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新生大鼠培养心肌细胞中腺苷的形成与释放

Adenosine formation and release from neonatal-rat heart cells in culture.

作者信息

Meghji P, Holmquist C A, Newby A C

出版信息

Biochem J. 1985 Aug 1;229(3):799-805. doi: 10.1042/bj2290799.

Abstract

The incorporation of [3H]adenosine (10 microM) into neonatal-rat heart cell nucleotides was inhibited in a concentration-dependent manner, such that 50% inhibition was obtained with 0.75 microM-dipyridamole, 0.26 microM-hexobendine or 0.22 microM-dilazep. Adenosine formation was accelerated 2.5-fold to 2.1 +/- 0.3 nmol/10(7) cells in 10 min when cells were incubated with a combination of 30 mM-2-deoxyglucose and 2 micrograms of oligomycin/ml. Of the newly formed adenosine, 6 +/- 2% was in the cells. Dipyridamole, hexobendine or dilazep (10 microM) increased the amount of adenosine in the cells and decreased that in the medium such that 45-50% of the newly formed adenosine was in the cells. Antibodies which inhibited ecto-5'-nucleotidase by 98.7 +/- 0.3% did not alter the rate of adenosine formation or its distribution between cells and medium. We conclude that adenosine was formed in the cytoplasm during catabolism of cellular ATP and was released via the dipyridamole-sensitive symmetric nucleoside transporter.

摘要

[3H]腺苷(10微摩尔)掺入新生大鼠心脏细胞核苷酸的过程受到浓度依赖性抑制,即0.75微摩尔双嘧达莫、0.26微摩尔己酮可可碱或0.22微摩尔地拉卓可产生50%的抑制作用。当细胞与30毫摩尔2-脱氧葡萄糖和每毫升2微克寡霉素共同孵育时,10分钟内腺苷生成加速2.5倍,达到2.1±0.3纳摩尔/10⁷个细胞。新生成的腺苷中,6±2%存在于细胞内。双嘧达莫、己酮可可碱或地拉卓(10微摩尔)可增加细胞内腺苷量,减少培养基中的腺苷量,使得新生成的腺苷有45 - 50%存在于细胞内。抑制胞外5'-核苷酸酶达98.7±0.3%的抗体并未改变腺苷生成速率及其在细胞与培养基之间的分布。我们得出结论,腺苷在细胞ATP分解代谢过程中于细胞质内生成,并通过对双嘧达莫敏感的对称核苷转运体释放。

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