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目的:研究蛋白激酶C(PKC)在孕酮激发豚鼠精子顶体反应中的作用.方法:豚鼠精子在Ca~(2+)缺乏MCM培养液中,38.5℃5％CO_2/空气获能培养(6-6.5)h然后经Percoll密度梯度离心,再混悬于MCM中,调数至5×10~9个/升.加入PKC激活剂或抑制剂以及相关试剂15分钟后,用相差显微镜评价AR(以AR％表示).结果:在钙离子存在下,PDB或OAG可激发豚鼠精子AR,并可显著提高孕酮激发豚鼠精子AR的作用.然而,STA不仅可抑制AR,也可阻止孕酮激发豚鼠精子AR.同时,依他酸和硝苯地平均可阻断P_4以及P_4和PDB激发豚鼠精子AR.此外,新霉素可抑制P_4激发豚鼠精子AR,而且PDB可部分逆转这种作用.结论:激活或抑制PKC可明显增加或降低P_4激发的豚鼠精子AR,表明PKC在生理性AR的信号转导通路中起重要作用.
Objective: To study the role of protein kinase C (PKC) in progestin-induced guinea pig sperm acrosome reaction.Methods: Guinea pig sperm were cultured in Ca 2+ -free MCM medium at 38.5 ℃ with 5% CO 2 / 6-6.5) h followed by Percoll density gradient centrifugation, resuspended in MCM, adjusted to a number of 5 × 10 ~ 9 / L. After adding PKC activator or inhibitor and related reagents for 15 minutes, phase contrast microscopy AR (AR%) .Results: In the presence of calcium ions, PDB or OAG can stimulate guinea pig sperm AR, and can significantly improve the progesterone-induced guinea pig sperm AR role.However, STA can not only inhibit AR, but also progesterone Eosin and nifedipine blocked both P_4 and P_4 and PDB to stimulate sperm AR in guinea pigs.In addition, neomycin inhibited P_4-induced guinea pig sperm AR, and PDB partially reversed this effect. CONCLUSION: Activation or inhibition of PKC can significantly increase or decrease the P_4-stimulated guinea pig sperm AR, indicating that PKC plays an important role in physiological AR signal transduction pathway.