三氟啦嗪对视网膜色素上皮细胞cAMP-PKA信号系统的影响

目的 :检测TFP对培养的豚鼠RPE细胞cAMP浓度和PKA活性的影响。探讨CaM被抑制时 ,与cAMP -PKA信号系统相关性及在RPE细胞增殖调控中的作用。从信号传导角度探讨TFP抑制RPE细胞迁移和增殖的机制。方法 :1 应用12 5I -cAMP试剂盒 ,采用放免法测定 10umol/LTFP作用不同时间RPE细胞内cAMP浓度的变化。 2 应用液闪法测定 10umol/LTFP作用不同时间后 ,RPE细胞PKA活性的变化。结果 :1 TFP对豚鼠RPE细胞cAMP浓度的影响 :统计结果表明 ,与对照组cAMP值 ( 0 410 9± 0 0 931pmol/5 0ul)相比 ,TFP作用 5分钟内即引起cAMP快速的升高达 ( 0 80 5 1± 0 5 5 3pmol/5 0ul) (P <0 0 1) ,直到 3小时仍维持较高的水平 ( 0 72 85±0 0 2 94pmol/5 0ul) (P <0 0 5 )。 2 TFP对豚鼠RPE细胞PKA活性的影响 :统计结果表明 ,与对照组PKA活性值 ( 4 5 5 73±2 41cpm/mgprotiein)相比 ,TFP作用 5min内即引起PKA活性快速上升达 ( 135 6 5± 1 81cpm /mg protein) (P <0 0 1) ,直至3h仍维持较高水平 ( 6 5 95± 5 0 1cpm/mg protein) (P <0 0 5 )。结论 :1 在TFP抑制RPE细胞增殖过程中 ,cAMP -PKA信号系统呈上调趋势。 2 在RPE细胞增殖抑制过程中 ,Ca2 + -CaM系统与cAMP -PKA信号系统存在着拮抗关系。 3 在TFP引起cA

Trifluoperazine Effect On cAMP- PKA Message Systems In Retinal Pigment Epithelial Cells

Objective:To study the proliferative inhibition of Trifluoperazine(TFP)and its effect on the cAMP- protein kinase A(cAMP-PKA)signal system in cultured guinea pig RPE cell. Methods:10umol/L TFP were added to cultured RPE cells.The intracellular cAMP concentration and PKA activity were measured by Radioimmunoassay. Results:The intracellular cAMP level was higher than that of the controls after treated with 10umol/L TFP for 5 minutes and kept at high level till 3 hours.Meanwhile the activity of PKA(cAMP dependent protein kinase A)increased within 5 minutes and kept high till 3 hours. Conclusions:The results indicated that the TFP could increase the level and activity of the cAMP-PKA signal system in cultured guinea pig RPE cell.