Protective effect of kaempferol against cisplatin-induced acute ovarian damage in a mouse model / [Efeito protetor do kaempferol contra o dano ovariano agudo induzido pela cisplatina em um modelo murino]

The flavonoid kaempferol has attracted research attention as a potential adjuvant during chemotherapy. This study aimed to evaluate the protective effects of kaempferol against ovarian damage in cisplatin-treated mice. Two groups of mice received saline solution (intraperitoneal injection [i.p.]; control) or a single dose of cisplatin (5 mg/kg body weight, i.p.). Moreover, two other mice groups were pretreated with kaempferol (1 or 10 mg/kg body weight, i.p.) 30 min before of the cisplatin administration. Thereafter, their ovaries were harvested and subjected to histological (follicular morphology and activation) and fluorescence (reactive oxygen species [ROS] production, glutathione [GSH] concentration, and mitochondrial activity) analyses. Compared with cisplatin treatment alone, pretreatment with 1 mg/kg kaempferol maintained normal follicular morphology, reduced ROS production and mitochondrial damage, and enhanced GSH concentration. However, pretreatment with 10 mg/kg kaempferol did not prevent cisplatin-induced damage. The rate of primordial follicle activation was greater in mice pretreated with 1 mg/kg kaempferol than in the other treatment groups. In conclusion, pretreatment with 1 mg/kg kaempferol prevents cisplatin-induced ovarian damage and stimulates primordial follicle activation in mice.

O flavonoide kaempferol tem atraído a atenção como um potencial adjuvante durante a quimioterapia. O presente estudo objetivou avaliar os efeitos do kaempferol contra os danos ovarianos em camundongos tratados com cisplatina. Fêmeas de camundongos receberam solução salina (injeção intraperitoneal [ip]; controle) ou uma dose única de cisplatina (5 mg/kg, ip) ou foram pré-tratadas com kaempferol (1 ou 10 mg/kg, ip) 30 min antes da administração de cisplatina. Os ovários foram recuperados e destinados para as análises histológicas (morfologia e ativação folicular) e de fluorescência (produção de espécies reativas de oxigênio [ERO], concentração de glutationa [GSH] e atividade mitocondrial). Em comparação ao tratamento apenas com cisplatina, o pré-tratamento com 1 mg/kg de kaempferol manteve a morfologia folicular normal, reduziu a produção de ERO, bem como os danos mitocondriais, e aumentou a concentração de GSH. Entretanto, o pré-tratamento com 10 mg/kg de kaempferol não preveniu os danos induzidos pela cisplatina. A taxa de ativação do folículo primordial foi maior em camundongos pré-tratados com 1 mg/kg de kaempferol do que nos outros grupos experimentais. Em conclusão, o pré-tratamento com 1 mg/kg de kaempferol previne o dano ovariano induzido pela cisplatina e estimula a ativação do folículo primordial em camundongos.

Influence of the ovarian fragmentation before storage at 4o C on the apoptosis rates and in vitro development of ovine preantral follicles

The aim of this study was to evaluate the effect of ovarian tissue fragmentation before preservation at 4o C in MEM on the morphology, apoptosis, and growth of ovine preantral follicles. After collection, the ovaries were divided into two halves, being one divided into two fragments (1/4 of the ovary). One fragment was subdivided into two fragments (1/8), being one fixed for histology (fresh control). The remaining whole ovary, 1/2, 1/4 and 1/8 of the ovary were preserved in MEM at 4°C for 6, 12 or 24 h. The tissue was further destined to histology. In vitro culture and TUNEL technique were performed in treatments that showed the best results of follicular survival after preservation. Storage of 1/8 of the ovary increased the normal follicles compared with half or whole ovary. After preservation in 1/8 of the ovary and culture for 7 days, the percentage of apoptotic cells was similar to the fresh control and non-cultured fragments. The percentage of growing follicles increased after preservation of 1/8 of the ovary compared with 1/4. In conclusion, ovine preantral follicles can be preserved in fragments of 1/8 of the ovary in MEM at 4°C for 24 h, without affecting apoptosis and their ability to grow in vitro.

Influence of the ovarian fragmentation before storage at 4o C on the apoptosis rates and in vitro development of ovine preantral follicles

The aim of this study was to evaluate the effect of ovarian tissue fragmentation before preservation at 4o C in MEM on the morphology, apoptosis, and growth of ovine preantral follicles. After collection, the ovaries were divided into two halves, being one divided into two fragments (1/4 of the ovary). One fragment was subdivided into two fragments (1/8), being one fixed for histology (fresh control). The remaining whole ovary, 1/2, 1/4 and 1/8 of the ovary were preserved in MEM at 4°C for 6, 12 or 24 h. The tissue was further destined to histology. In vitro culture and TUNEL technique were performed in treatments that showed the best results of follicular survival after preservation. Storage of 1/8 of the ovary increased the normal follicles compared with half or whole ovary. After preservation in 1/8 of the ovary and culture for 7 days, the percentage of apoptotic cells was similar to the fresh control and non-cultured fragments. The percentage of growing follicles increased after preservation of 1/8 of the ovary compared with 1/4. In conclusion, ovine preantral follicles can be preserved in fragments of 1/8 of the ovary in MEM at 4°C for 24 h, without affecting apoptosis and their ability to grow in vitro.(AU)

Effect of ovarian tissue transportation in Amburana cearensis extract on the morphology and apoptosis of goat preantral follicles

The aim of the present study was to evaluate the effect of Amburana cearensis extract during caprine ovarian tissue transportation on the survival of preantral follicles in vitro. HPLC was used to determine the fingerprint chromatogram of the ethanolic extract. Five goat ovarian pairs were divided into fragments. One fragment was fixed for histology and TUNEL Analysis (fresh control). The other fragments were placed in MEM or A. cearensis extract (0.1; 0.2 or 0.4 mg/ml) and stored at 4oC for 6, 12 or 24 h. Preserved Fragments were also fixed for histology and TUNEL analysis. The presence of phenolic compounds (protocatechuic acid, epicatechin, p-coumaric acid, gallic acid and kaempferol) in the extract was confirmed using HPLC. Thepercentage of normal follicles preserved in 0.2 mg/ml A. cearensis for 6 h was similar to that observed in the fresh control. Moreover, the percentage of normal follicles was higher after preservation in 0.2 mg/ml A. cearensisFor 6 h than the other A. cearensis treatments and similar to that found in MEM. There were no differences in the percentage of apoptotic cells between fresh control and those preserved for 6 h in MEM or 0.2 mg/ml A. cearensis. In conclusion, both 0.2 mg/ml A. cearensis or MEM can be used for the preservation of goat preantral follicles for up to 6 h. The use of A. cearenses is recommended due to the higher cost of MEM.

Effect of ovarian tissue transportation in Amburana cearensis extract on the morphology and apoptosis of goat preantral follicles

The aim of the present study was to evaluate the effect of Amburana cearensis extract during caprine ovarian tissue transportation on the survival of preantral follicles in vitro. HPLC was used to determine the fingerprint chromatogram of the ethanolic extract. Five goat ovarian pairs were divided into fragments. One fragment was fixed for histology and TUNEL Analysis (fresh control). The other fragments were placed in MEM or A. cearensis extract (0.1; 0.2 or 0.4 mg/ml) and stored at 4oC for 6, 12 or 24 h. Preserved Fragments were also fixed for histology and TUNEL analysis. The presence of phenolic compounds (protocatechuic acid, epicatechin, p-coumaric acid, gallic acid and kaempferol) in the extract was confirmed using HPLC. Thepercentage of normal follicles preserved in 0.2 mg/ml A. cearensis for 6 h was similar to that observed in the fresh control. Moreover, the percentage of normal follicles was higher after preservation in 0.2 mg/ml A. cearensisFor 6 h than the other A. cearensis treatments and similar to that found in MEM. There were no differences in the percentage of apoptotic cells between fresh control and those preserved for 6 h in MEM or 0.2 mg/ml A. cearensis. In conclusion, both 0.2 mg/ml A. cearensis or MEM can be used for the preservation of goat preantral follicles for up to 6 h. The use of A. cearenses is recommended due to the higher cost of MEM.(AU)