Navegando por Palavras-chave "líquido folicular"
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- ItemSomente MetadadadosAnálise dos perfis proteômico e lipidômico de líquido folicular de pacientes com endometriose e endometrioma submetidas ao tratamento de fertilização in vitro(Universidade Federal de São Paulo (UNIFESP), 2013-05-14) Regiani, Thais [UNIFESP]; Lo Turco, Edson Guimaraes [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Objetivo: Determinar o perfil proteico e lipídico do líquido folicular de pacientes com endometriose e endometrioma submetidos ao tratamento de fertilização in vitro determinando assim possíveis candidatos proteicos e lipídicos para a endometriose. Métodos: Amostras do líquido folicular de 10 mulheres com endometriose grau III ou IV, diagnosticadas por videolaparoscopia, e seus respectivos endometriomas ovarianos, foram coletadas. O grupo controle foi composto por 10 mulheres que tiveram sucesso na gravidez após o tratamento de fertilização in vitro. Para proteômica as amostras foram agrupadas em pools, de acordo com a concentração de proteínas das mesmas e para essa análise utilizou-se a técnica de 2D nanoUPLC-ESI-MSE. O processamento dos espectros e a identificação das proteínas, foram realizados com auxílio do software ProteinLynx Global Server v.2.5 (PLGS). As redes biológicas foram produzidas no programa Cytoscape 2.8.2 com o plugin ClueGO. Para a análise de lipidômica, realizouse a extração de lipídeos, utilizando o protocolo de Bligh e Dyer. A fase apolar foi analisada por um espectrômetro do tipo ESI-QTOF, em modo positivo e a análise estrutural dos possíveis biomarcadores foram realizadas por análise de MS/MS. Resultados: Um total de 535 proteínas foram identificadas e quantificadas. Entre as proteínas exclusivas e/ou diferencialmente expressas, 33 pertencem ao grupo Controle, 41 ao grupo Endometriose (OSE) e 212 ao grupo Endometrioma (OMA). Pela análise de enriquecimento funcional pôde-se observar no grupo OSE a presença de funções de regulação relacionadas à secreção de citocinas, processos de coagulação, transporte, ligação e metabolismo de esteroides. No grupo OMA foi possível encontrar proteínas relacionadas ao aumento dos danos de DNA, ao processo inflamatório, estresse oxidativo, apoptose e produção exacerbada de espécies reativas ao oxigênio. Na análise de lipidômica, no grupo controle foram encontradas fosfatidilglicerol fosfato, fosfatidilcolina, fosfatidilserina e fosfatidilnositol bifosfato. Nos grupos de estudo as subclasses lipídicas hiper-representas foram: fosfatidilcolina e esfingomielina. Conclusões: O perfil de proteínas e lipídeos é diferente entre mulheres com e sem endometriose. As alterações de ambos os perfis do líquido folicular mostraram que essa doença pode afetar a fisiologia folicular.
- ItemSomente MetadadadosA influência do índice de massa corporal na fisiologia do folículo ovariano em ciclos de reprodução assistida(Universidade Federal de São Paulo (UNIFESP), 2013-04-03) Porciuncula, Patricia Marafon [UNIFESP]; Bertolla, Ricardo Pimenta Bertolla [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Objective: To observe the influence of Body Mass Index (BMI) on the follicular metabolism of women submitted to controlled ovarian hyperstimulation. Methods: A prospective study was carried out including 129 follicular fluid samples from women submitted to controlled ovarian hyperstimulation and classified, according to BMI, in three groups according to the World Health Organization: eutrophic (20 to <25kg/m2), overweight (25 to <30kg/m2), and obese (>30kg/m2). Follicular fluid samples were obtained by ultrasound-guided transvaginal ovarian aspiration following controlled ovarian hyperstimulation. Immediately after collection, the follicular fluid samples were centrifuged to remove cellular debris and frozen until analysis. For quantification of the steroid hormones estradiol, progesterone, testosterone, androstenedione, and cortisol, tandem mass spectrometry was utilized (ID-LC-MS/MS). For proteomic analysis, three pools were formed (one for each group, normalized to total protein in each sample), and proteins were quantified and identified utilizing the shotgun proteomics approach nanoUPLC-nanoESI-MSE, resulting in ?Identified? and ?Quantified and Identified? proteins. Proteins observed in only one replicate were excluded from the study. For steroid profiling, groups were compared using one-way ANOVA followed by a Least Significant Differences posthoc test, or using a Kruskal-Wallis followed by a Mann-Whitney test. For the proteomics experiment a Kruskal-Wallis test was used, followed by a Mann-Whitney test. An alpha of 5% was adopted in the study. Protein lists were utilized for functional enrichment analysis. Results: Regarding the steroid profile, differences were observed between the BMI groups. The obesity group presented lower progesterone levels and higher androstenedione levels when compared to the other two groups. Also, estradiol, testosterone, and cortisol were higher in both overweight and obese groups when compared to eutrophic controls. Regarding proteomics, 284 proteins were identified, of which 152 were quantified. Major enriched functions were related to immune response, inflammation and coagulation. Moreover, seven proteins were hyperexpressed only in the obese patients. Finally, 20 proteins were exclusively expressed in the obesity group, 10 in the overweight group, and 19 in the eutrophic group. Conclusion: Follicular fluid metabolism is altered due to obesity in patients submitted to controlled ovarian hyperstimulation. The follicular fluid of overweight and obese patients presents altered steroid profiles, with increased estradiol, testosterone, and cortisol, and obese patients present increased androstenedione and decreased progesterone. Also, overweightness and obesity alter the follicular fluid protein profile of patients submitted to controlled ovarian hyperstimulation, with enriched immune and inflammatory response functions.
- ItemSomente MetadadadosInfluência dos fatores femininos de infertilidade no estresse oxidativo do líquido folicular de pacientes submetidas a estímulo ovariano controlado(Universidade Federal de São Paulo (UNIFESP), 2014-04-15) Lima, Camila Bruna de [UNIFESP]; Lo Turco, Edson Guimaraes [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Purpose: The present study aimed to evidence the oxidative status of follicular fluid from women submitted to in vitro fertilization treatments in order to establish a relationship between oxidative stress (OS) and the pathophysiology of different diseases like Polycystic Ovary Syndrome (PCOS) and Endometriosis. This study also aimed to correlate the OS with factors that may influence the success of assisted reproduction techniques such as the ovarian response to exogenous hormonal stimulation. Methods: This study included 183 patients referred to the Human Reproduction Service of Sao Paulo Hospital for in vitro fertilization treatments. They were separated according to the fertility factor to form the following groups: Controls (n=49), Endometriosis (n=48), Poor response (n=26), PCOS (n=15) e Hyper response (n=46). All patients were submitted to controlled ovarian hyper stimulation protocols and ovarian punction for follicular fluid collection. Results: The levels of oxidative stress were significantly higher in the groups Endometriosis (p=9,57x10-13), PCOS (p=0,00015) and Hyper response (p=7,04x10-7) when compared to Control group. In endometriosis group, it is possible to observe a positive correlation between FSH and LH serum levels (r=0,315 e p=0,031), and also between FSH and age (r=0,306 e p=0,035). It was not possible to find significant differences in OS levels on the comparison between Poor response and Control groups. (p=0,808). The moving average analysis showed a trend for increasing oxidative status with increasing age in patients who had hyper response to ovarian stimulation. It also showed a reduction in OS levels with increasing age in Endometriosis group. Pregnancy rates did not present statistically relevant differences among groups (p=0,381). Conclusions: In Endometriosis, PCOS and Hyper response groups, higher amounts of end products of lipid peroxidation were detected in follicular fluid. This highlights an elevated oxidative status in these patients. Moreover, the age seems to influence the levels of oxidative stress in Endometriosis and Hyper response groups. However, it was not possible to correlate oxidative stress with poor response or pregnancy rates.