Effect of protein kinase C on guinea pig sperm acrosome reaction induced by progesterone  

蛋白激酶C在孕酮激发的豚鼠精子顶体反应中的作用(英文)

作　　者：陈文颖[1] 袁玉英[1] 石其贤[1] 张信岳[1] 

机构地区：[1]浙江省医学科学院

出　　处：《Acta Pharmacologica Sinica》2000年第9期21-25,共5页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China (No 39870364);Natural Science Foundation of Zhejiang Province (No 398028);National "9.5" Pan Deng Project.

摘　　要：AIM; To investigate the effect of protein kinase C (PKC) on the guinea pig sperm acrosome reaction (AR) induced by progesterone (P4). METHODS; Guinea pig spermatozoa were preincubated in MCM-Ca2+ DF medium at 38.5℃ under 5 % CO2/air for 6-6.5 h for capacitation, and were then washed through a three-step gradient Percoll, resuspended in MCM-Ca2+ DF and ad-justed to 5×109 cells/L. The AR was assessed by means of phase-contrast microscopy (expressed as percent of AR) 15 min after different reagents were added. RE-SULTS: In the presence of Ca2+ 2 mmol/L, phorbol 12,13-dibutyrate (PDB) or the permanent synthetic dia-cylglycerol, l-oleoy1-2-acetyl-sn-glycerol (OAG) initiat-ed the AR of spermatozoa in a concentration-related man-ner, whereas staurosporine (STA) exhibited an inhibition of the AR. PDB and OAG significantly enhanced P4-in-duced AR that was greater than that treated with either ag-onist separately. STA markedly inhibited the P4-induced AR. The AR induced by PDB was completely antago-nized by inclusion of egtazic acid. Also, both egtazic acid and nifedipine antagonized the AR induced by P4 and PDB + P4 In addition, neomycin also prevented the P4-induced AR. This effect could be partly reversed by PDB. CONCLUSION: Activation or inhibition of PKC significantly plays an important role in the signal trans-duction pathway for the physiological AR.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.

关 键 词：protein kinase C acrosome reaction PROGESTERONE phorbol 12 13-dibutyrate DIACYLGLYCEROL calcium SPERMATOZOA 

分 类 号：R339.2[医药卫生—人体生理学]