Lee Joomin, Ahn Eunsook, Park Wyun Kon, Park Seyeon
Department of Food and Nutrition, Chosun University, Gwangju 61452, Korea.
Department of Applied Chemistry, Dongduk Women's University, Seoul 02748, Korea.
PLoS One. 2016 Sep 9;11(9):e0162214. doi: 10.1371/journal.pone.0162214. eCollection 2016.
Inhalation anesthetics are used to decrease the spinal cord transmission of painful stimuli. However, the molecular or biochemical processes within cells that regulate anesthetic-induced responses at the cellular level are largely unknown. Here, we report the phosphoproteome profile of SH-SY5y human neuroblastoma cells treated with sevoflurane, a clinically used anesthetic. Phosphoproteins were isolated from cell lysates and analyzed using two-dimensional gel electrophoresis. The phosphorylation of putative anesthetic-responsive marker proteins was validated using western blot analysis in cells treated with both sevoflurane and isoflurane. A total of 25 phosphoproteins were identified as differentially phosphorylated proteins. These included key regulators that signal cytoskeletal remodeling steps in pathways related to vesicle trafficking, axonal growth, and cell migration. These proteins included the Rho GTPase, Ras-GAP SH3 binding protein, Rho GTPase activating protein, actin-related protein, and actin. Sevoflurane and isoflurane also resulted in the dissolution of F-actin fibers in SH-SY5y cells. Our results show that anesthetics affect the phosphorylation of proteins involved in cytoskeletal remodeling pathways.
吸入性麻醉剂用于减少脊髓对疼痛刺激的传导。然而，在细胞水平上调节麻醉诱导反应的分子或生化过程在很大程度上尚不清楚。在此，我们报告了用临床使用的麻醉剂七氟醚处理的SH-SY5y人神经母细胞瘤细胞的磷酸化蛋白质组图谱。从细胞裂解物中分离磷酸化蛋白质，并使用二维凝胶电泳进行分析。在用七氟醚和异氟醚处理的细胞中，使用蛋白质印迹分析验证了假定的麻醉反应性标记蛋白的磷酸化。总共25种磷酸化蛋白质被鉴定为差异磷酸化蛋白质。这些包括在与囊泡运输、轴突生长和细胞迁移相关的途径中标记细胞骨架重塑步骤的关键调节因子。这些蛋白质包括Rho GTP酶、Ras-GAP SH3结合蛋白、Rho GTP酶激活蛋白、肌动蛋白相关蛋白和肌动蛋白。七氟醚和异氟醚还导致SH-SY5y细胞中F-肌动蛋白纤维溶解。我们的结果表明，麻醉剂会影响参与细胞骨架重塑途径的蛋白质的磷酸化。
