Loop Torsten, Scheiermann Patrick, Doviakue David, Musshoff Frank, Humar Matjaz, Roesslein Martin, Hoetzel Alexander, Schmidt Rene, Madea Burkhard, Geiger Klaus K, Pahl Heike L, Pannen Benedikt H J
Department of Anesthesiology, University Hospital, Freiburg, Germany.
Anesthesiology. 2004 Sep;101(3):710-21. doi: 10.1097/00000542-200409000-00020.
Modulation of immune defense mechanisms by volatile anesthetics during general anesthesia may compromise postoperative immune competence and healing reactions and affect the infection rate and the rate of tumor metastases disseminated during surgery. Several mechanisms have been suggested to account for these effects. The current study was undertaken to examine the molecular mechanisms underlying these observations.
Effects of sevoflurane, isoflurane, and desflurane were studied in vitro in primary human CD3 T-lymphocytes. DNA-binding activity of the transcription factor activator protein-1 (AP-1) was assessed using an electrophoretic mobility shift assay. Phorbol-myristate-acetate-dependent effects of sevoflurane on the phosphorylation of the mitogen-activated protein kinases were studied using Western blots, the trans-activating potency of AP-1 was determined using reporter gene assays, and the cytokine release was measured using enzyme-linked immunosorbent assays.
Sevoflurane inhibited activation of the transcription factor AP-1. This effect was specific, as the activity of nuclear factor kappabeta, nuclear factor of activated T cells, and specific protein-1 was not altered and several other volatile anesthetics studied did not affect AP-1 activation. Sevoflurane-mediated suppression of AP-1 could be observed in primary CD3 lymphocytes from healthy volunteers, was time-dependent and concentration-dependent, and occurred at concentrations that are clinically achieved. It resulted in an inhibition of AP-1-driven reporter gene activity and of the expression of the AP-1 target gene interleukin-3. Suppression of AP-1 was associated with altered phosphorylation of p38 mitogen-activated protein kinases.
The data demonstrate that sevoflurane is a specific inhibitor of AP-1 and may thus provide a molecular mechanism for the antiinflammatory effects associated with sevoflurane administration.
全身麻醉期间挥发性麻醉药对免疫防御机制的调节可能会损害术后免疫能力和愈合反应，并影响感染率以及手术期间肿瘤转移的发生率。已经提出了几种机制来解释这些影响。本研究旨在探讨这些观察结果背后的分子机制。
在原代人CD3 T淋巴细胞中体外研究七氟醚、异氟醚和地氟醚的作用。使用电泳迁移率变动分析评估转录因子激活蛋白-1（AP-1）的DNA结合活性。使用蛋白质免疫印迹法研究七氟醚对佛波酯-肉豆蔻酸酯-乙酸酯依赖性丝裂原活化蛋白激酶磷酸化的影响，使用报告基因分析确定AP-1的反式激活能力，并使用酶联免疫吸附测定法测量细胞因子释放。
七氟醚抑制转录因子AP-1的激活。这种作用具有特异性，因为核因子κB、活化T细胞核因子和特异性蛋白-1的活性未改变，并且所研究的其他几种挥发性麻醉药不影响AP-1的激活。在健康志愿者的原代CD3淋巴细胞中可观察到七氟醚介导的AP-1抑制，呈时间依赖性和浓度依赖性，且发生在临床可达到的浓度。它导致AP-1驱动的报告基因活性和AP-1靶基因白细胞介素-3的表达受到抑制。AP-1的抑制与p38丝裂原活化蛋白激酶磷酸化的改变有关。
数据表明七氟醚是AP-1的特异性抑制剂，因此可能为与七氟醚给药相关的抗炎作用提供分子机制。
