Cazin M, Paluszezak D, Poulain V, Dine T, Bianchi A, Cazin J C, Aerts C
Laboratoire de Pharmacologie, Faculté de Pharmacie, Lille, France.
Methods Find Exp Clin Pharmacol. 1988 Apr;10(4):231-7.
Alveolar macrophages are able to adapt their energy metabolism to very difficult survival conditions. Gaseous phase culture is adaptable to alveolar macrophages because it reproduces in vitro conditions very similar to in vivo conditions. It is easy to modify the incubation gas composition for hypoxia and anaerobiosis. Metabolic changes and cell injury were evaluated in three studies carried out after 24 hr of gaseous phase culture in normoxia and in anaerobiosis with a possible treatment with 0.01 microgram/ml vincamine: 1) ATP content assay by bioluminescence, the witness of cell vitality which decreases significantly in anaerobiosis; 2) Lactate assay which shows the metabolism derivation towards the anaerobic pathways; and 3) Tritiated deoxyglucose (DOG) incorporation, which shows glucose requirements after hypoxic incubation, maintaining or recovering a certain level of energetic activity. This incorporation greatly increases after anaerobic culture. Vincamine has no activity in normoxia. The three parameters are not significantly different from control, but in anaerobiosis, vincamine reveals an interesting protective effect. ATP content decreases under treatment and DOG incorporation increases. This demonstrates that vincamine is able to maintain cell metabolic activity for a longer period of time after the beginning of hypoxic trial. Cells can better use their energy storage and the metabolic pathways which enable them to restore themselves, thanks to vincamine treatment. It has been shown that cell membrane integrity was preserved by tests using cytochalasin B. DOG was not incorporated by cells treated with cytochalasin B after 24 hr of anaerobic culture and normally incorporated by control cells.(ABSTRACT TRUNCATED AT 250 WORDS)
肺泡巨噬细胞能够使其能量代谢适应非常恶劣的生存条件。气相培养适用于肺泡巨噬细胞,因为它能在体外再现与体内条件非常相似的环境。改变培养气体成分以模拟缺氧和无氧环境很容易。在常氧和无氧条件下进行24小时气相培养后,开展了三项研究,评估代谢变化和细胞损伤情况,研究中可能用0.01微克/毫升长春胺进行处理:1)通过生物发光法检测ATP含量,这是细胞活力的指标,在无氧环境中会显著下降;2)检测乳酸含量,以显示代谢向无氧途径的转变;3)检测氚标记脱氧葡萄糖(DOG)的掺入情况,以显示缺氧培养后的葡萄糖需求,维持或恢复一定水平的能量活性。在无氧培养后,这种掺入量会大幅增加。长春胺在常氧条件下无活性。这三个参数与对照组无显著差异,但在无氧环境中,长春胺显示出有趣的保护作用。处理后ATP含量下降,DOG掺入量增加。这表明长春胺能够在缺氧试验开始后更长时间维持细胞代谢活性。由于长春胺的处理,细胞能够更好地利用其能量储备以及使其自我恢复的代谢途径。通过使用细胞松弛素B的试验表明,细胞膜完整性得以保留。在无氧培养24小时后,用细胞松弛素B处理的细胞不掺入DOG,而对照细胞正常掺入DOG。(摘要截取自250词)