Department of Anesthesia, Kyoto University Hospital, 54 Shogoin-Kawaracho, Sakyo-Ku, Kyoto, 6060-8507, Japan.
J Anesth. 2010 Feb;24(1):54-60. doi: 10.1007/s00540-009-0829-1. Epub 2009 Dec 29.
Hypoxia-inducible factor 1 (HIF-1) is a master transcription factor of hypoxia-induced gene expression. Anesthetics and perioperative drugs have been reported to affect HIF-1 activity. However, the effect of propofol on HIF-1 activity is not well documented. In this study, we investigated the effect of propofol on HIF-1 activation using macrophage-differentiated THP-1 cells.
Cells were exposed to lipopolysaccharide (LPS) under 20 or 1% O(2) conditions with or without propofol treatment. The cell lysate was subjected to Western blot analysis using anti-HIF-1alpha and HIF-1beta antibodies. HIF-1-dependent gene expression was investigated by quantitative real-time reverse-transcriptase PCR analysis and luciferase assay. The amount of cellular lactate and ATP was assayed.
Propofol suppressed HIF-1alpha protein accumulation induced by LPS, but not by hypoxia in the THP-1 cells in a dose-dependent manner by inhibiting the neo-synthesis of HIF-1alpha protein. Induction of the HIF-1 downstream gene expression including glucose transporter 1, enolase 1, lactate dehydrogenase A, pyruvate dehydrogenase kinase-1 and vascular endothelial growth factor was inhibited by propofol. Propofol suppressed LPS-induced lactate accumulation and ATP content in THP-1 cells.
Our experimental results indicate that propofol inhibits HIF-1 activation and downstream gene expression induced by LPS and suppressed HIF-1-dependent glucose metabolic reprogramming. HIF-1 suppression by propofol in macrophages may explain molecular mechanisms behind the inhibitory effect of propofol on cellular inflammatory responses.
缺氧诱导因子 1（HIF-1）是缺氧诱导基因表达的主要转录因子。有报道称，麻醉剂和围手术期药物会影响 HIF-1 活性。然而，丙泊酚对 HIF-1 活性的影响尚未得到充分证实。在这项研究中，我们使用巨噬细胞分化的 THP-1 细胞研究了丙泊酚对 HIF-1 激活的影响。
细胞在 20%或 1%O(2)条件下用或不用丙泊酚处理，暴露于脂多糖（LPS）下。用抗 HIF-1alpha 和 HIF-1beta 抗体对细胞裂解物进行 Western blot 分析。通过实时定量逆转录 PCR 分析和荧光素酶测定研究 HIF-1 依赖性基因表达。测定细胞内乳酸和 ATP 的量。
丙泊酚以剂量依赖的方式抑制 LPS 诱导的 THP-1 细胞中 HIF-1alpha 蛋白的积累，但不抑制缺氧诱导的 HIF-1alpha 蛋白的新合成，从而抑制 HIF-1alpha 蛋白的积累。诱导包括葡萄糖转运蛋白 1、烯醇酶 1、乳酸脱氢酶 A、丙酮酸脱氢酶激酶-1 和血管内皮生长因子在内的 HIF-1 下游基因表达被丙泊酚抑制。丙泊酚抑制 LPS 诱导的 THP-1 细胞中乳酸积累和 ATP 含量。
我们的实验结果表明，丙泊酚抑制 LPS 诱导的 HIF-1 激活和下游基因表达，并抑制 HIF-1 依赖性葡萄糖代谢重编程。丙泊酚在巨噬细胞中抑制 HIF-1 可能解释了丙泊酚抑制细胞炎症反应的分子机制。
