Sonna Larry A, Wenger C Bruce, Flinn Scott, Sheldon Holly K, Sawka Michael N, Lilly Craig M
Thermal and Mountain Medicine Div., United States Army Research Institute of Environmental Medicine, Natick, MA 01760, USA.
J Appl Physiol (1985). 2004 May;96(5):1943-53. doi: 10.1152/japplphysiol.00886.2003. Epub 2004 Feb 20.
This study examined gene expression changes associated with exertional heat injury (EHI) in vivo and compared these changes to in vitro heat shock responses previously reported by our laboratory. Peripheral blood mononuclear cell (PBMC) RNA was obtained from four male Marine recruits (ages 17-19 yr) who presented with symptoms consistent with EHI, core temperatures ranging from 39.3 to 42.5 degrees C, and elevations in serum enzymes such as creatine kinase. Controls were age- and gender-matched Marines from whom samples were obtained before and several days after an intense field-training exercise in the heat ("The Crucible"). Expression analysis was performed on Affymetrix arrays (containing approximately 12,600 sequences) from pooled samples obtained at three times for EHI group (at presentation, 2-3 h after cooling, and 1-2 days later) and compared with control values (average signals from two chips representing pre- and post-Crucible samples). After post hoc filtering, the analysis identified 361 transcripts that had twofold or greater increases in expression at one or more of the time points assayed and 331 transcripts that had twofold or greater decreases in expression. The affected transcripts included sequences previously shown to be heat-shock responsive in PBMCs in vitro (including both heat shock proteins and non-heat shock proteins), a number of sequences whose changes in expression had not previously been noted as a result of in vitro heat shock in PBMCs (including several interferon-induced sequences), and several nonspecific stress response genes (including ubiquitin C and dual-specificity phosphatase-1). We conclude that EHI produces a broad stress response that is detectable in PBMCs and that heat stress per se can only account for some of the observed changes in transcript expression. The molecular evidence from these patients is thus consistent with the hypothesis that EHI can result from cumulative effects of multiple adverse interacting stimuli.
本研究检测了与运动性热损伤（EHI）相关的体内基因表达变化，并将这些变化与我们实验室之前报道的体外热休克反应进行了比较。外周血单个核细胞（PBMC）RNA取自四名出现与EHI一致症状、核心体温在39.3至42.5摄氏度之间且血清酶如肌酸激酶升高的男性海军新兵（年龄17 - 19岁）。对照组为年龄和性别匹配的海军陆战队队员，在热环境下进行高强度野外训练演习（“严峻考验”）之前和之后数天采集样本。对EHI组在三个时间点（就诊时、降温后2 - 3小时以及1 - 2天后）采集的混合样本在Affymetrix阵列（包含约12,600个序列）上进行表达分析，并与对照值（代表演习前和演习后样本的两个芯片的平均信号）进行比较。经过事后筛选，分析确定了361个转录本在一个或多个检测时间点表达增加两倍或更多，以及331个转录本表达减少两倍或更多。受影响的转录本包括先前在体外PBMC中显示对热休克有反应的序列（包括热休克蛋白和非热休克蛋白）、一些其表达变化先前未因PBMC体外热休克而被注意到的序列（包括几个干扰素诱导序列）以及几个非特异性应激反应基因（包括泛素C和双特异性磷酸酶 - 1）。我们得出结论，EHI会产生一种在PBMC中可检测到的广泛应激反应，并且热应激本身只能解释部分观察到的转录本表达变化。因此，这些患者的分子证据与EHI可能由多种不良相互作用刺激的累积效应导致这一假设一致。
