Efeito do meio condicionado por células tronco mesenquimais na resposta inflamatória uterina pós-cobertura em éguas susceptíveis
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Data
2020-12-03
Autores
Brasileiro, Lucas Santiago Gomes
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A endometrite é a principal causa de subfertilidade na espécie equina. Essa enfermidade caracteriza-se pelo acúmulo de líquido no interior do útero, influxo de células polimorfonucleares para o lúmen uterino, exacerbada expressão de mediadores pró-inflamatórios e, consequentemente, baixas taxas de fertilidade. Nos mamíferos em geral, logo após o contato do sêmen com o endométrio, ocorre uma resposta inflamatória a fim de eliminar espermatozoides mortos, debris celulares, plasma seminal e micro-organismos que são carreados no momento da cópula/inseminação artificial (IA). As éguas podem ser caracterizadas como resistentes ou susceptíveis a endometrites; aquelas que são resistentes possuem a capacidade de modular a resposta inflamatória uterina até 48 horas após a cobertura/IA, enquanto as susceptíveis não tem essa habilidade. Os tratamentos mais comumente utilizados para a endometrite são as lavagens uterinas, administração de drogas ecbólicas, antibióticos e anti-inflamatórios. Com o avanço de técnicas de terapia celular com capacidade de modular a resposta inflamatória, diferentes tratamentos vêm sendo estudados com resultados promissores no tratamento da endometrite em éguas. Um modulador de resposta inflamatória que vem sendo estudado e se mostra como uma possibilidade terapêutica para o tratamento de éguas com Endometrite Persistente Pós-Cobertura (EPPC) é o meio condicionado de células tronco (MC). Dessa forma, o presente estudo teve por objetivo revisar os fatores relacionados a endometrite em éguas e o desenvolvimento de terapias celulares, em especial as células tronco e o meio condicionado (MC) de células tronco mesenquimais. Foram selecionadas éguas com histórico reprodutivo conhecido, classificadas como susceptíveis a EPPC. Eram realizados acompanhamentos diário por palpação retal e ultrassonografia até o momento da indução da ovulação. As éguas eram inseminadas em ciclos estrais consecutivos. Para o ciclo controle, as éguas não receberam nenhum tratamento pré IA. No ciclo tratado, 30 mL de MC foi infundido no útero das éguas 24 horas antes da IA. No momento da indução da ovulação, 6 horas e 24 horas pós IA foram realizadas coletas de material para exame bacteriológico, citológico e para dosagem das citocinas. Neste trabalho foi possível observar que tanto o crescimento de bactérias como a recuperação embrionária obtiveram resultados similares entre os grupos controle e tratado. No entanto, foi observada uma diminuição significativa na contagem de PMNs na citologia uterina 6 e 24 horas pós-cobertura, no grupo tratado com MC (p< 0,05). No que diz respeito às interleucinas, observamos que para a IL-10 não houve diferenças entre os grupos controle e tratado ao longo dos tempos. Porém, o grupo controle apresentou um aumento numérico após inseminação - fato que não ocorreu no grupo tratado. Para o TNF-α também não houve diferenças entre os grupos, mas observou-se um decréscimo numérico nos tempos 6 e 24 horas no grupo tratado. No que se refere a IL-6, foi observada a diferença entre os grupos. No grupo controle houve diminuição nos tempos 6 e 24 horas em comparação ao tempo 0 hora - o que não ocorreu no grupo tratado. Com isso, podemos concluir que o MC foi efetivo na modulação da resposta inflamatória em éguas susceptíveis a EPPC.
Endometritis is the main cause of subfertility in the equine species. This disease is characterized by the accumulation of fluid inside the uterus, influx of polymorphonuclear cells into the uterine lumen, exacerbated expression of pro-inflammatory mediators and, consequently, low fertility rates. In mammals in general, immediately after the semen's contact with the endometrium, an inflammatory response occurs in order to eliminate dead sperm, cellular debris, seminal plasma and microorganisms that are carried at the time of copulation / artificial insemination (AI). Mares can be characterized as resistant or susceptible to endometritis. Resistant mares have the ability to modulate the uterine inflammatory response up to 48 hours after coverage / AI, while susceptible mares do not have this ability. The most commonly used treatments for endometritis are uterine lavages, administration of ecbolic drugs, antibiotics and anti-inflammatory drugs. With the advancement of cell therapy techniques capable of modulating the inflammatory response, different treatments have been studied with promising results in the treatment of endometritis in mares. A modulator of the inflammatory response that has been studied and shows itself as a therapeutic possibility for the treatment of mares with EPPC is the conditioned medium of stem cells (MC). Thus, the present study aimed to review factors related to endometritis in mares and the development of cell therapies, especially stem cells and the conditioned medium (CM) of mesenchymal stem cells. In the present study it was possible to observe that both the growth of bacteria and the embryonic recovery obtained similar results between the control and treated groups. However, a significant decrease in PMN count was observed in uterine cytology 6 and 24 hours post-coverage, in the group treated with MC (p<0.05). Regarding to interleukins, we observed that for IL-10 there were no differences between the control and treated groups over time. However, the control group showed a numerical increase after insemination - a fact that did not occur in the control group. For TNF-α there were also no differences between the groups, but a numerical decrease was observed at times 6 and 24 hours in the treated group. With regard to IL-6, the difference between groups was observed. In the control group, there was a decrease in times 6 and 24 hours compared to time 0 hours, which did not occur in the treated group. With that, we can conclude that the MC was effective in modulating the inflammatory response in mares susceptible to EPPC.
Endometritis is the main cause of subfertility in the equine species. This disease is characterized by the accumulation of fluid inside the uterus, influx of polymorphonuclear cells into the uterine lumen, exacerbated expression of pro-inflammatory mediators and, consequently, low fertility rates. In mammals in general, immediately after the semen's contact with the endometrium, an inflammatory response occurs in order to eliminate dead sperm, cellular debris, seminal plasma and microorganisms that are carried at the time of copulation / artificial insemination (AI). Mares can be characterized as resistant or susceptible to endometritis. Resistant mares have the ability to modulate the uterine inflammatory response up to 48 hours after coverage / AI, while susceptible mares do not have this ability. The most commonly used treatments for endometritis are uterine lavages, administration of ecbolic drugs, antibiotics and anti-inflammatory drugs. With the advancement of cell therapy techniques capable of modulating the inflammatory response, different treatments have been studied with promising results in the treatment of endometritis in mares. A modulator of the inflammatory response that has been studied and shows itself as a therapeutic possibility for the treatment of mares with EPPC is the conditioned medium of stem cells (MC). Thus, the present study aimed to review factors related to endometritis in mares and the development of cell therapies, especially stem cells and the conditioned medium (CM) of mesenchymal stem cells. In the present study it was possible to observe that both the growth of bacteria and the embryonic recovery obtained similar results between the control and treated groups. However, a significant decrease in PMN count was observed in uterine cytology 6 and 24 hours post-coverage, in the group treated with MC (p<0.05). Regarding to interleukins, we observed that for IL-10 there were no differences between the control and treated groups over time. However, the control group showed a numerical increase after insemination - a fact that did not occur in the control group. For TNF-α there were also no differences between the groups, but a numerical decrease was observed at times 6 and 24 hours in the treated group. With regard to IL-6, the difference between groups was observed. In the control group, there was a decrease in times 6 and 24 hours compared to time 0 hours, which did not occur in the treated group. With that, we can conclude that the MC was effective in modulating the inflammatory response in mares susceptible to EPPC.
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Palavras-chave
Endometrite, Equinos, Imunomodulação, Terapia celular, Endometritis, Horses, Immunomodulation, Cell therapy