Assessing physiological concentrations of endogenous substances in situ by inducing calcium oscillations in vitro. Case of liver^1  

利用体外诱发的钙振荡估测在体内源性物质的生理浓度.肝脏中的情况(英文)

作　　者：崔宗杰[1] 郭丽丽[1] 

机构地区：[1]北京师范大学细胞生物学研究所,北京中国100875

出　　处：《Acta Pharmacologica Sinica》2002年第1期29-34,共6页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China,No 39825112,39870367,30070286

摘　　要：AIM:To identify the Physiological concentration ranges of norepinephrine（NE），vasopressin（VP），and ATP in the rat liver.METHODS:Rat hepatocytes were isolated by collagenase pefusion .lsolated cells were loaded with the fluorescent calcium indicator Fum-2 acetoxymethyl ester （Fura-2 AM）.The effects of different concentrations of norepinephrine，vasopressin，and ATP on intracellular calcium concentration ([Ca²⁺])increases in the freshly isolated rat hepatocytes were investigated.[Ca²⁺];was measured by micro-fluorometry and recorded as fluorescence ration(F₃₄₀/F₃₈₀).RESULTS:NE,VP,and ATP induced increases in [Ca²⁺];in a concentration-dependent manner.At lower concentrations,[Ca²⁺];tended to show an oscillatory increase with increasing concentra-tions,[Ca²⁺];in more cells tended to show phasic or plateau increases.NE,VP,and ATP concentrations likely to induce an oscillatory [Ca²⁺];response were 100-500 nmol/L.50-100 pmol/L,and【1чmol/L respectively.CONCLUSION:Physiological concentra-tions of NE,VP,and ATP are 100-500 nmol/L,50-tions，[Ca²⁺];in more cells tended to show phasic or plateau increases.NE,VP,and ATP concentrations likely to induce an oscillatory [Ca²⁺];response were 100-500 nmol/L,50-100 pmol/L,and【1 чmol/L respectively.CONCLUSION:Physiological concentra-tions of NE,VP,and ATP are 100-500 nmol/L,50-100 pmol/L,and【1чmol/L respectively in the rat liver.目的:研究大鼠肝中去甲肾上腺素、加压素、ATP的生理浓度.方法:肝细胞通过胶原水解酶灌流肝脏得到.分离的肝细胞加载钙离子荧光指示探针Fura-2,通过显微荧光术测量胞内钙浓度的变化,以荧光比例(F_(340)/F_(280))表示,分别检测不同浓度范围的去甲肾上腺素、加压素、ATP对单个肝细胞胞浆钙离子浓度升高的作用.结果:去甲肾上腺素、加压素、ATP以剂量依赖的方式诱导胞内钙离子浓度的增加.较低浓度刺激,胞内钙浓度呈振荡性增加;随着刺激物浓度的增加,更多的细胞胞内钙离子浓度呈现缓慢或高台样增加 去甲肾上腺素、加压素、ATP诱发钙振荡的最佳浓度分别是100—500nmol/L、50-100pmol/L和<1μmol/L.结论:在大鼠肝脏中,去甲肾上腺素、加压素、ATP的生理性浓度分别在100-500nmol/L、50—100pmol/L和<1μmol/L范围.

关 键 词：calcium HEPATOCYTES NOREPINEPHRINE VASOPRESSINS adenosine triphosphate rats 

分 类 号：R33[医药卫生—人体生理学]