Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China.
Department of Physiology, Medical School of Southeast University, Nanjing, Jiangsu, 210009, China.
Sci Rep. 2018 Jan 29;8(1):1735. doi: 10.1038/s41598-018-20195-6.
Monocyte chemotactic protein-1-induced protein 1 (MCPIP1) plays a important role in ischemia/reperfusion (I/R) injury. Autophagy is involved in activating endothelial cells in response to I/R. However, researchers have not clearly determined whether MCPIP1 mediates I/R injury in endothelial cells via autophagy, and its downstream mechanism remains unclear. Western blotting analyses and immunocytochemistry were applied to detect protein levels were detected in HUVECs. An in vitro scratch assay was used to detect cell migration. Cells were transfected with siRNAs to knockdown MCPIP1 and high mobility group box 1 (HMGB1) expression. The pharmacological activator of autophagy rapamycin and the specific calcium-sensing receptor (CaSR) inhibitor NPS-2143 were used to confirm the roles of autophagy and CaSR in I/R injury. I/R induced HMGB1 and CaSR expression, which subsequently upreguated the migration and apoptosis of HUVECs and coincided with the increase of autophagy. HMGB1 was involved in cell migration, whereas CaSR specifically participated in I/R-induced HUVEC apoptosis. Based on these findings, I/R-induced MCPIP1 expression regulates the migration and apoptosis of HUVECs via HMGB1 and CaSR, respectively, suggesting a new therapeutic targetof I/R injury.
单核细胞趋化蛋白-1 诱导蛋白 1（MCPIP1）在缺血/再灌注（I/R）损伤中发挥重要作用。自噬参与了内皮细胞对 I/R 的反应。然而，研究人员尚未明确确定 MCPIP1 是否通过自噬介导内皮细胞的 I/R 损伤，其下游机制仍不清楚。Western blot 分析和免疫细胞化学用于检测 HUVECs 中的蛋白水平。体外划痕实验用于检测细胞迁移。用 siRNA 转染细胞以敲低 MCPIP1 和高迁移率族蛋白 B1（HMGB1）的表达。自噬的药理学激活剂雷帕霉素和特异性钙敏感受体（CaSR）抑制剂 NPS-2143 用于确认自噬和 CaSR 在 I/R 损伤中的作用。I/R 诱导 HMGB1 和 CaSR 的表达，随后上调 HUVEC 的迁移和凋亡，同时自噬增加。HMGB1 参与细胞迁移，而 CaSR 特异性参与 I/R 诱导的 HUVEC 凋亡。基于这些发现，I/R 诱导的 MCPIP1 表达通过 HMGB1 和 CaSR 分别调节 HUVEC 的迁移和凋亡，提示 I/R 损伤的新治疗靶点。
