Effect of culture PO2 on macrophage (RAW 264.7) nitric oxide production
Effect of culture PO2 on macrophage (RAW 264.7) nitric oxide production
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DOI：
10.1152/ajpcell.2001.280.2.c280
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发表时间：
2001-02-01
影响因子：
5.5
通讯作者：
Baumgardner, JE
中科院分区：
文献类型：
--
作者：
Otto, CM;Baumgardner, JE
巨噬细胞通常在 149 Torr 的 Po-2 下培养，但体内组织巨噬细胞生活在氧张力低得多的环境中。尽管氧张力变化影响一氧化氮 (NO) 合成的潜在机制有很多，但很少有报道研究 Po-2 对巨噬细胞 NO 产生的影响。通过使用严格控制氧张力的培养室，我们研究了培养物 Po-2 对巨噬细胞 NO 产生、诱导型一氧化氮合酶 (iNOS) 活性、iNOS 蛋白和肿瘤坏死因子产生的影响。 NO 产生和 iNOS 活性在 39.4 至 677 Torr 范围内呈线性相关，但在 1.03 至 39.4 Torr 范围内不呈线性相关。因此，细胞培养中常用的高氧张力体外获得的结果与在较低氧张力下培养的细胞获得的结果在数量和质量上有所不同，后者更准确地反映了体内环境。氧张力对 NO 产生的影响对细胞培养方法以及脓毒症啮齿动物模型中微循环功能障碍与炎症反应之间的关系具有影响。
Macrophages are commonly cultured at a Po-2 of 149 Torr, but tissue macrophages in vivo live in an environment of much lower oxygen tension. Despite the many potential mechanisms for changes in oxygen tension to influence nitric oxide (NO) synthesis, there have been few reports investigating the effect of Po-2 on macrophage NO production. With the use of a culture chamber designed to rigorously control oxygen tension, we investigated the effects of culture Po-2 on macrophage NO production, inducible nitric oxide synthase (iNOS) activity, iNOS protein, and tumor necrosis factor production. NO production and iNOS activity were linearly related in the range of 39.4 to 677 Torr, but not in the range of 1.03 to 39.4 Torr. Therefore, results obtained in vitro for the high oxygen tensions commonly used in cell culture were quantitatively and qualitatively different from results obtained in cells cultured at the lower oxygen tensions that more accurately reflect the in vivo environment. The influence of oxygen tension on NO production has implications for cell culture methodology and for the relationship between microcirculatory dysfunction and inflammatory responses in rodent models of sepsis.