Cell Physiology Laboratory, Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, Uttar Pradesh, India.
Cell Physiology Laboratory, Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, Uttar Pradesh, India.
Eur J Pharmacol. 2020 Jul 5;878:173115. doi: 10.1016/j.ejphar.2020.173115. Epub 2020 Apr 14.
The involvement of specific phosphodiesterases (PDEs) in the modulation of cAMP and thereby spontaneous meiotic resumption remains poorly understood. This work aims to evaluate the effects of cilostamide and rolipram (PDE 3A and PDE 4D inhibitors) on spontaneous meiotic resumption from diplotene arrest in rat oocytes cultured in vitro. For this purpose, diplotene-arrested cumulus oocyte complexes (COCs) were collected from rat ovary. The COCs and denuded oocytes were exposed to various concentrations of cilostamide (0.0, 2.5, 5.0 and 10 μM) and rolipram (0, 10, 50 and 100 μM) for various times (0, 3, 5, 7, 14, 16, 18, 20, 22 and 24 h). Cilostamide inhibited spontaneous meiotic resumption in a concentration- and time-dependent manner in COCs and denuded oocytes. Although rolipram showed inhibition of spontaneous meiotic resumption up to some extent, cilostamide was more potent to prevent spontaneous meiotic resumption in both COCs and denuded oocytes. Cilostamide significantly reduced PDE 3A expression, increased cAMP level and prevented spontaneous meiotic resumption in COCs and denuded oocytes. Although rolipram inhibited PDE 4D expression in cumulus cells, increased cAMP level but was not sufficient to prevent spontaneous meiotic resumption. We conclude that both drugs prevent spontaneous resumption from diplotene-arrest through PDE 3A/PDE 4D-cAMP mediated pathway. However, as compare to rolipram, cilostamide was more potent in preventing spontaneous resumption from diplotene-arrest in rat oocytes cultured in vitro. Thus, cilostamide could be used as a potential candidate for the development of female contraceptive drug in future.
特定磷酸二酯酶 (PDEs) 在调节 cAMP 从而自发减数分裂恢复中的作用仍知之甚少。本工作旨在评估西洛司特和罗利普兰（PDE3A 和 PDE4D 抑制剂）对体外培养的大鼠卵母细胞从双线期阻滞自发减数分裂恢复的影响。为此，从大鼠卵巢中收集双线期阻滞的卵丘卵母细胞复合物 (COC)。将 COC 和去卵丘卵母细胞暴露于不同浓度的西洛司特（0.0、2.5、5.0 和 10 μM）和罗利普兰（0、10、50 和 100 μM）不同时间（0、3、5、7、14、16、18、20、22 和 24 h）。西洛司特以浓度和时间依赖的方式抑制 COC 和去卵丘卵母细胞的自发减数分裂恢复。虽然罗利普兰在一定程度上显示出对自发减数分裂恢复的抑制作用，但西洛司特在 COC 和去卵丘卵母细胞中更有效地阻止自发减数分裂恢复。西洛司特显著降低 PDE3A 表达，增加 cAMP 水平，并阻止 COC 和去卵丘卵母细胞的自发减数分裂恢复。虽然罗利普兰抑制卵丘细胞中 PDE4D 的表达，增加 cAMP 水平，但不足以防止自发减数分裂恢复。我们得出结论，这两种药物都通过 PDE3A/PDE4D-cAMP 介导的途径阻止从双线期阻滞自发恢复。然而，与罗利普兰相比，西洛司特在体外培养的大鼠卵母细胞中更有效地阻止从双线期阻滞自发恢复。因此，西洛司特将来可作为开发女性避孕药的潜在候选药物。
