Avaliação da espermatogênese e da imunoexpressão de proteínas do epitélio seminífero e de citocinas em testículos de ratos tratados com anti-inflamatório
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2023-12-15
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Universidade Estadual Paulista (Unesp)
Resumo
A diacereína, um fármaco anti-inflamatório utilizado no tratamento da osteoartrite, inibe a ação de citocinas, como o fator de necrose tumoral-alfa (TNF-α) e interleucina-1β (IL-1), as quais atuam em condições fisiológicas em vários órgãos, inclusive nos testículos. Considerando a escassa literatura sobre a participação de citocinas em condições fisiológicas no epitélio seminífero, foi proposto avaliar se a deficiência de citocinas, induzida pela diacereína, afeta a espermatogênese e as células de Leydig (CL). Foram utilizados 16 ratos machos adultos, distribuídos em 2 grupos: grupo controle (GC; n=8) e grupo diacereína (GD; n=8). Os animais do GC receberam solução salina e os animais do GD receberam 100 mg/kg p.c. de diacereína por gavagem, por 30 dias. Os testículos foram coletados, fixados e processados para inclusão em historesina e parafina para as análises morfológicas, morfométricas e imuno-histoquímicas. Nos cortes corados pela H.E., o número de espermatócitos (SP) foi quantificado, e imunorreações foram realizadas para avaliar o número de células germinativas Ki-67-positivas (marcador de divisão celular), macrófagos recém-chegados (CD68/ED1) e a imunoexpressão de citocinas pró-inflamatórias (IL-1β e TNF-α), conexina 43 (junção entre células do epitélio), vimentina (marcador de célula de Sertoli), bem como a dupla marcação de 17β-HSD7 (marcador de CL) + IL-1β nas CL. Os dados foram submetidos ao test t de Student (p≤0.05). No GD, a imunoexpressão de IL-1β e TNF-α reduziram, confirmando a deficiência destas citocinas no testículo. Os níveis séricos de testosterona reduziram significantemente em associação à significante redução de CL positivas à IL-1β, confirmando que essas CL são vulneráveis ao tratamento com diacereína. Os túbulos seminíferos (TS) apresentaram células descamadas na luz tubular, redução no número de SP, de células Ki-67-positivas, bem como redução na imunoexpressão de proteínas estruturais (conexina 43 e vimentina) e de TNF-α. Ademais, houve aumento no número de macrófagos recém-chegados (CD68/ED1) no tecido intersticial. Considerando que o epitélio seminífero é andrógeno-dependente e responsivo aos níveis de TNF-α, a redução dos níveis de testosterona e de TNF-α poderiam explicar as alterações verificadas nos TS, inclusive a redução de SP, de conexina 43 e a baixa atividade mitótica/meiótica. Portanto, a deficiência de citocinas, induzida pelo tratamento com diacereína, prejudica tanto a esteroidogênese quanto a espermatogênese em condições normais. Uma atenção especial deve ser dada ao perfil hormonal reprodutivo de pacientes tratados com anti-inflamatórios por longos períodos.
Diacerein, an anti-inflammatory drug used in the treatment of osteoarthritis, inhibits the action of cytokines, such as tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1), which act in physiological conditions in various organs, including the testicles. Considering the scarce literature on the participation of cytokines in physiological conditions in the seminiferous epithelium, it was proposed to evaluate whether cytokine deficiency, induced by diacerein, affects spermatogenesis and Leydig cells (LC). Sixteen adult male rats were used, divided into 2 groups: control group (GC; n=8) and diacerein group (GD; n=8). The CG animals received saline solution and the DG animals received 100 mg/kg p.c. of diacerein by gavage, for 30 days. Testes were collected, fixed and processed for inclusion in historesin and paraffin for morphological, morphometric and immunohistochemical analyses. In H.E. stained sections, the number of spermatocytes (SP) was quantified, and immunoreactions were performed to assess the number of Ki-67-positive germ cells (cell division marker), newly arrived macrophages (CD68/ED1) and the immunoexpression of pro-inflammatory cytokines (IL-1β and TNF-α), connexin-43 (junction between epithelial cells), vimentin (Sertoli cell marker), as well as the double labeling of 17β-HSD7 (CL marker) + IL-1β in LC. Data were submitted to Student's t test (p≤0.05). In GD, the immunoexpression of IL-1β and TNF-α decreased, confirming the deficiency of these cytokines in the testis. Serum testosterone levels significantly reduced in association with the significant reduction of IL-1β positive CL, confirming that these CL are vulnerable to treatment with diacerein. The seminiferous tubules (TS) showed desquamated cells in the tubular lumen, reduced number of SP and Ki-67-positive cells, as well as reduced immunoexpression of structural proteins (connexin 43 and vimentin) and TNF-α. Furthermore, there was an increase in the number of newly arrived macrophages (CD68/ED1) in the interstitial tissue. Considering that the seminiferous epithelium is androgen-dependent and responsive to TNF-α levels, the reduction in testosterone and TNF-α levels could explain the changes observed in TS, including the reduction in SP, connexin 43 and the low mitotic/meiotic activity. Therefore, cytokine deficiency induced by diacerein treatment impairs both steroidogenesis and spermatogenesis under normal conditions. Special attention should be given to the reproductive hormone profile of patients treated with anti-inflammatory drugs for long periods.
Diacerein, an anti-inflammatory drug used in the treatment of osteoarthritis, inhibits the action of cytokines, such as tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1), which act in physiological conditions in various organs, including the testicles. Considering the scarce literature on the participation of cytokines in physiological conditions in the seminiferous epithelium, it was proposed to evaluate whether cytokine deficiency, induced by diacerein, affects spermatogenesis and Leydig cells (LC). Sixteen adult male rats were used, divided into 2 groups: control group (GC; n=8) and diacerein group (GD; n=8). The CG animals received saline solution and the DG animals received 100 mg/kg p.c. of diacerein by gavage, for 30 days. Testes were collected, fixed and processed for inclusion in historesin and paraffin for morphological, morphometric and immunohistochemical analyses. In H.E. stained sections, the number of spermatocytes (SP) was quantified, and immunoreactions were performed to assess the number of Ki-67-positive germ cells (cell division marker), newly arrived macrophages (CD68/ED1) and the immunoexpression of pro-inflammatory cytokines (IL-1β and TNF-α), connexin-43 (junction between epithelial cells), vimentin (Sertoli cell marker), as well as the double labeling of 17β-HSD7 (CL marker) + IL-1β in LC. Data were submitted to Student's t test (p≤0.05). In GD, the immunoexpression of IL-1β and TNF-α decreased, confirming the deficiency of these cytokines in the testis. Serum testosterone levels significantly reduced in association with the significant reduction of IL-1β positive CL, confirming that these CL are vulnerable to treatment with diacerein. The seminiferous tubules (TS) showed desquamated cells in the tubular lumen, reduced number of SP and Ki-67-positive cells, as well as reduced immunoexpression of structural proteins (connexin 43 and vimentin) and TNF-α. Furthermore, there was an increase in the number of newly arrived macrophages (CD68/ED1) in the interstitial tissue. Considering that the seminiferous epithelium is androgen-dependent and responsive to TNF-α levels, the reduction in testosterone and TNF-α levels could explain the changes observed in TS, including the reduction in SP, connexin 43 and the low mitotic/meiotic activity. Therefore, cytokine deficiency induced by diacerein treatment impairs both steroidogenesis and spermatogenesis under normal conditions. Special attention should be given to the reproductive hormone profile of patients treated with anti-inflammatory drugs for long periods.
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anti-inflamatório, células germinativas, conexina 43, germ cells, anti-inflammatory, connexin 43
Como citar
JESUS, Elide Loise Freitas de. Avaliação da Espermatogênese e da Imunoexpressão de Proteínas do Epitélio Seminífero e de Citocinas em Testículos de Ratos Tratados com Anti-inflamatório. Orientador: Estela Sasso Cerri. 2024. Trabalho de Conclusão de Curso (Bacharelado em Farmácia-Bioquímica) - Faculdade de Ciências Farmacêuticas - UNESP Araraquara, UNESP, 2023. p. 61.