridoplatinum II) is an affective anticancer agent; however its clinical use is highly limited, predominantly due to its nephrotoxicity. Many studies have shown that cisplatin cause mitochondrial dysfunction in renal epithelial cells due to the generation of reactive oxygen species, such as superoxide anions and hydroxyl radicals. The selective protection of the renal mitochondria against the reactive oxygen species generated by cisplatin is of critical importance in the chemotherapy of cancer patients. Some studies have suggested that carvedilol can protect against the cardiac mitochondrial toxicity induced by the chemotherapeutic agent doxorubicin. Therefore, in the present study we investigated the protective effect of carvedilol against renal mitochondrial toxicity, as well as the molecular mechanisms involved in this protection. We studied 4 groups (n=6, each) of male Wistar rats treated as follows: (i) Control group: one injection of DMSO (0,2 mL/200g body weight, i.p.), intraperitoneal injection (i.p.), immediately before the injection of isotonic saline solution (2mL/200g body weight) followed by one injection of DMSO (0,2 mL/200g body weight, i.p.) in the two following days; (ii) Cisplatin group (CISP): one injection of cisplatin (10 mg/kg body weight, i.p.); (iii) Carvedilol group (CV): one injection of carvedilol (CV) (1mg/kg body weight, i.p.) in three consecutive days and (iv) Carvedilol + Cisplatin group (CV+CISP): one injection of carvedilol (1mg/kg, body weight, i.p.) immediately before the injection of cisplatin (10mg/kg, body weight, i.p.), followed by one injection of carvedilol (1mg/kg, body weight, i.p.) in the two following days. Animals were killed 72h after the beginning of the treatment. CV+CISP group presented a significantly reduced renal injury, marked by the decrease of urea and creatinine plasmatic levels, as compared to the CISP group. The evaluation of the mitochondrial function showed the protective effect of carvedilol against the renal mitochondria toxicity induced by cisplatin, as demonstrated by the improvement in the values of (i) RCR; (ii) the oxygen consumption on state 3 respiration and ADP/O ratio. Besides that, in the CV+CISP group the mitochondrial membrane potential and the mitochondrial calcium uptake were preserved. Additionally, the lower oxidation of NADPH, cardiolipin, glutathione and sulfhydryl proteins, as well as the lower values of MDA in the CV+CISP group, suggests a protective effect of carvedilol against the mitochondrial oxidative stress. CV+CISP group also presented lower values of caspase 3, which suggests lower induction of apoptosis. The groups CISP and CV+CISP presented a similar platinum concentration in the mitochondrial suspension, which indicates that the protective mechanism of carvedilol probably does not involve complex formation with cisplatin and its ensuing inactivation. The present results are promising, since carvedilol is a safe drug, which is currently used in the clinical practice and the evidence of its protective effect will contribute to the development of new strategies to prevent the nephrotoxic damage of cisplatin.

As plantas do gênero Copaifera mais bem conhecidas como \"copaibeira\" são árvores tropicais, reconhecidas pela produção de uma oleorresina extraída do tronco, a qual vem sendo empregada por mais de 500 anos na medicina tradicional popular. Entre as espécies mais abundantes no Brasil destaca-se Copaifera langsdorffii L., que nas suas folhas têm um grande número de compostos fenólicos com potencial atividade biológica como antioxidante. Este estudo teve como finalidade isolar compostos das classes de flavonoides e galoilquínicos presentes nas folhas de C. langsdorffii e investigar seus possíveis efeitos citoprotetores frente à intoxicação com metais pesados. Foi obtido o extrato hidroalcoólico das folhas, e foi submetido a partições líquido-líquido. A fração em acetato de etila foi fracionada através de cromatografia contracorrente de alta velocidade (HSCCC), e a fração em n-butanol por cromatografia de exclusão em gel Sephadex. Foram isolados e identificados por RMN, dois flavonoides (quercitrina e afzelina) da fração em acetato de etila e dez derivados galoilquínicos da fração n-butanólica. Os flavonoides e dois derivados galoilquínicos (AGQ3 e AGQ4), foram avaliados em ensaios de citotoxidade com o protocolo de alamar blue nas linhagens celulares de hepatocarcinoma humano (HepG2) e feocromocitoma da medula suprarrenal de rato (PC12), em três diferentes concentrações (1, 10 e 100 ?M) por 24 h. A segurança do composto AGQ4 também foi avaliada através do ensaio de eficiência clonogênica contra a linhagem de pulmão de hamster chinês (V79). Após foram feitos ensaios de associação realizando pré-tratamentos de 1 hora com os compostos, para posterior exposição a doses tóxicas dos metais. Os ensaios de citotoxidade demostraram que nenhuma das concentrações avaliadas dos quatro compostos afetou a viabilidade das linhagens celulares. O ensaio de eficiência clonogênica mostrou que apenas uma concentração de 3,8 mM consegue reduzir significativamente a viabilidade na linhagem V79, mostrando o AGQ4 como um composto seguro para o uso in vitro. Os ensaios de associação mostraram que os quatro compostos têm efeito citoprotetor significativo na linhagem HepG2, contra mercúrio e chumbo a exceção do AGQ3 que em concentração de 100 ?M não mostrou proteger contra o chumbo. Por outro lado, os compostos não foram ativos contra o mercúrio na linhagem PC12, mostrando efeito contrário, pois os tratamentos metal-composto diminuíram a viabilidade em comparação com os tratamentos só com o metal. Apenas nas concentrações de 1 e 10 ?M se evidenciou efeito protetor dos compostos contra o chumbo. Em geral também foi observado um efeito de menor atividade na maior concentração do composto, que pode estar relacionado ao efeito pró-tóxico que alguns polifenóis apresentam em concentrações maiores. Tanto os flavonoides quanto os derivados galoilquínicos isolados de C. langsdorffii se mostraram com potencial na citoproteção contra a exposição ao chumbo e ao mercúrio / The Copaifera plants, better known as \"copaibeira\", are tropical trees recognized by the production of an oleoresin extracted from the tree trunk, which has been used for more than 500 years in traditional folk medicine. Among the most abundant species in Brazil is Copaifera langsdorffii L., which in its leaves have a large number of phenolic compounds with potential biological activity as antioxidant. Then, this study aimed to isolate compounds of the classes of flavonoids and galloylquinic derivatives present in the leaves of C. langsdorffii and to investigate their possible cytoprotective effects against heavy metal poisoning. The hydroalcoholic extract of the leaves was obtained and submitted to liquid-liquid partitions. The ethyl acetate fraction was subjected to high-speed countercurrent chromatography (HSCCC), and the fraction in n-butanol was subjected to Sephadex gel exclusion chromatography. Two flavonoids (quercitrin and afzelin) from the ethyl acetate fraction and ten galloylquinic derivatives of n-butanolic fraction were isolated and identified by NMR, using semi-preparative high performance liquid chromatography (CLAE-UV). Flavonoids and two galloylquinic acids (AGQ3 and AGQ4) were evaluated in alamar blue cytotoxicity assays in HepG2 cell lines (human hepatocarcinoma cells) and PC12 (rat adrenal pheochromocytoma cells), in three different concentrations (1, 10 and 100 ?M) for 24 h. The safety of compound AGQ4 was also assessed by the clonogenic efficiency assay against V79 (Chinese hamster lung) cell line. Once was proved that compound concentrations did not affect cell viability, association assays were performed by one hour pre-treatments of the compounds for further exposure to toxic doses of the metals. Cytotoxicity assays demonstrated that none of the evaluated concentrations of the four compounds affected the viability of the cell lines. The clonogenic efficiency assay showed that only a concentration of 3.8 mM of AGQ4 can significantly reduce viability in the normal cell line, showing the galloylquinic derivative as a highly safe compound for in vitro use. The association assays showed that the four compounds have a significant cytoprotective effect for the HepG2 cell line against Hg and Pb, except for AGQ3 which at 100 ?M concentration did not show protection against lead. Also the compounds did not show activity against Hg in the cell line PC12 and, in addition, they gave the opposite effect by decreasing cell viability in comparison with the treatments with only the metal. Only the concentrations of 1 and 10 ?M showed a protective effect of the compounds against lead. In general, a lower activity effect was also observed at the higher concentrations of the compounds, which may be related to the protoxic effect of some phenolic compounds. Both flavonoids and galloylquinic acid derivative compounds showed potential on the cytoprotection for exposure to lead and methylmercury.

INTRODUÇÃO E HIPÓTESE: A epinefrina é um potente vasoconstritor com efeitos inotrópico e arritmogênico, é utilizada em protocolos de reanimação cardiopulmonar e como fármaco de primeira escolha em alguns casos de choque. Contudo, o seu uso pode ser seguido por lesões do miocárdio e disfunção cardíaca. Modelos experimentais têm mostrado efeitos cardioprotetores do fentanil por meio de mecanismos antiarrítmicos e anti-isquêmicos. O objetivo deste estudo foi avaliar o efeito cardioprotetor do fentanil em suínos expostos a altas doses de epinefrina. MÉTODOS: Após aprovação do comitê de ética institucional, 26 porcos Large White e Landrace foram alocados aleatoriamente em três grupos: grupo fentanil (n = 10), no qual os porcos receberam 20 ug/kg de fentanil 5 minutos antes de 5 doses de 20 ug/kg de epinefrina, as quais foram intercaladas por intervalos de 5 minutos entre cada dose; grupo salina (n = 10), no qual os porcos receberam solução salina volume-equivalente ao fentanil 5 minutos antes das 5 doses de epinefrina e grupo Sham (n = 6), que não recebeu fentanil ou epinefrina. Foram coletadas variáveis hemodinâmicas, ecocardiográficas, gasométricas e marcadores cardíacos durante as 6 horas de experimento. Ao final do estudo, o coração e os pulmões dos porcos foram removidos para análise por microscopia óptica, microscopia eletrônica e imuno-histoquímica (caspase-3). Os dados foram analisados usando equações de estimação generalizadas (GEE) e a significância estatística foi estabelecida em p < 0,05. RESULTADOS: Os níveis de troponina-I entre os grupos foram inicialmente equivalentes. Ao final do experimento, foi observado menor nível de troponina-I no grupo fentanil, em comparação com o grupo salina (1,91 ± 1,47 versus 5,44 ± 5,35 ng.ml-1, p = 0,019). Adicionalmente, a microscopia eletrônica e a imunohistoquímica demonstraram menor lesão miocárdica no grupo fentanil. Não houve diferença significativa entre o grupo fentanil e o salina para as variáveis hemodinâmicas, ecocardiográficas e gasométricas. CONCLUSÃO: O fentanil promove cardioproteção aos efeitos de altas doses de epinefrina sem prejudicar o efeito hemodinâmico da mesma / INTRODUCTION AND HYPOTHESIS: Epinephrine is a powerful vasopressor with inotropic and arrhythmogenic effects that is used in cardiopulmonary resuscitation protocols and as first choice drug in some cases of shock. However, its use could be followed by myocardial injury and dysfunction. Experimental models have shown cardioprotective effects of fentanyl through antiarrhythmic and anti-ischaemic mechanisms. The objective of this study was to evaluate the cardioprotective effect of fentanyl on myocardial function in swine exposed to high doses of epinephrine. METHODS: After institutional ethics committee approval, twenty-six Large White and Landrace pigs were allocated randomly into three groups: Fentanyl group (n=10), which received 20ug/kg of fentanyl five minutes before five doses of 20ug/kg of epinephrine interspersed with 5 minute intervals between each dose; Saline group (n=10), which received saline in a volume-equivalent manner of fentanyl five minutes before 20ug/kg of epinephrine doses; and Sham group (n=6), which did not receive fentanyl nor epinephrine. We assessed hemodynamics, transesophageal echocardiography, cardiac markers, and gasometry for 6 h. At the end of the experiment, the heart and lungs were removed for analysis by optical and electron microscopy and immunohistochemical (Caspase-3) assay. Data was analyzed using generalized estimating equations (GEE) and statistical significance was assumed at p < 0.05. RESULTS: Troponin levels among the groups were initially equivalent. Fentanyl group showed lower levels of troponin at the end of the sixth hour compared to the saline group (1.91 ± 1.47 vs. 5.44 ± 5.35 ng.mL-1, p=0.019). There were no significantly difference between fentanyl and saline group for hemodynamic, echocardiographic and gasometrical data. Transmission electron microscopy and immunohistochemistry also showed less myocardial injury in the fentanyl group. CONCLUSION: We concluded that fentanyl promotes effective cardioprotection to high-dose epinephrine without blunting the hemodynamic effect of epinephrine

Made available in DSpace on 2017-07-21T19:59:50Z (GMT). No. of bitstreams: 1 Simoni Marineia Braatz.pdf: 10395189 bytes, checksum: 4c6c59c43b05d7cdd5092ae73a0d7737 (MD5) Previous issue date: 2010-02-22 / Surinam cherry leaves are popularly used as an alternative treatment for diabetes mellitus. They are rich in phenolic compounds which have antioxidant properties. Besides, it is well known that reactive species influence in the progression of diabetes. The aim of this study was to evaluate the antioxidant, cytotoxic and cytoprotective activity of the crude leaf extracts of Eugenia uniflora L. and some major components of the extracts, quercetin (QE) and myricetin (MI); and the effect of these substances on the functionality of RINm5f cells. The methodologies performed were: Folin- Ciocalteau method, MTT reduction, DPPH method, hematoxylin-eosin stain, radioimmunoassay and Western Blot analyses. The total phenolic content obtained for the extracts was 440.27 g/mL and 37.83Yg/mL catechin equivalent/mg of aqueous extract (AE) and ethanol extract (EE), respectively. In the DPPH essay, the IC50 of AE was 321.50 g/mL, MI was 152.63 g/mL and QE was 110.278 g/mL. The EE didn’t show significant antioxidant activity. The AE and the EE are not cytotoxic in the concentrations between 12.5 - 75 g/mL and 31.25 - 500Yg/mL, respectively. QE and MI are not cytotoxic in the concentrations between 3.13 – .25 g/mL and 3.13 – 12.5 g/mL, respectively. Most of the compounds didn’t interfere in insulin secretion, except for EE, which increased at least 3 times insulin secretion and MI which decreased insulin secretion in its lower concentration tested. Hydrogen peroxide (H2O2) induced a reduction of 33% of cell viability in acute assay and 45% in chronic assay. The AE had a cytoprotective effect mainly in the chronic assay and in its higher concentrations. The EE had a similar effect only when chronically tested. QE showed a dose-dependent result (6.5 > 3.13 g/mL) in the acute assay and prevented 100% of the damage caused by H2O2 in the chronic assay. One possible cytoprotective mechanism for these compounds and extracts is the inhibition of the activation of NF-κB. Besides that, H2O2 altered intercellular adhesion with intense modification of cellular morphology. The cells presented a round shape, with the formation of folds in the membrane and reduction of cytoplasm basophily. Both extracts and isolated compounds also prevented changes in morphology caused by H2O2, suggesting prevention of lipid peroxidation. There was no variation in the expression of the proteins superoxide dismutase (SOD), Bax, disulfide isomerase (PDI) and GRP94 (glucoseregulated protein), however, the increased protein content of GRP78 was an indicative of reticular stress. These results are an evidence that Eugenia uniflora L. can be used as an alternative treatment for diabetes mellitus. / Folhas de pitanga são popularmente utilizadas no tratamento alternativo do diabetes mellitus. Portanto, o objetivo deste trabalho foi avaliar a atividade antioxidante, citotóxica e citoprotetora dos extratos brutos das folhas de Eugenia uniflora L. e de compostos encontrados nos extratos: quercetina (QE) e miricetina (MI), além do efeito destes sobre a funcionalidade de células RINm5f. As metodologias utilizadas foram: Folin-Ciocalteau, redução do MTT, captura do DPPH, coloração por hematoxilina e eosina, radioimunoensaio e Western Blot. O conteúdo total de fenóis obtido nos extratos foi 440,27 g/mL e 37,83 g/mL equivalentes de catequina/mg de extrato aquoso (EA) e alcoólico (EE), respectivamente. No ensaio de captura do DPPH, o EA apresentou um IC50 de 321,50 g/mL, a MI de 152,63 g/mL e a QE de 110,278 g/mL. O EE não apresentou significativa atividade antioxidante. O EA e o EE não apresentaram citotoxicidade nas concentrações entre 12,5 - 75 g/mL e 31,25 - 500 g/mL, respectivamente. Já a QE e MI são mais citotóxicas, apresentando viabilidade celular superior a 90% entre 3,13 - 6,25 g/mL e 3,13 - 12,5 g/mL, respectivamente. Quanto à funcionalidade celular, o EA e a QE não interferiram na secreção de insulina. A MI causou redução na secreção de insulina na concentração de 3,13 g/mL enquanto o EE aumentou mais de 3 vezes a secreção deste hormônio. O peróxido de hidrogênio (H2O2) reduziu, em média, 33% da viabilidade das células no ensaio agudo e 45% no ensaio crônico. O EA promoveu citoproteção principalmente no ensaio crônico e nas maiores concentrações testadas. Já o EE apresentou citoproteção apenas quando incubado cronicamente. A QE apresentou resultado dose-dependente (6,5 > 3,13 g/mL) no ensaio agudo e preveniu 100% dos danos causados pelo H2O2 no ensaio crônico. Um possível mecanismo de ação relacionado com a proteção dos extratos e compostos isolados é o bloqueio da ativação do NF-κB pelo H2O2. Além disso, o H2O2 alterou a adesão intercelular com intensa modificação na morfologia destas. As células apresentaram formato mais arredondado, com formação de pregas na membrana e redução da basofilia citoplasmática. Os extratos e compostos isolados preveniram alterações na morfologia causadas por H2O2, sugerindo prevenção da peroxidação lipídica. Não houve variação na concentração protéica da superóxido dismutase (SOD), BAX, dissulfito isomerase (PDI) e GRP94 (proteína regulada por glicose), entretanto houve aumetno da concentração da proteína GRP78 indicando estresse reticular. Os resultados apresentados são um indicativo de que a E. uniflora L. possa ser usada no tratamento alternativo do diabetes mellitus.