Vardas Konstantinos, Ilia Stavroula, Sertedaki Amalia, Charmandari Evangelia, Briassouli Efrossini, Goukos Dimitris, Apostolou Kleovoulos, Psarra Katerina, Botoula Efthimia, Tsagarakis Stylianos, Magira Eleni, Routsi Christina, Stratakis Constantine A, Nanas Serafim, Briassoulis George
First Critical Care Department, National and Kapodistrian University of Athens, Athens, Greece.
Pediatric Intensive Care Unit, University Hospital, University of Crete, 71500, Heraklion, Crete, Greece.
Intensive Care Med Exp. 2017 Dec;5(1):10. doi: 10.1186/s40635-017-0123-8. Epub 2017 Feb 21.
The purposes of this study are to examine if the human glucocorticoid receptor (hGR) isoform-α mRNA and hGR protein expressions are deficient in the acute phase of sepsis (S) compared to systemic inflammatory response syndrome (SIRS) and healthy subjects (H) and to evaluate if the hGRα and hGR alterations are associated with cortisol changes and if they are related to (1) extracellular and intracellular heat shock proteins (HSP) 72 and 90α; (2) ACTH, prolactin, and interleukins (ILs); and (3) outcome.
Patients consecutively admitted to a university hospital intensive care unit (ICU) with S (n = 48) or SIRS (n = 40) were enrolled in the study. Thirty-five H were also included. Total mRNA was isolated from peripheral blood samples and cDNA was prepared. RT-PCR was performed. Intracellular hGR and HSP expression in monocytes and/or neutrophils was evaluated using four-colour flow cytometry. Serum prolactin, ACTH, and cortisol concentrations were also measured. ELISA was used to evaluate serum ILs and extracellular (e) HSPs (eHSP72, eHSP90α).
hGR protein was higher in S compared to H and SIRS; hGRα mRNA was higher in S compared to H (p < 0.05). In sepsis, hGR protein and eHSP72 were higher among non-survivors compared to survivors (p < 0.05). The hGR MFI and hGRα mRNA fold changes were significantly related to each other (r = 0.64, p < 0.001). Monocyte hGR protein expression was positively correlated with extracellular and intracellular HSPs, cortisol, and ILs and negatively to organ dysfunction (p < 0.05). HSPs, hGR, and cortisol were able to discriminate sepsis from SIRS (AUROC > 0.85, p < 0.05). In sepsis, monocyte-hGR protein and eHSP72 were strong predictors of mortality (AUROC > 0.95, p < 0.04).
Acute-phase sepsis is associated with increased hGR expression and cortisol concentrations, possibly implying no need for exogenous steroids. At this stage, hGR is able to predict sepsis and outcome and is related to stress-activated bio-molecules and organ dysfunction.
本研究旨在探讨与全身炎症反应综合征（SIRS）及健康受试者（H）相比，脓毒症（S）急性期人糖皮质激素受体（hGR）亚型-α mRNA及hGR蛋白表达是否存在缺陷，评估hGRα及hGR的改变是否与皮质醇变化相关，以及是否与以下因素有关：（1）细胞外和细胞内热休克蛋白（HSP）72和90α；（2）促肾上腺皮质激素、催乳素及白细胞介素（ILs）；（3）预后。
连续纳入一所大学医院重症监护病房（ICU）的S患者（n = 48）或SIRS患者（n = 40）进行研究。还纳入了35名健康受试者（H）。从外周血样本中分离总mRNA并制备cDNA。进行逆转录聚合酶链反应（RT-PCR）。使用四色流式细胞术评估单核细胞和/或中性粒细胞内的hGR及HSP表达。还测量血清催乳素、促肾上腺皮质激素及皮质醇浓度。采用酶联免疫吸附测定（ELISA）评估血清ILs及细胞外（e）HSPs（eHSP72、eHSP90α）。
与H和SIRS相比，S患者的hGR蛋白水平更高；与H相比，S患者的hGRα mRNA水平更高（p < 0.05）。在脓毒症中，非存活者的hGR蛋白及eHSP72水平高于存活者（p < 0.05）。hGR平均荧光强度（MFI）及hGRα mRNA倍数变化显著相关（r = 0.64，p < 0.001）。单核细胞hGR蛋白表达与细胞外和细胞内HSPs、皮质醇及ILs呈正相关，与器官功能障碍呈负相关（p < 0.05）。HSPs、hGR及皮质醇能够区分脓毒症和SIRS（受试者工作特征曲线下面积（AUROC）> 0.85，p < 0.05）。在脓毒症中，单核细胞-hGR蛋白及eHSP72是死亡率的强预测指标（AUROC > 0.95，p < 0.04）。
急性期脓毒症与hGR表达及皮质醇浓度升高相关，可能意味着无需使用外源性类固醇。在此阶段，hGR能够预测脓毒症及预后，且与应激激活的生物分子及器官功能障碍有关。
