Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: This study describes: - the characterization and comparison of the enzymatic activity of both Angora and ovine 3βHSD expressed in non-steroidogenic COS-1 cells. The apparent Km and Vmax values for the metabolism of PREG, 17-OHPREG and DHEA were determined; - the characterization of steroid metabolites produced by COS-1 cells coexpressing either Angora or ovine 3βHSD together with Angora CYP17, in the presence and absence of overexpressed Cyt-b5, following the metabolism of PREG and 17-OHPREG. 3βHSD was identified as an additional factor in causing hypocortisolism in the South African Angora goat; - the influence of Cyt-b5 on the enzymatic activity of both Angora and ovine 3βHSD coexpressed in non-steroidogenic COS-1 cells; - the influence of purified ovine live Cyt-b5 and anti-Cyt-b5 IgG on adrenal microsomal 3βHSD activity. Cyt-b5 was shown to specifically augment 3βHSD activity which represents the first documentation of such augmentation in any species; - the overexpression and purification of Angora 3βHSD using a baculovirus expression system coupled with a detergent based enzyme purification method; - the characterization of both substrate and co-factor kinetics for the individual dehydrogenase and isomerase activities of purified 3βHSD, in the presence and absence of purified ovine liver Cyt-b5. Cyt-b5 was shown to increase the affinity of 3βHSD towards NAD+ during the dehydrogenase reaction whilst having no significant influence on the isomerase reaction. This represents the first documentation of Cyt-b5 influencing co-factor binding in any member of the -ydroxysteroid dehydrogenases; - the FRET analysis of COS-1 cells coexpressing 3βHSD-eCFP and Cyt-b5-eYFP fusion proteins, suggesting an allosteric interaction between 3βHSD and Cyt-b5. / AFRIKAANSE OPSOMMING: Hierdie studie beskryf: - die karakterisering en vergelyking van die ensiematiese aktiwiteit van beide Angora en skaap 3βHSD, wat uitgedruk was in nie-steroïed genererende COS-1 selle. Die Km en Vmax waardes tydens die metabolisme van PREG, 17-OHPREG en DHEA was bepaal; - die karakterisering van steroïed metaboliete gegenereer deur COS-1 selle wat Angora of skaap 3βHSD uitdruk saam met Angora CYP17, in die aanwesigheid of afwesigheid van sitochroom b5, na die metaboliseering van PREG en 17-OHPREG. 3βHSD was geïdentifiseer as ‘n bydraende faktor in die oorsaak van hipokortisolisme in die Suid-Afrikaanse Angorabok; - die invloed van sitochroom b5 op die ensiematiese aktiwiteit van beide Angora en skaap 3βHSD wat saam uitgedruk was in nie-steroïed genererende COS-1 selle; - die invloed van gesuiwerde skaap lewer sitochroom b5 en sitochroom b5 teenstof op mikrosomale 3βHSD aktiwiteit. Dit is getoon dat sitochroom b5 die aktiwiteit van 3βHSD spesifiek verhoog. Hierdie studie verteenwoordig die eerste dokumentasie van so ‘n verhoging in enige spesie; - die uitdrukking en suiwering van Angora 3βHSD deur middel van ‘n bakulo-virus sisteem gekoppel aan ‘n detergent gebaseerde ensiem suiwerings metode; - die karakterisering van beide substraat en ko-faktor kinetika vir die afsonderlike dehidrogenase en isomerase aktiwiteite van gesuiwerde 3βHSD, in die aanwesigheid of afwesigheid van gesuiwerde sitochroom b5. Dit is getoon dat sitochroom b5 die affiniteit van 3βHSD teenoor NAD+ tydens die dehidrogenase reaksie verhoog sonder om ‘n beduidende invloed op die isomerase reaksie te hê. Hierdie studie verteenwoordig die eerste dokumentasie van sitochroom b5 wat ko-faktor binding beïnvloed in enige lid van die hidroksisteroïed dehidrogenase familie van ensieme; - die analise van FRET sein in COS-1 selle wat beide 3βHSD-eCFP en Cyt-b5- eYFP fusie proteïene uitdruk. Die resultate stel voor dat sitochroom b5 3βHSD aktiwiteit beïnvloed deur middel van ‘n allosteriese meganisme.

Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: This study describes: • the influence of a methanolic extract of unfermented Rooibos and five major Rooibos flavonoids, aspalathin, nothofagin, rutin, orientin and vitexin, on the activities of key adrenal steroidogenic enzymes - cytochrome P450 17β- hydroxylase/17,20-lyase (CYP17A1), 3β-hydroxysteroid dehydrogenase • the development of a novel UPLC-MS/MS method for the separation and quantification of 21 adrenal steroid metabolites; • the influence of Rooibos and aforementioned flavonoids on adrenal steroid hormone production in H295R cells - a human adrenal carcinoma cell line expressing the enzymes catalysing the production of mineralocorticoids, glucocorticoids and adrenal androgens, assayed under both basal (normal) and forskolin-stimulated (stressed) conditions; • the influence of Rooibos on the inter-conversion between cortisol and cortisone by 11βHSD1 and 11βHSD2 expressed in CHO-K1 cells; • the influence of Rooibos consumption on circulating steroid hormone levels and ratios in male Wistar rats; • the influence of Rooibos consumption on circulating steroid hormone levels and ratios in male and female human test subjects at risk for developing cardiovascular disease. (3βHSD2), cytochrome P450 21-hydroxylase (CYP21A2) and cytochrome P450 11β-hydroxylase (CYP11B1), expressed in non-steroidogenic COS-1 cells; / AFRIKAANSE OPSOMMING: Hierdie studie beskryf: • die invloed van metanoliese ekstrakte van ongefermenteerde Rooibos en vyf van die hoof flavonoïedverbindings in Rooibos, aspalatien, notofagien, rutien, oriëntien en viteksien, op die aktiwiteite van ensieme wat steroïedbiosintese in die bynier kataliseer – sitochroom P450 17α-hidroksilase/17,20-liase (CYP17A1), 3β-hidroksisteroïed dehidrogenase (3βHSD2), sitochroom P450 21-hidroksilase (CYP21A2) en sitochroom P450 11β-hidroksilase (CYP11B1), uitgedruk in nie-steroïed produserende COS-1 selle; • die ontwikkeling van ‘n geskikte UPLC-MS/MS metode vir die skeiding en kwantifisering van 21 steroïedmetaboliete in die bynier; • die invloed van Rooibos en die bg. flavonoïede op steroïedproduksie in H295R selle – ‘n menslike bynier kanker sellyn gekenmerk deur die ekspressie van die steroidogeniese ensieme wat die produksie van mineralokortikoïede, glukokortikoïede en bynierandrogene kataliseer, geanaliseer onder beide basale (normale) en forskoliengestimuleerde (gestresde) kondisies; • die invloed van Rooibos op die omeenskakeling tussen kortisol en kortisoon deur 11βHSD1 and 11βHSD2 in CHO-K1 selle; • die invloed van Rooibosinname op vlakke van sirkulerende steroïed hormone en relatiewe verhoudings in die bloed van manlike Wistarrotte; • die invloed van Rooibosinname op sirkulerende steroïed hormoon vlakke en relatiewe verhoudings in die bloed van mans en vrouens met ‘n hoë risiko vir die ontwikkeling van kardiovaskulêre siektes.

published_or_final_version / Obstetrics and Gynaecology / Doctoral / Doctor of Philosophy

Estrogens, such as the biologically active 17-b estradiol (E2) have many actions in the male and female brain. Not only does E2 regulate reproductive behavior in adults, it organizes and activates the brains of younger animals in a sex-specific manner. In addition, many human studies have shown E2 to provide protection against a variety of neurological disorders, including stoke. These studies have been controversial and depend largely on the type and timing of hormone replacement. Animal studies are much less controversial and clearly demonstrate a neuroprotective role for E2 following ischemic brain injury. Because much of E2 neuroprotection requires sex steroid hormone receptors, it is essential to understand expression patterns of these receptors. For the current studies, I evaluated estrogen receptor alpha (ER α), estrogen receptor beta (ER β) and androgen receptor (AR) expression in the cortex. It is known that these receptors change in expression at several times in an animal’s life span including during early postnatal development and following ischemic brain injury. Here I used an in vitro cortical explant model to further examine how these receptors change both during development and following injury. This in vitro model is important because it provides a way to investigate changes in receptor expression pattern in the cortex without input from other brain regions. In addition to characterizing this model, I also evaluated the contribution of E2 to changes in receptor expression and on cell death following injury in the explants. To begin to decipher mechanisms for E2 mediated neuroprotection, I added antagonist for each of the receptors before and after injury. In each these experiments, I also examined potential sex differences by separating the female and male brains before I cultured the explants. Overall, these experiments showed that cortical explants are a good in vitro model. Here we found that E2 was protective in female, but not male cortical explants following injury. However, the exact mechanisms of E2-mediated neuroprotection are still to be deciphered.

This thesis investigates the biological activity of selected non-steroidal analogues of sex steroid hormones by examining two different effects of analogues on endogenous sex hormone activity. Non-steroidal analogues of sex hormones were synthesised to study their biological interactions with a sex steroid receptor and a sex steroid metabolising enzyme. Chapter One introduces the steroid hormones and their physiology, which leads to a review of the mechanisms by which steroids exert their effects. Their implication in disease is discussed, with particular emphasis on the sex steroids. As the biological activity of steroids is related to their chemical structure, the important features of steroid structure are identified, including the cyclopentanoperhydrophenanthrene nucleus, arrangement of ring substituents and ring junction conformation. The concept of non-steroidal analogues of steroids is introduced, and the harmful or beneficial effects analogues have on endogenous steroid activity are considered. Alteration of steroid activity and its consequences are focussed on two main areas; the potential adverse effects of environmental chemicals which mimic sex steroid activity, and the use of non-steroidal analogues in medicinal chemistry for treating sex steroid related disease. Chapter Two describes an investigation into the 17β-estradiol mimicking activity of non-steroidal analogues. Exogenous chemicals that mimic estradiol are of concern as they may alter endogenous estradiol activity and disrupt endocrine systems. Firstly, an introduction to the field of research concerned with environmental chemicals that mimic steroid hormones is given. The interaction of xenoestrogens with the estrogen receptor is described, as are the methods available for assessing the estrogen mimicking activity of xenoestrogens. The concern for insecticides mimicking estrogen activity is described by reviewing reported activities of insecticides, which leads into a discussion of work carried out as part of this thesis. Metabolites of the pyrethroid insecticides permethrin and cypermethrin, 2.14, 2.15, and 2.16 were synthesised while others were commercially obtained. The interaction of pyrethroid insecticide metabolites with the human estrogen receptor expressed in recombinant yeast (Saccharomyces cerevisiae) was studied, following the establishment and validation of the assay. Metabolites 2.11, 2.12, and 2.14 were found to weakly stimulate estrogen receptor-mediated estradiol responsive gene expression in the yeast assay (105 less active than 17β-estradiol). Since the activity of the metabolites using the yeast assay was greater than for the parent compounds, metabolic pathways need to be considered when assessing the impact of exposure to environmental estrogens. The low estrogenic activity suggests these compounds are not individually contributing significantly to the xenoestrogenic impact on humans, but will add to total xenoestrogen exposure. Chapter Three describes the inhibition of a sex steroid metabolising enzyme, steroid 5a-reductase, by novel non-steroidal compounds. Inhibitors of this enzyme are potentially useful therapeutic agents for regulating the activity of an androgen in prostate disorders. A review of the literature on non-steroidal inhibition of 5a-reductase identified three key structural features known to enhance inhibitor potency; ring substitution, position and nature of ring unsaturation and angular methyl group presence. These features were taken into account in the design of inhibitors synthesised in this thesis (3.55-3.57, 3.59, 3.61, 3.62, 3.110 and 3.111). Inhibitors consisting of non-steroidal 5- or 1-aryl pyridone scaffolds were synthesised to investigate SAR for 4'-substituents. The 5-aryl 1-methyl-2-pyridone/piperidone scaffold of compounds 3.55-3.57 and 3.59 was constructed by Suzuki cross coupling methodology, while the 1-aryl 2-methyl 2,3-dihydro-4-pyridone scaffold of 3.61 and 3.62 was constructed by aza Diels-Alder methodology. Long carbon chain olefin containing tethers 3.107 and 3.108 were synthesised for conjugation to inhibitor 3.57 by cross metathesis to give conjugates 3.110 and 3.111. Compounds 3.55-3.57, 3.59, 3.61, 3.62, 3.110 and 3.111 inhibited the type 1 5a-reductase isozyme expressed by HEK-I cells, with activities comparable to those of related literature compounds. The 1-aryl 2,3-dihydro-4-pyridone 3.62 inhibited both the type 1 and 2 isozymes (expressed by HEK-II cells) of 5a-reductase. The presence of bulky hydrophobic groups (benzoyl, long chain tethers) at the 4' position enhanced the potency of type 1 inhibition by 5-aryl pyridone type compounds in comparison to N,N-diisopropyl- and N-allylacetamide groups. This information provides further understanding of SAR within and across different classes of non-steroidal inhibitors of steroid 5a-reductase towards improved drug design.

Les activités d'élevage sont responsables de la dissémination dans l'environnement d'un grand nombre de polluants, dont les perturbateurs endocriniens (PE) qui affectent le système endocrinien des organismes vivants. La disruption endocrinienne se fait par la liaison des PE sur des récepteurs nucléaires spécifiques dont le récepteur aux strogènes ERα, le récepteur à la dioxine AhR, le récepteur X aux pregnanes PXR, le récepteur aux androgènes AR et le récepteur régulant la prolifération des peroxysomes, PPARγ. Les hormones stéroïdes sont particulièrement actives sur le récepteur ERα. Elles sont naturellement produites par les animaux et concentrées dans les effluents d'élevage épandus sur les sols agricoles. Ce travail de thèse s'est focalisé sur l'étude du devenir des hormones dans des systèmes de gestion des lisiers porcins, à échelle réelle ou contrôliée, en conditions anaérobies ou aérobies. Au préalable, la méthodologie pour l'analyse des hormones dans le lisier a été validée. Les résultats montrent que les hormones sont principalement contenues dans la fraction solide; dans cette fraction les mesures d'activité strogénique (ERα) ont révélé une activité non explicable par les concentrations en hormones, suggérant la présence d'autres composés ayant une activité sur ERα. La quantification des nonylphénols, actifs sur ERα n'a pas permis d'expliquer cette activité. L'étude d'autres voies de perturbation endocrinienne via l'activation des récepteurs AhR, PXR, PPARγ et AR a permis de détecter une forte activité sur le récepteur AhR. L'analyse chimique de cette fraction a montré l'absence de dioxines et d'hydrocarbures aromatiques polycycliques, suggérant l'implication d'autres PE qui restent à être identifiés. Dans les systèmes de traitement sous conditions réelles ou contrôlées, les conditions aérobies sont plus efficaces pour éliminer les hormones et les activités strogéniques et « dioxin-like » associées, bien que cette dernière semble plus récalcitrante. / Livestock activities are responsible for the release of a wide range of pollutants in the environment. Among these pollutants, the endocrine disruptors (ED) can affect the endocrine functions of living organism. The endocrine disruption occurs via the binding of ED to specific nuclear receptors such as the estrogen receptor ERα, dioxin receptor AhR, pregnane-X-receptor (xenobiotics), androgen receptor AR and peroxysome proliferator-activated receptor γ, PPARγ. Steroid hormones are particularly active on ERα receptor. Animals naturally produce high amounts of hormones which are concentrated in manure used for land spreading. This study deals with the fate of hormones in swine waste management facilities at full or pilot scale, under both anaerobic and aerobic conditions. Firstly, the analysis of hormones in such a complex matrix was validated. Hormones are mainly present in the solid fraction of the manure. At the same time, estrogenic activity measurements showed that in this solid fraction, the estrogenic activity cannot be explained by hormones; which suggest the presence of other compounds active on ERα. Nonylphenols concentrations cannot explain this additional activity. The study of the other pathways of endocrine disruption through the activation of AhR, PXR, PPARγ, and AR allowed to measure high AhR activity in the solid fraction of manure. The chemical analysis of this fraction did not show the presence of dioxins and polycyclic aromatic hydrocarbons traditionally involved in such kind of activity; it suggests the presence of other AhR ligands that remain to be identified. In swine manure treatment systems, aerobic conditions were more efficient to remove hormones and the associated estrogenic and AhR activities, even if this last seemed to be more recalcitrant.

The thesis focuses on selected, yet unsolved question of the role of less common steroids and SHBG as a junction of three endocrine systems. Answering these questions may help to understand the complex mechanism of action of these hormones on the human organism. This thesis is based on five author and co-written studies mostly published in foreign scientific journals. In the case of studies of metabolites 16α-OH-DHEA and 7-oxo-DHEA (Zamrazilová et al., 2007; Kazihnitková et al., 2007) were focused on the development of appropriate methodological approach because recently used methods showed a low sensitivity and specificity. We developed and statistically evaluated new RIA methods which are rapid, sensitive and inexpensive. Both of them can be used in other research even in routine practice. New methods were also used to determine the metabolites in a statistically significant sample of healthy human population. In another study (Zamrazilová et al., 2010) we examined the relationships of selected steroid metabolites and SHBG in patients with CAH. We assumed that SHBG may act as a non-steroidal laboratory parameter reflecting the effectiveness of substitution therapy in these patients. Our assumption was not confirmed. We observed lower levels of SHBG which at least in women reflect effects of...

La chimiorésistance des cancers est caractérisée par une résistance pléïotropique à de multiples médicaments. Ce mécanisme est en partie causé par la surexpression des transporteurs à «ATP binding cassette» (Pgp, MRP1, BCRP…). Les inhibiteurs connus de ces transporteurs comme la cyclosporine A, le vérapamil et le RU486 sont toxiques à doses élevées. Dans cette étude, de nombreux dérivés stéroïdiens synthétisés au laboratoire à base de progestérone ou d’acides biliaires ont été évalués pour leur capacité à inhiber les transporteurs ABC et plus spécifiquement les fonctions de transport par la Pgp ou la BCRP. Plusieurs de ces dérivés synthétisés se sont avérés capables de restaurer complètement la sensibilité des cellules résistantes d’une manière plus importante que la cyclosporine A in vitro. De plus, le meilleur des nos dérivés testés s’est avéré capable in vivo de diminuer significativement la progression tumorale de xénogreffe sur les souris et d’augmenter la durée de survie des souris. Cette étude a ainsi permis d’ouvrir la voie au développement de nouveaux dérivés stéroïdiens peu toxiques ayant la capacité d’inhiber le phénotype MDR et de restaurer la sensibilité des cellules cancéreuses vis-à-vis des agents chimiothérapeutiques utilisés, avec un perspective d’application clinique / The chemoresistance of cancer is characterized by a pleitropic resistance to multiple drugs. This mechanism is partly caused by the overexpression of “ATP Binding Cassette” transporters (Pgp, MRP1, BCRP…). Known inhibitors of these transporters such as cyclosporin A, verapamil and RU486 rather toxic at high doses. In this study, many steroid derivatives synthesized in the laboratory (from progesterone or bile acid precursor) to bind and inhibit ABC transporters, specifically the transport by Pgp or BCRP. Several of these synthetic derivatives were able to completely restore the sensivity of the resistant cells compared to cysclosporine A in vitro. In addition, the efficient of these derivatives could, in vivo significantly reduce tumor growth of xenografts on mice and increase survival time of mice. This study opens a route for development of new steroid derivatives with low toxicity, having the ability to reverse the MDR phenotype and restore sensitivity of cancer cells to the chemotherapeutic agents use, with a perspective at clinical use

A exibição do comportamento vocal em muitas espécies de anfíbios anuros é relacionada aos níveis de hormônios esteroides gonadais e interrenais. Esses hormônios poderiam mediar a relação entre intensidade de sinais e imunidade, pois estão envolvidos no desenvolvimento das características sexuais secundárias, comportamento de corte e mobilização de reservas energéticas durante a atividade reprodutiva, enquanto apresentam também efeitos imunomoduladores. Nesse sentido, o objetivo deste trabalho foi explorar as relações entre comportamento reprodutivo, imunidade e níveis plasmáticos de testosterona e corticosterona em machos do sapo do semiárido brasileiro, Rhinella granulosa, em atividade reprodutiva e após manipulação hormonal. A precipitação foi o principal determinante ambiental para o aumento dos níveis de testosterona e corticosterona circulantes em machos de R. granulosa, estimulando o comportamento de vocalização. As relações fisiológicas encontradas indicam que os altos níveis plasmáticos de testosterona nos primeiros dias após a chuva devem promover o início do turno vocal, porém a corticosterona deve modular o esforço de vocalização. De forma geral, a exacerbação do comportamento vocal de R. granulosa tem efeitos negativos sobre a imunocompetência, porém alguns indivíduos que apresentam maiores concentrações plasmáticas de corticosterona apresentam concomitantemente alto esforço vocal e alta imunidade. De acordo, a aplicação transdérmica de corticosterona promoveu elevação aguda dos níveis plasmáticos deste glicocorticoide, bem como um aumento da função imune. Assim, apesar de a atenção principal ser comumente colocada no papel da testosterona na mediação de sinais honestos, nossos resultados corroboram a importância da corticosterona na mediação da expressão do comportamento de corte e imunocompetência em machos R. granulosa / In most anuran species, vocal behavior is usually related to plasmatic levels of gonadal and interrenal steroid hormones. These hormones could mediate the relationship between signal intensity and immunocompetence, since they are involved in the development of sexual traits, courtship behavior and energetic mobilization during reproductive activity and also exert immunomodulatory effects. The aim of this study was to explore relations among vocal behavior, immunity and plasmatic levels of testosterone and corticosterone in males of a toad species from Brazilian semi-arid, Rhinella granulosa, during reproductive activity and after hormonal manipulation. Precipitation was the main environmental determinant to increase circulating levels of testosterone and corticosterone in R. granulosa males, stimulating vocal behavior. Physiological relations indicate that high circulating testosterone levels in the days right after rain may promote the initiation of vocal behavior, while corticosterone may modulate vocal effort. In general, exacerbation of vocal behavior in R. granulosa has negative effects on immunocompetence, however, some individuals with higher plasmatic levels of corticosterone can present both high vocal effort and good immunocompetence. Corroborating this interpretation, acute increase in glucocorticoid plasmatic levels after hormonal manipulation was accompanied by raised immune function. Despite the emphasis on androgens influence on sexual selected male traits, our results corroborate greater influence of corticosterone in the mediation of expression of courtship behavior and immunocompetence in R. granulosa

INTRODUÇÃO : Distrofia Muscular de Duchenne (DMD) é uma doença genética, o que acarreta alteração da produção de distrofina, importante no reforço mecânico da membrana sarcolemal das fibras musculares. Degeneração progressiva e irreversível é inicialmente de predomínio proximal da musculatura esquelética. Qualidade de Vida (QV) inclui subjetividade, multidimensionalidade, presença de aspectos negativos e positivos diante da percepção e da expectativa individual de vida, com influência cultural. JUSTIFICATIVA: Não há estudos específicos para a DMD que apontem a análise da QV em diferentes faixas etárias evolutivas, sob diversos domínios, ou mesmo que indiquem precisamente qual domínio e idade específica devem receber maior atenção preventiva e terapêutica. OBJETIVOS: A partir da aplicação de três questionários de QV em diferentes fases evolutivas e domínios, objetivamos comprovar a praticidade e eficácia do Questionário Life Satisfaction Index for Adolescents - LSI-A, comparando-o aos Autoquestionnaire Qualitè de Vie Enfant Imagè - AUQUEI e Medical Outcomes Studies 36-item Short-Form - MOS SF-36. Essa pesquisa também possibilitará uma análise que subsidie o acompanhamento clínico, a prevenção das complicações, quando possível, e a corticoterapia paliativa. METODOLOGIA: Noventa e cinco pacientes foram divididos em 04 grupos, de acordo com as idades proporcionais às etapas da evolução da doença, por ocasião do início do acompanhamento: entre 5 e 7 anos (grupo A); entre 8 e 10 anos (grupo B); entre 11 e 13 anos (grupo C); 13 a 17anos de idade (grupo D). Os questionários (AUQEI, SF-36 e LSI-A) foram aplicados aos três, seis meses, 9 meses e até completar um seguimento de 12 meses da medicação. Na aplicação dos questionários, avaliamos a concordância entre dois observadores (confiabilidade inter-observador) e a concordância entre as observações feitas por um mesmo observador, em diferentes ocasiões. RESULTADOS: Os dados recolhidos e estaticamente representados denotam o grau de significância, ou qualidade de vida, do paciente nos diversos setores de sua vida, abordados em cada domínio dos questionários adotados. A média de significância geral foi detectada, mediante o questionário AUQEI, principalmente no grupo B. No SF-36, dos 8 domínios, somente 4 acusaram significância, e preponderantemente nos grupos B e C. Já no LSI-A, a significância foi geral à todos os grupos (A, B, C e D). CONCLUSÃO: Logo, pudemos inferir que os aspectos clínicos quando considerados em maior peso, como no SF- 36 e no AUQEI, mostram valores muito negativos na QV. O contrário foi constatado no questionário LSI-A, o qual atende, em sua natureza, às exigências de uma avaliação de qualidade de vida para crianças com Distrofia Muscular de Duchenne, pois abarca uma diversidade maior de circunstâncias na vida do paciente. / INTRODUTION: The Duchenne muscular dystrophy (DMD) is a genetic disease, which leads in changes in production of the dystrophin, important on reinforce mechanic of the sarcolemal membrane of the muscles fibers. The degeneration progress and irreversible is initially of the predominance proximal of the skeletal muscle. Quality of Life (QoL) includes subjectivity, multidimensionality, and presence of negatives and positives aspects in front of perception and expectation personal of life with cultural influence. JUSTIFICATION: there are no specific studies to the DMD, which had indicate a analysis of QoL into different ages and level evolutionary, under various domains, or studies which indicate which domain and specific age need to receive more preventive and therapeutic attention. OBJECTIVES: After of the application of three QoL questionnaires about different evolutionary phases and domains, we pretend to comprove the practicality and effectiveness of the Life Satisfaction Index for Adolescents LSI-A questionnaire, it comparing it to the Autoquestionnaire Qualitè de Vie Enfant Imagè - AUQUEI e Medical Outcomes Studies 36-item Short-Form - MOS SF-36. It is possible in this study also a analysis to the clinic support, prevention of the complications, and if enable, the palliative steroids therapy. METHODS: Ninety-five patients were divided into 04 groups, of according with the ages, proportional to the stages of the development of the disease at the initially of the accompaniment: between 5 and 7 years (group A), 8 and 10 years (group B), 11 and 13 years (group C); since 13 years of age (group D). The questionnaires (AUQEI, SF-36 and LSI-A) were applied to the three, six, nine months, and until they had completed a following of 12 months of medication. In the application of the questionnaires, we evaluated the concordance between two observers (inter-observer reliability), agreement between the observations made by the same observer on occasions different. RESULTS: The data collected and represented statically denotes the degree of significance or patients Qol in different areas of your life, all they mentioned in the domains of the QoL questionnaires used. The average of the overall significance was detected by the questionnaire AUQEI, especially in group B. In the SF-36, of the eight domains, only four had significance, and mainly in groups B and C. Already in the LSI-A, the significance was general for all groups (A, B, C and D). CONCLUSION: We infer that the clinical aspects, when considered in greater weight like on the SF-36 and AUQEI show negative values very in QOL. The opposite was noted in the LSI-A questionnaire, which serves in its nature to the requirements of an evaluating of QoL for children with DMD, because it covers the great diversify of circumstances in the patient life.