Hagiwara S, Iwasaka H, Matsumoto S, Noguchi T
Department of Brain and Nerve Science, Anesthesiology, Oita University Faculty of Medicine, Oita Prefecture, 1-1 Idaigaoka-Hasamamachi-Yufu City, Oita 879-5593, Japan.
Inflamm Res. 2007 Jul;56(7):297-303. doi: 10.1007/s00011-007-6161-z.
To examine whether moderate changes in cell culture temperature influence the production of various cytokines and associated mediators of inflammation.
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 1beta (IL-1beta) and interleukin 6 (IL-6) in the culture supernatants.
Levels of HMGB1, IL-1beta, 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.
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.
研究细胞培养温度的适度变化是否会影响多种细胞因子及相关炎症介质的产生。
我们在高温（40摄氏度）、正常体温（37摄氏度）和低温（34摄氏度）条件下，对小鼠巨噬细胞RAW264.7细胞系进行脂多糖（LPS）刺激。然后我们测量细胞中热休克蛋白70（HSP70）、热休克因子蛋白（HSF）和核因子-κB（NF-κB）二聚体（p50和p65）的水平，以及培养上清液中高迁移率族蛋白B1（HMGB1）和细胞因子肿瘤坏死因子-α（TNF-α）、白细胞介素1β（IL-1β）和白细胞介素6（IL-6）的水平。
与37摄氏度时相比，在40摄氏度和34摄氏度下进行LPS刺激后，HMGB1、IL-1β、IL-6和TNF-α的水平以及NF-κB二聚体（p50和p65）均降低。HSP70和HSF的水平在40摄氏度和34摄氏度时升高。
LPS诱导细胞活化后应用适度的高温和低温可减轻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调节炎症反应，可能是治疗脓毒症的一种有效疗法。
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