Ação do resveratrol sobre os parâmetros reprodutivos de ratos tratados com cisplatina, etoposide e bleomicina a partir da peripuberdade

• Ano de defesa: 2016
• Autor(a) principal: Neves, Flavia Macedo de Oliveira [UNIFESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Resveratrol; Testis; Kratos; Bep protocol; Sperm; Testículo; Ratos; Protocolo bep; Espermatozoides
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=3674986; http://repositorio.unifesp.br/handle/11600/47718

Protocols that include co-administration of bleomycin, etoposide and cisplatin (BEP protocol) have been effective against this disease; however, cause deleterious effects on fertility. Resveratrol can be considered as potential candidate to reverse this panorama due to its anti-cancer, anti-apoptotic, anti-inflammatory and antioxidant. This study aimed to determine the effect of the co-administration of resveratrol against the side effects on reproduction of rats, caused by the administration of BEP in a clinical dose adapted for peripubertal rats. A hundred and seventy two rats were distributed into four groups: Sham Control (SC); Resveratrol (R); Bleomicin, Etoposide and Cisplatin (BEP) and Resveratrol + Bleomicin, Etoposide and Cisplatin (R-BEP). At 36th days postpartum (dpp), the rats from the groups R (n=44) and R-BEP (n=42) were treated with resveratrol, by gavage, with a single daily dose of 300 mg/kg body weight. From the 41st dpp, the animals of RBEP group were submitted to the co-administration BEP intraperitoneally (i.p.), for three consecutive weeks using the following protocol: 3.5 mg/kg etoposide and 0.70 mg/kg of cisplatin, for 5 consecutive days, with 02 days off treatment every week and 0.35 mg/kg of bleomycin on the 2nd day of every week. Animals of the BEP group (n=42) were exclusively treated with bleomycin, etoposide and cisplatin via ip; those of the SC group received saline solution via i.p. and carboxymethyl-cellulose (by gavage), which is the vehicle for resveratrol (n=44). The euthanasia was performed at two ages: i) 60 dpp (Experiment 1) and ii) 113 dpp (Experiment 2). In both ages the following parameters were investigated: testicular and epididymal biometry, testicular morphometry and stereology, and sperm quantitative analysis. The number of apoptotic germ cells (numerical density of TUNEL-positive cells) and the histopathological analysis of the testes were also assessed in both ages. At 60 dpp we also determined: dosages of sex hormones (testosterone and estradiol) in the plasma (P) and in the intratesticular fluid (ITF); the testicular levels of malondyaldehyde and nitric oxide were carried out to access the oxidative stress and qualitative sperm analysis (mitochondrial activity; acrosome integrity; DNA integrity; morphology and motility - CASA/HTM-IVOS). At 113 dpp the total estimate of spermatogonia RB1-positive was investigated. Neither in 60 nor in 113 dpp animals there were significant differences when CS and R groups were compared, in any of the parameters analyzed. At 60 dpp the animals of BEP and RBEP group showed a reduction of the following parameters: body weight, epididymis and testis weights, diameter of seminiferous tubules, including the androgendependent tubules, seminiferous epithelium height and area, both P an ITF testosterone dosages and the number of sperm in the different portions of the epididymis. Accentuated hypotrophy of the seminiferous epithelium (germ cell depletion) also occurred in this age in both groups, BEP and R-BEP. When the groups BEP and R-BEP where compared, no significant alterations were observed, except for the seminiferous epithelium area (R-BEP > BEP). Still in the 60 dpp, there was an increase in the testicular lipid peroxidation in both BEP and R-BEP groups. In the 60 dpp, the sperm parameters such as DNA integrity and motility (vigor - VCL and progression - VAP, VSL) showed to be similarly altered in both R-BEP and BEP groups, when compared to SC and R groups. However, the dosages of ITF testosterone and some parameters that indicate sperm quality (morphology, mitochondrial activity and acrosome integrity) point out to a benefic action of resveratrol against the damage caused by the chemotherapy, since a significant improvement in the R-BEP group was observed compared to the BEP group. However, the R-BEP group showed significant alterations in all the parameters analyzed, when compared to the SC group. At 113 dpp, the animals of BEP and RBEP groups presented similar alterations in the reproductive parameters as observed at 60 dpp, plus a reduction in the estimated number of total RB1-positive spermatogonia. There was an increase in the occurrence of germ cell death through apoptosis in the groups BEP and R-BEP (60 and 113 dpp), but at 60 dpp it was reduced in the animals from the R-BEP group, compared to the BEP group. The model of study presented here provided a broad analysis of the BEP protocol effects with or without the co-administration of resveratrol on different types of germ cells; it especially provided evidence on the impact of the treatments (BEP and R-BEP) on the survival of these cells and on the quality of the gametes, since the euthanasia was performed at two different time points: i) one day (Experiment 1) and ii) 54 days (Experiment 2) after the end of the treatment (at 59dpp). The reduction in the occurrence of germ cell apoptosis, the partial preservation of the seminiferous epithelium area, the improvement in some sperm qualitative parameters and in the ITF testosterone dosage observed in the R-BEP group (60 dpp), compared to the BEP group, indicate a positive action produced by the treatment with resveratrol, in the co-administration with BEP. On the other hand, resveratrol did not provide sufficient protective action against the chemotherapy to restore the seminiferous epithelium after the completion of a full spermatogenic cycle (53 days, 113 dpp). Longer periods of recovery elapsed after treatment could raise the chances of clarifying the role of resveratrol on the late restoration of the seminiferous epithelium and on the recovery of sperm reserves.