Gråberg Truls, Strömmer Lovisa, Hedman Erik, Uzunel Mehmet, Ehrenborg Ewa, Wikström Ann-Charlotte
Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Solna, Sweden.
Department of Clinical Pharmacology, Karolinska University Hospital, Solna, Sweden.
J Inflamm Res. 2015 Aug 19;8:149-60. doi: 10.2147/JIR.S84165. eCollection 2015.
An assay to determine glucocorticoid (GC) responsiveness in humans could be used to monitor GC non-responsiveness in states of GC insufficiency and could provide a tool to adapt GC treatment to individual patients. We propose an ex vivo assay to test GC responsiveness in peripheral leukocytes. The assay was evaluated in a human experimental model of surgery-induced inflammation.
Changes in expression of the GC-regulated genes GILZ, IL1R2, FKBP5, and HLA-DR and glucocorticoid receptor alpha (GRα) were determined by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in peripheral leukocytes from surgical patients and healthy blood donors (total n=60) in response to low (1 nM) and high (1 µM) dexamethasone (DEX). The final selection of a suitable endogenous control gene was based on the studies of stability during DEX treatment and inflammation. Correlations between pre- and postoperative GC-induced gene expression, the postoperative systemic inflammatory and metabolic response (CRP, IL-6, white blood cell count, cytokines, resistin, free fatty acids, glucose, insulin, and adiponectin), and the clinical outcome were analyzed. The length of stay in the intensive care unit (ICU-LOS), the length of stay in the hospital, and postoperative complications were used to measure clinical outcome.
When the blood donors were compared to the patients, there were no significant differences in the regulation of the genes in response to DEX, except for GRα. Preoperative, but not postoperative, gene regulation of GILZ and GRα was negatively correlated to ICU-LOS (P<0.05 and P<0.01, respectively). Preoperative GILZ and FKBP5 gene regulation was negatively correlated to postoperative systemic TNFα and MIP-1α levels.
We suggest that this assay could be used to determine GC responsiveness. An alteration in preoperative GC responsiveness may be related to a patient's ability to recover from surgically induced inflammatory stress.
一种用于测定人类糖皮质激素（GC）反应性的检测方法可用于监测GC不足状态下的GC无反应性，并可为根据个体患者调整GC治疗提供一种工具。我们提出了一种体外检测方法来测试外周血白细胞中的GC反应性。该检测方法在手术诱导炎症的人体实验模型中进行了评估。
通过逆转录定量聚合酶链反应（RT-qPCR）测定手术患者和健康献血者（共60例）外周血白细胞中GC调节基因GILZ、IL1R2、FKBP5和HLA-DR以及糖皮质激素受体α（GRα）的表达变化，以响应低剂量（1 nM）和高剂量（1 µM）地塞米松（DEX）。合适内参基因的最终选择基于DEX治疗和炎症期间稳定性的研究。分析术前和术后GC诱导基因表达、术后全身炎症和代谢反应（CRP、IL-6、白细胞计数、细胞因子、抵抗素、游离脂肪酸、葡萄糖、胰岛素和脂联素）与临床结局之间的相关性。重症监护病房住院时间（ICU-LOS）、住院时间和术后并发症用于衡量临床结局。
将献血者与患者进行比较时，除GRα外，基因对DEX反应的调节无显著差异。术前而非术后，GILZ和GRα的基因调节与ICU-LOS呈负相关（分别为P<0.05和P<0.01）。术前GILZ和FKBP5基因调节与术后全身TNFα和MIP-1α水平呈负相关。
我们认为该检测方法可用于测定GC反应性。术前GC反应性的改变可能与患者从手术诱导的炎症应激中恢复的能力有关。
