GLYCOLYTIC AND OXIDATIVE-METABOLISM IN PRIMARY RENAL PROXIMAL TUBULE CULTURES
GLYCOLYTIC AND OXIDATIVE-METABOLISM IN PRIMARY RENAL PROXIMAL TUBULE CULTURES
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DOI：
10.1152/ajpcell.1989.257.2.c333
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发表时间：
1989-08-01
影响因子：
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通讯作者：
MANDEL, LJ
中科院分区：
文献类型：
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作者：
DICKMAN, KG;MANDEL, LJ
培养的细胞在适应培养环境期间经常表现出能量代谢的改变（糖酵解活性增加和氧化代谢减少）。通过比较固定培养皿 (DISH)、摇动锥形瓶 (SHAKE) 和转移回摇动条件 (RESHAKE) 的原代兔肾近曲小管 (RPT) 培养物中的代谢，检查缺氧作为这些效应中介者的作用。 SHAKE 培养物在 24 小时内完全保留了氧化代谢和转运能力。相反，在 6 小时内，DISH 培养物表现出运输依赖性和非依赖性耗氧量、呼吸能力以及 ATP 和 K+ 含量的持续下降。 DISH 培养物中氧化代谢的丧失伴随着乳酸产生的刺激，这在铺板后 1 小时内即可检测到。 DISH 培养物的代谢特性与暴露于分级缺氧水平的 RPT 的代谢特性的比较表明，DISH 培养物中的中等氧张力可能低至 1-3%。 RESHAKE 培养物表现出与 SHAKE 培养物相当的代谢特性，表明 DISH 培养物代谢在复氧时具有可逆性。我们得出的结论是，由于静态培养条件，DISH 培养物会迅速缺氧。振荡悬浮培养物可以为长期体外研究提供代谢更合适的模型。
Cultured cells often exhibit alterations in energy metabolism (increased glycolytic activity and decreased oxidative metabolism) during adaptation to the culture environment. The role of hypoxia as a mediator of these effects was examined by comparison of metabolism in primary rabbit renal proximal tubule (RPT) cultures maintained in stationary culture dishes (DISH), shaking Erlenmeyer flasks (SHAKE), and DISH cultures transferred back to SHAKE conditions (RESHAKE). Both oxidative metabolism and transport capacity were fully preserved in SHAKE cultures over a 24-h period. In contrast, within 6 h, DISH cultures exhibited a continuous decline in transport-dependent and -independent oxygen consumption, respiratory capacity, and ATP and K+ contents. The loss of oxidative metabolism in DISH cultures was accompanied by stimulation of lactate production, detectable within 1 h after plating. Comparison of metabolic properties of DISH cultures to those of RPT exposed to graded levels of hypoxia suggested that medium oxygen tensions may be as low as 1-3% in DISH cultures. RESHAKE cultures exhibited metabolic properties comparable to those of SHAKE cultures, indicating reversibility of DISH culture metabolism on reoxygenation. We concluded that DISH cultures raplidy become hypoxic as a consequence of static culture conditions. Shaking suspension cultures may provide a more metabolically appropriate model for longterm in vitro studies.