Schini V, Grant N J, Miller R C, Takeda K
Université Louis Pasteur, Laboratoire de Pharmacologie Cellulaire et Moléculaire, CNRS UA 600, Strasbourg/France.
Eur J Cell Biol. 1988 Oct;47(1):53-61.
Primary, first and second passaged endothelial cells from bovine aorta were grown in plastic culture dishes or on glass coverslips. The cells were characterized by their monolayer cobblestone appearance at confluence, their immunofluorescent staining for factor VIII-related antigen, their specific uptake of low density lipoprotein and by their ultrastructure. Following stimulation of the cells by atriopeptin II or sodium nitroprusside, both cellular and extracellular cyclic GMP levels were measured. Cellular cyclic GMP content was increased greatly by atriopeptin II in a time-dependent manner while sodium nitroprusside was essentially without effect. Increases in tissue cyclic GMP levels were associated with a time-dependent accumulation of the nucleotide in the extracellular compartment. Zaprinast, a specific inhibitor of cyclic GMP phosphodiesterases, did not significantly affect either basal or atriopeptin II-stimulated increases in cyclic GMP content, nor extracellular accumulation of the nucleotide. It is concluded that the cyclic GMP content of endothelial cells is not solely dependent on degradation by phosphodiesterases but also involves release of cyclic GMP into the extracellular compartment.
取自牛主动脉的原代、传代1次和传代2次的内皮细胞，分别培养于塑料培养皿或玻璃盖玻片上。这些细胞的特征包括：汇合时呈单层鹅卵石样外观、对VIII因子相关抗原进行免疫荧光染色、特异性摄取低密度脂蛋白以及具有特定的超微结构。在用心房肽II或硝普钠刺激细胞后，对细胞内和细胞外的环磷酸鸟苷（cGMP）水平进行了测定。心房肽II能使细胞内cGMP含量随时间依赖性大幅增加，而硝普钠基本无此作用。组织cGMP水平的升高与细胞外区室中该核苷酸的时间依赖性积累有关。环磷酸鸟苷磷酸二酯酶的特异性抑制剂扎普司特，对基础cGMP含量或心房肽II刺激引起的cGMP含量增加均无显著影响，对细胞外核苷酸的积累也无影响。由此得出结论，内皮细胞的环磷酸鸟苷含量不仅取决于磷酸二酯酶的降解，还涉及环磷酸鸟苷释放到细胞外区室。
