摘要
利用培养新生大鼠心肌细胞 ,检测NO前体L -精氨酸 (L -arginine ,L -Arg)和NO供体硝普钠 (sodiumnitroprus side ,SNP)对PKC活性的影响 ,并探讨内、外源性NO在PKC激动剂佛波指 (phorbol 12 -myristate 13-acetate ,PMA)激活PKC中的作用 .实验结果表明 :培养基中加入L -Arg ,PKC活性呈剂量依赖性降低 ;用L -Arg进行预处理 ,30min后加入PMA ,PKC活性明显降低 ,与单纯PMA组相比有显著差异 ;NOS抑制剂L -NAME本身对基础状态PKC活性无明显影响 ,但可阻断L -Arg对上述 2个效应的影响 ;培养液中加入NO供体SNP ,PKC活性呈剂量依赖性降低 ;用SNP预处理心肌细胞 ,5min后加入PMA ,PKC活性与单纯PMA组相比有显著性差异 .以上结果表明 ,内、外源性NO均具有剂量依赖性抑制PKC活性的作用 ,PKC可能是NO对心肌细胞作用的胞内信号传导通路的关键部位或重要信号分子之一 ;L -Arg通过NOS先生成NO ,NO再对PKC起抑制作用 .
The effect of endogenous and exogenous nitric oxide(NO) on protein kinase(PKC) activity in cultured neonatal rat cardiomyocytes was examined.The results were as following:Activity of PKC was decreased by NO precursor L-arginine(L-Arg) (10 -5~10 -2 mol/L,30 min) and NO donor sodium nitroprusside (SNP)(10 -6~10 -3 mol/L,5 min)in a dose-dependent manner in cultured neonatal rat cardiomyocytes.Activity of PKC increased by a PKC activator phorbol 12-myristate 13-acetate(PMA)(10 -5mol/L,15 min) was impaired by a NO precursor L-Arg and a exogenous NO donor SNP.NG-nitro-L-argingie methyl ester(L-NAME),a nitric oxide synthase (NOS) blocker,itself had no effect on the PKC activity of cultured neonatal rat cardiomyocytes,but may inhibite the role of PKC activity induced PMA.These results indicate that PKC was decreased by endogenous and exogenous NO in a dose-dependent manner.The role of NO on rat cardiomyocytes intracellular signaling transduction pathway may be controlled through PKC.PKC may turn into key site or important signaling element of this effect.
出处
《吉首大学学报(自然科学版)》
CAS
2002年第3期32-35,46,共5页
Journal of Jishou University(Natural Sciences Edition)
基金
海南省自然科学基金资助项目 (30 0 18)
海南省卫生厅科研基金资助课题 (琼卫 2 0 0 0 - 78)
关键词
心肌细胞培养
一氧化氮
蛋白激酶C
佛波酯
neonatal rat myocardial cell culture
nitric oxide
protein kinase C
phorbol 12-myristate 13-acetate