Changes in cell culture temperature alter release of inflammatory mediators in murine macrophagic RAW264.7 cells
Changes in cell culture temperature alter release of inflammatory mediators in murine macrophagic RAW264.7 cells
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DOI：
10.1007/s00011-007-6161-z
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发表时间：
2007-07-01
影响因子：
6.7
通讯作者：
Noguchi, T.
中科院分区：
文献类型：
--
作者：
Hagiwara, S.;Iwasaka, H.;Noguchi, T.
目的：检查细胞培养温度的适度变化是否会影响各种细胞因子和相关炎症介质的产生。方法：我们在高温（40°C）、常温（37°C）和低温（34°C）条件下对小鼠巨噬细胞RAW264.7细胞系进行脂多糖（LPS）刺激。然后，我们测量了细胞中热休克蛋白 70 (HSP70)、热休克因子蛋白 (HSF) 和核因子-kB (NF-kB) 二聚体（p50 和 p65）的水平，以及培养物中高迁移率族蛋白 1 (HMGB1) 和细胞因子肿瘤坏死因子-α (TNF-α)、白细胞介素 1 β (IL-1 β) 和白细胞介素 6 (IL-6) 的水平。结果：与37℃相比，40℃和34℃LPS刺激后，HMGB1、IL-1β、IL-6和TNF-α以及NF-kB二聚体（p50和p65）水平均降低。HSP70和HSF水平在40℃和34℃时升高。 LPS 诱导细胞激活后的低温可减弱炎症反应并降低细胞损伤的可能性。这些发现表明，适度的温度变化可以调节炎症反应，可能是对抗脓毒症的有效疗法。
Objectives: To examine whether moderate changes in cell culture temperature influence the production of various cytokines and associated mediators of inflammation.Methods: We performed lipopolysaccharide (LPS) stimulation of the murine macrophagic RAW264.7 cell line under hyperthermic (40 degrees C), normothermic (37 degrees C) and hypothermic (34 degrees C) conditions. We then measured the levels of heat shock protein 70 (HSP70), heat shock factor protein (HSF) and nuclear factor-kB (NF-kB) dimers (p50 and p65) in the cells, and the levels of high mobility group box 1 (HMGB1) and the cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin 1 beta (IL-1 beta) and interleukin 6 (IL-6) in the culture supernatants.Results: Levels of HMGB1, IL-1 beta, IL-6, and TNF-alpha, as well as NF-kB dimers (p50 and p65), were all reduced following LPS stimulation at 40 degrees C and 34 degrees C compared with those at 37 degrees C. Levels of HSP70 and HSF increased at 40 degrees C and 34 degrees C.Conclusions: The application of moderate hyperthermia and hypothermia after LPS-induced cell activation attenuated the inflammatory response and reduced the likelihood of cell damage. These findings suggest that moderate temperature changes modulate the inflammatory response and could be a useful therapy against sepsis.