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1.
Reprod Sci ; 2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-33174187

RESUMO

The present study evaluated the effects of protocatechuic acid (PCA) after cisplatin-induced ovarian toxicity in mice and if PTEN and FOXO3a proteins are involved in PCA action. The mice were divided into five experimental groups (five animals per group) and treated once a day for 3 days as follows: (1) the control group was pretreated with oral administration (o.p.) of saline solution, followed by an intraperitoneal (i.p.) injection of saline solution. The other groups were pretreated (o.p.) with (2) saline solution (cisplatin group), (3) N-acetylcysteine (150 mg/kg of body weight), or with (4) 20 or (5) 50 mg/kg body weight of PCA, followed by 5 mg/kg body weight (i.p.) of cisplatin. Next, the ovaries were destined to histological (morphology and activation), immunohistochemical (PCNA and cleaved caspase-3 expression), and fluorescence (reactive oxygen species [ROS], glutathione [GSH], and active mitochondria levels) analyses. Moreover, the immunoreactivity for p-PTEN and p-FOXO3a was evaluated to investigate a potential mechanism by which PCA could prevent the cisplatin-induced ovarian damage. Pretreatment with N-acetylcysteine or 20 mg/kg PCA before cisplatin preserved the percentage of normal follicles and cell proliferation as observed in the control, reduced apoptosis and ROS levels, and showed higher active mitochondria and GSH levels than the cisplatin treatment (P < 0.05). Moreover, pretreatment with 20 mg/kg PCA decreased cisplatin-induced p-PTEN and increased (P < 0.05) nuclear export of p-FOXO3a. In conclusion, PCA at 20 mg/kg reduced apoptosis, maintained cell proliferation and mitochondrial function, reduced ROS production, and increased GSH expression likely through the involvement of PTEN and FOXO3a proteins.

2.
Cryobiology ; 91: 77-83, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31639331

RESUMO

This study aimed to evaluate different vitrification methods using distinct cryoprotectants (CPAs) for the preservation of collared peccary ovarian preantral follicles (PFs). Ovarian pairs from six females were fragmented and three fragments (fresh control group) were immediately evaluated for morphology, viability, cell proliferation capacity (assessed by quantifying the number of argyrophilic nucleolus organizer regions - NORs), and apoptosis (by the identification of activated caspase-3 expression). The remaining 18 fragments were vitrified using the solid surface vitrification (SSV) method or the ovarian tissue cryosystem (OTC) with 3 M ethylene glycol (EG), 3 M dimethylsulfoxide (DMSO), or a combination of the two (1.5 M EG/1.5 M DMSO). After two weeks, samples were rewarmed and evaluated as described previously. The OTC with any of the CPAs provided a similar conservation of morphologically normal PFs as the fresh control group (75.6 ±â€¯8.6%); however, the SSV was only efficient with DMSO alone (63.9 ±â€¯7.6%). Regarding the viability or cell proliferation, all tested groups provided post rewarming values similar to those observed for the fresh control group, 84.0 ±â€¯2.9% viable cells with 2.0 ±â€¯0.2 NORs. Related to apoptosis analysis, only the OTC with EG (46.7%) and the SSV method with EG (43.4%) or the combination of EG and DMSO (33.4%) provided similar values to those found for the fresh control group (36.7%). Our findings indicate the utilization of a closed system, the OTC, with 3 M EG as the CPA for the vitrification of collared peccary ovarian tissue.


Assuntos
Artiodáctilos/fisiologia , Criopreservação/veterinária , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Etilenoglicol/farmacologia , Folículo Ovariano/fisiologia , Animais , Apoptose/efeitos dos fármacos , Contagem de Células , Proliferação de Células/efeitos dos fármacos , Criopreservação/métodos , Feminino , Folículo Ovariano/citologia , Vitrificação
3.
Anim Reprod ; 16(4): 819-828, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-32368259

RESUMO

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.

4.
Rev Col Bras Cir ; 40(2): 130-6, 2013.
Artigo em Inglês, Português | MEDLINE | ID: mdl-23752640

RESUMO

OBJECTIVE: To develop an experimental model of stable saccular aneurysm in carotid of pigs using the internal jugular vein. METHODS: In 12 healthy pigs, weighing between 25 and 5 0kg, five males and seven females, we made a right common carotid artery aneurysm. After elliptical arteriotomy, we carried out a terminolateral anastomosis with the distal stump of the internal jugular vein. Aneurysm volume was calculated so that the value did not exceed 27 times the area of the arteriotomy. After six days angiography and microscopic examination were performed to assess patency of the aneurysm and the presence of total or partial thrombosis. RESULTS: There was a significant weight gain of pigs in the time interval between the manufacture of the aneurysm and angiography (p = 0.04). Aneurysmal patency was observed in ten pigs (83%). Operative wound infections occurred in two animals (16.6%), both with early onset, three days after the making of the aneurysm. Histological analysis showed aneurysm thrombus partially occluding the light in nine pigs (75%). In these animals, it was observed that on average 9% of the aneurysmal diameter was filled with thrombi. CONCLUSION: It was possible to develop a stable experimental model of saccular aneurysms in pig carotid artery by use of the internal jugular vein.


Assuntos
Aneurisma , Doenças das Artérias Carótidas , Modelos Animais de Doenças , Veias Jugulares , Animais , Feminino , Masculino , Suínos
5.
Rev. Col. Bras. Cir ; 40(2): 130-136, mar.-abr. 2013. ilus, tab
Artigo em Português | LILACS | ID: lil-676367

RESUMO

OBJETIVO: Desenvolver um modelo experimental estável de aneurisma sacular em carótida de suínos utilizando veia jugular interna. MÉTODOS: Em 12 suínos sadios, com peso variando entre 25 e 50kg, cinco machos e sete fêmeas, foi confeccionado aneurisma na artéria carótida comum direita. Após arteriotomia elíptica, foi realizada anastomose terminolateral com coto distal de veia jugular interna. O volume do aneurisma era calculado de maneira que o valor não excedesse em 27 vezes o valor da área da arteriotomia. Após seis dias, era realizada angiografia e análise microscópica do aneurisma para avaliar perviedade e trombose parcial ou total. RESULTADOS: Houve ganho de peso significante dos suínos no intervalo de tempo entre a confecção do aneurisma e a angiografia (p = 0,04). Foi observada perviedade aneurismática em dez suínos (83%). Ocorreram infecções de feridas operatórias em dois animais (16,6%), ambas com início de aparecimento em três dias após a confecção do aneurisma. Análise histológica dos aneurismas mostrou trombos ocluindo parcialmente a luz em nove suínos (75%). Nesses animais, observou-se que, em média, 9% da luz aneurismática estava preenchida por trombos. CONCLUSÃO: Pôde ser desenvolvido um modelo experimental estável de aneurisma sacular em carótida de suínos utilizando veia jugular interna.


OBJECTIVE: To develop an experimental model of stable saccular aneurysm in carotid of pigs using the internal jugular vein. METHODS: In 12 healthy pigs, weighing between 25 and 50kg, five males and seven females, we made a right common carotid artery aneurysm. After elliptical arteriotomy, we carried out a terminolateral anastomosis with the distal stump of the internal jugular vein. Aneurysm volume was calculated so that the value did not exceed 27 times the area of the arteriotomy. After six days angiography and microscopic examination were performed to assess patency of the aneurysm and the presence of total or partial thrombosis. RESULTS: There was a significant weight gain of pigs in the time interval between the manufacture of the aneurysm and angiography (p = 0.04). Aneurysmal patency was observed in ten pigs (83%). Operative wound infections occurred in two animals (16.6%), both with early onset, three days after the making of the aneurysm. Histological analysis showed aneurysm thrombus partially occluding the light in nine pigs (75%). In these animals, it was observed that on average 9% of the aneurysmal diameter was filled with thrombi. CONCLUSION: It was possible to develop a stable experimental model of saccular aneurysms in pig carotid artery by use of the internal jugular vein.


Assuntos
Animais , Feminino , Masculino , Aneurisma , Doenças das Artérias Carótidas , Modelos Animais de Doenças , Veias Jugulares , Suínos
6.
Reprod Fertil Dev ; 24(7): 905-15, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22935151

RESUMO

Several growth factors have been identified as local regulators of follicle development and ovulation. Fibroblast growth factor (FGF) family members are potent mitogens and are involved in cell differentiation, cell migration and angiogenesis in many tissues and organs. In addition to FGF-2, which is the most-studied FGF, other important members are FGF-1, -5, -7, -8, -9 and -10. A number of studies have indicated that FGFs play important roles in regulating the initiation of primordial follicle growth, oocyte and follicle survival, granulosa and theca cell proliferation and differentiation, corpus luteum formation, steroidogenesis and angiogenesis. The purpose of this review is to highlight the importance of the FGFs on mammalian female reproduction, providing a better understanding of the roles of this family in ovarian physiology and female fertility.


Assuntos
Fatores de Crescimento de Fibroblastos/metabolismo , Folículo Ovariano/metabolismo , Ovulação , Animais , Sobrevivência Celular , Feminino , Fertilidade , Humanos , Ligantes , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Transdução de Sinais
7.
Braz. j. vet. res. anim. sci ; 46(5): 378-386, 2009. ilus, tab, graf
Artigo em Inglês | LILACS | ID: lil-538430

RESUMO

The aim of this study was to evaluate the effects of pituitary (pFSH) or recombinant (rFSH) FSH on the survival and growth of caprine preantral follicles. Caprine ovarian tissues were in vitro cultured for one or seven days in Minimum Essential Medium (MEM) alone or containing 10, 50, 100 and 1000 ng/ml of pFSH or rFSH. Control tissues (non-cultured) and those cultured were processed for histological and ultrastructural studies. In addition, follicular and oocyte diameter were analysed. After seven days of culture, only 50ng/ml of rFSH maintained the percentage of normal follicles similar to control. Moreover, 10 ng/ml of pFSH and all the concentrations of rFSH promoted primordial follicles activation. In addition, the presence of 50 ng/ml of rFSH promoted the highest follicular diameter at day seven of culture. In conclusion, 50 ng/ml of rFSH maintained the ultrastructural integrity of caprine preantral follicles, promoted primordial follicles activation and further growth of cultured follicles.


O objetivo deste estudo foi avaliar os efeitos do FSH pituitário (pFSH)ou recombinante (rFSH) sobre a sobrevivência e o crescimento de folículos pré-antrais caprinos. O tecido ovariano foi cultivado in vitro por um ou sete dias em Meio Essencial Mínimo (MEM) sozinho, ou contendo 10, 50, 100 e 1000 ng/ml de pFSH ou rFSH. O grupo controle (não cultivado) e aqueles cultivados foram processados para análises histológica e ultra-estrutural. Além disso, os diâmetros folicular e oocitário foram avaliados. Após sete dias de cultivo, apenas 50 ng/ml de rFSH manteve o percentual de folículos normais semelhante ao controle. Além disso, 10 ng/ml de pFSH e todas as concentrações de rFSH promoveram ativação de folículos primordiais. A presença de50 ng/ml de rFSH promoveu o maior diâmetro folicular após sete dias de cultivo. Em conclusão, 50 ng/ml de rFSH manteve a integridadede folículos pré-antrais caprinos e promoveu a ativação e o crescimento dos folículos cultivados.


Assuntos
Animais , Hormônio Foliculoestimulante , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/ultraestrutura , Cabras , Meios de Cultura , Técnicas de Cultura de Órgãos
8.
Anim Reprod Sci ; 108(3-4): 309-18, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17945440

RESUMO

Bovine preantral follicles within ovarian fragments were exposed and cryopreserved in absence or presence of 1.5M glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO), undergoing a previous cooling at 20 degrees C for 1h (protocol 1) or at 4 degrees C for 24h (protocol 2) in 0.9% saline solution. At the end of each treatment, preantral follicles were classified as non-viable/viable when they were stained/not stained with trypan blue, respectively. To confirm viability staining, ultra-structure of the follicles was evaluated by transmission electronic microscopy (TEM). Data were compared by Chi-square test (P<0.05). The storage of the ovaries at 20 degrees C for 1h (78%) and 4 degrees C for 24h (80%) did not reduce significantly the percentage of viable preantral follicles when compared to the control (75%). Similar results were obtained when ovarian fragments, respectively, for protocols 1 and 2, were exposed to MEM (78 and 77%), 1.5M EG (78 and 71%), as well as frozen in 1.5M EG (74 and 77%). Percentages of viable follicles in control were similar to those observed after exposure (75%) and freezing (76%) in presence of 1.5M DMSO only when protocol 1 was used. The increase of the concentration from 1.5 to 3.0M, for all cryoprotectants, reduced significantly the percentage of viable preantral follicles after freezing. Ultra-structural analysis has confirmed trypan blue results, showing that not only basement membrane, but also organelles, were intact in viable preantral follicles. In conclusion, ovarian tissue cooling at 4 degrees C for 24h before cryopreservation (protocol 2) does not affect the viability of bovine preantral follicles when 1.5M EG is present in the cryopreservation medium.


Assuntos
Bovinos/fisiologia , Criopreservação/veterinária , Crioprotetores/farmacologia , Folículo Ovariano/fisiologia , Animais , Distribuição de Qui-Quadrado , Criopreservação/métodos , Dimetil Sulfóxido/farmacologia , Etilenoglicol/farmacologia , Feminino , Glicerol/farmacologia , Microscopia Eletrônica de Transmissão/veterinária , Folículo Ovariano/ultraestrutura , Propilenoglicóis/farmacologia , Azul Tripano/química
9.
Braz. j. vet. res. anim. sci ; 43(2): 250-255, 2006. graf
Artigo em Português | LILACS | ID: lil-454662

RESUMO

O objetivo deste estudo foi avaliar foliculos pré-antrais (FOPA) ovinos isolados após sua exposição e criopreservação utilizando glicerol (GLI), etilenoglicol (EG), propanodiol (PROH) ou dimetilsulfóxido (DMSO) a 1,5 e 3,0 M. Cada par ovariano de 5 ovelhas sem raça definida foi coletado em abatedouro local e submetido ao isolamento folicular. Da suspensão obtida, uma aliquota foi imediatamente destinada à análise da viabilidade folicular com o auxílio do corante vital azul de trypan. O restante da suspensão foi dividida em 16 aliquotas de 0,9 mL, suspensas (v/v) em MEM+ com EG, DMSO, GLI ou PROH a 1,5 ou 3,0 M, para teste de toxicidade e criopreservação. Após o término de cada tratamento, a viabilidade folicular foi analisada e os FOPA considerados viáveis se não corados ou não viáveis, quando corados. A análise dos dados mostrou que após o teste de toxicidade e criopreservação, em todos os crioprotetores e em ambas as concentrações, a percentagem de FOPA viáveis foi significativamente reduzida quando comparada ao controle. No teste de toxicidade, quando os crioprotetores foram comparados entre si nas mesmas concentrações, foram observadas percentagens signifIcativamente menores de FOPA viáveis no PROH 3,0 M (38,9%), apresentando-se, portanto, mais tóxico quando comparado aos demais crioprotetores. Após criopreservação, obteve-se percentagens significativamente maiores de foliculos pré-antrais viáveis quando o EG e o DMSO foram utilizados. Em conclusão, FOPA ovinos isolados podem ser criopreservados com sucesso utilizando-se D MSO e EG a 1,5 e 3,0 M.


The aim of this study was to evaluate isolated sheep preantral follicles (PF) after exposure and cryopreservation using glycerol (GLI), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO) at 1.5 and 3.0 M. Each ovarian pair from 5 mixed breed adult sheeps was obtained at a local slaughterhouse and submited to follicular isolation. From the obtained suspension, one aliquot was immediately analysed with trypan blue. The remaining suspension was divided in 16 aliquots of 0.9 mL, suspended in (v /v) in MEM+with EG, DMSO, GLI or PROH at 1.5 or 3.0 M to the toxicity test and cryopreservation. After the end of each treatment, the follicular viability was analysed and the PF were classified as viable if not dyed or not viable if dyed with trypan blue. The analysis of the results showed that after the toxicity test and cryopreservation, using all cryoprotectants and at both concentrations, the percentage of viable PF was significandy reduced when compared to the control. At the toxicity test, when the cryoprotectants were compared at the same concentrations, the lowest percentage of viable preantral follicles was obtained when 3.0 M PRO H (38,9%) was used, being, more toxic when compared to the others cryoprotectants. After cryopreservation, significantly higher percentual of viable PF was observed when the EG and DMSO were used. In conclusion, sheep PF can be cryopreserved successfully using DMSO and EG at 1.5 and 3.0 M.


Assuntos
Criopreservação/veterinária , Folículo Ovariano/anatomia & histologia , Ovário/anatomia & histologia , Ovário/metabolismo , Óvulo/crescimento & desenvolvimento , Ovinos , Testes de Toxicidade/veterinária
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