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目的:研究牛心内膜内皮细胞(BEEC)腺苷三磷酸双磷酸酶(apyrase)的特性并比较牛心内膜及血管内皮细胞的胞外腺苷酸酶活性。方法:以反相HPLC测定核苷酸含量,以磷离子释放法检测apyrase活性。结果:叠氮钠10mmol·L~(-1)和氟化钠20mmol·L~(-1)分别抑制BEEC apyrase 51％及38％活性,而Na~+/K~+ -ATP酶抑制剂哇巴因则对BEEC apyrase活性无影响。过氧化氢0.5mmol·L~(-1)呈时间依赖性地抑制心内膜内皮细胞apyrase活性。BEEC的apyrase活性低于主动脉内皮细胞的apyrase活性,而BEEC的胞外AMP酶则远较血管内皮细胞的活性高。结论:BEEC apyrase具有叠氮钠和氟化钠敏感的、哇巴因不敏感的特性;与血管内皮细胞相比,BEEC apyrase活性较高而胞外AMP酶活性较低。
Objective: To study the characteristics of apyrase in bovine endocardial endothelial cells (BEECs) and to compare the extracellular adenylase activity of bovine endocardial and vascular endothelial cells. Methods: The content of nucleotides was determined by reverse phase HPLC, and the activity of apyrase was detected by phosphorus ion release method. Results: Sodium azide 10 mmol·L -1 and sodium fluoride 20 mmol·L -1 inhibited BEEC apyrase activity by 51% and 38% respectively, while Na ~ + / K ~ + -ATPase inhibitors Pakistan has no effect on BEEC apyrase activity. Hydrogen peroxide 0.5 mmol·L -1 inhibited the apyrase activity of endocardial endothelial cells in a time-dependent manner. The apyrase activity of BEEC is lower than that of aortic endothelial cells, whereas the extracellular AMP enzyme of BEEC is far more active than that of vascular endothelial cells. CONCLUSION: BEEC apyrase possesses the properties of sodium azide and sodium fluoride sensitive, ouabain-insensitive. Compared with vascular endothelial cells, BEEC apyrase activity is higher and extracellular AMP enzyme activity is lower.