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Effect of IL-10 and TNF-α on the competence and cryosurvival of in vitro produced Bos indicus embryos

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Autor(es):
Silva, Mara Viana ; Valente, Roniele Santana ; Annes, Kelly ; Marsico, Thamiris Vieira ; Oliveira, Andressa Minozzo ; Maiollo, Bruna Aparecida Prado ; Lopes, Nelyane Jose ; Tannura, Juliana Hayashi ; Sudano, Mateus Jose
Número total de Autores: 9
Tipo de documento: Artigo Científico
Fonte: Theriogenology; v. 215, p. 7-pg., 2023-12-09.
Resumo

In vitro-produced embryos are constantly exposed to stressful conditions that can lead to the activation of the apoptotic pathway. The nuclear Kappa B factor (NF-kappa B) is an inflammatory mediator that induces the expression of tumor necrosis factor (TNF-alpha), a pro-inflammatory cytokine, while interleukin-10 (IL-10), an anti-inflammatory cytokine, inhibits NF-kappa B activity. This study aimed to investigate the effects of IL-10 and TNF-alpha on the competence and cryosurvival of in vitro-produced bovine embryos. Embryos were produced in vitro using standard protocols, and Grade I blastocysts were vitrified using the Cryotop method. Non-vitrified and vitrified blastocysts were subjected to the TUNEL assay. In Experiment I, on day 6.5 (156 h post-insemination), the embryos were treated with PBS (control), 50 ng/mL of IL-10, or a combination of 25 ng/mL of TNF-alpha and 50 ng/mL of IL-10. Embryonic development and apoptotic rates were monitored. In Experiment II, the same groups were set up, with the addition of a group treated with 25 ng/mL of TNF-alpha alone. Grade I blastocysts were vitrified 5 h after treatment, and cryosurvival was monitored at until 48 h post-warming. The apoptosis rate and total cell number were investigated in the vitrified-hatched blastocysts. IL-10 alone did not affect developmental competence or cryosurvival (P > 0.05). The IL-10-treated embryos, when exposed in combination with TNF-alpha, presented a detrimental effect (P < 0.05) in the embryonic development of non-vitrified embryos. However, vitrified blastocysts had no negative effect (P > 0.05). The TNF-alpha treatment reduced (P < 0.05) the re-expansion rate at 6 h post-warming and increased (P < 0.05) the apoptosis rate in vitrified hatched blastocysts, whereas no effect (P > 0.05) of the treatments was detected in the hatching rate and total cell number post-warming. In conclusion, TNF-alpha has a detrimental effect on embryonic developmental competence and cryosurvival by compromising the development of non-vitrified embryos and apoptotic-related events of vitrified blastocysts, whereas IL-10, when in combination with TNF-alpha, appears to attenuate the detrimental effects of TNF-alpha. (AU)

Processo FAPESP: 21/01446-9 - EMU concedido no processo 18/21496-8: Microscópio Invertido de Fluorescência e Contraste de interferência Diferencial (DIC)
Beneficiário:Mateus José Sudano
Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
Processo FAPESP: 19/17796-9 - Biopsia simultânea e perfil lipídico da massa celular interna e trofectoderma embrionário
Beneficiário:Thamiris Vieira Marsico
Modalidade de apoio: Bolsas no Brasil - Doutorado
Processo FAPESP: 18/21496-8 - Caminhos para prenhez embrionária
Beneficiário:Mateus José Sudano
Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
Processo FAPESP: 20/08826-9 - Investigação do sistema de membranas celulares e sua relação com a criotolerância e viabilidade de embriões bovinos produzidos in vitro
Beneficiário:Kelly Annes
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado