Chen Hai-Feng, Xie Liang-Di, Xu Chang-Sheng
Fujian Hypertension Research Institute, First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China.
Mol Cell Biochem. 2009 Jul;327(1-2):1-6. doi: 10.1007/s11010-009-0034-4. Epub 2009 Feb 4.
The aim of the present study was to investigate the role of heat shock protein 27 (HSP27) phosphorylation in the migration of vascular smooth muscle cells (VSMCs) induced by angiotensin II (AngII) and platelet derived growth factor-BB (PDGF-BB).
The activity of HSP27 was evaluated by Western blot with specific phospho-HSP27 antibody. F-actin polymerization was detected by FITC-Phalloidine staining using confocal microscopy. Modified Boyden chamber technique was employed for VSMCs migration assessment.
The phosphorylation of HSP27 was induced by AngII and PDGF-BB in a time- and concentration-dependent manner in VSMCs, which was significantly blocked by the HSP inhibitor Quercetin in a concentration-dependent manner. Reorganization of actin stimulated by AngII and PDGF-BB was markedly inhibited by pretreatment with 100 micromol/l Quercetin. The migration of VSMCs induced by AngII and PDGF-BB was partially inhibited by Quercetin with peak inhibition concentration at 100 micromol/l.
HSP27 phosphorylation plays an important role in mediating the rearrangement of F-actin and migration of VSMCs induced by AngII and PDGF-BB. HSP27 may be a potential target for the interventional treatment of pathological process related to cell migration.
本研究旨在探讨热休克蛋白27（HSP27）磷酸化在血管紧张素II（AngII）和血小板衍生生长因子-BB（PDGF-BB）诱导的血管平滑肌细胞（VSMC）迁移中的作用。
使用特异性磷酸化HSP27抗体通过蛋白质印迹法评估HSP27的活性。使用共聚焦显微镜通过FITC-鬼笔环肽染色检测F-肌动蛋白聚合。采用改良的Boyden小室技术评估VSMC迁移。
AngII和PDGF-BB在VSMC中以时间和浓度依赖性方式诱导HSP27磷酸化，HSP抑制剂槲皮素以浓度依赖性方式显著阻断该磷酸化。用100微摩尔/升槲皮素预处理可显著抑制AngII和PDGF-BB刺激的肌动蛋白重组。AngII和PDGF-BB诱导的VSMC迁移被槲皮素部分抑制，最大抑制浓度为100微摩尔/升。
HSP27磷酸化在介导AngII和PDGF-BB诱导的F-肌动蛋白重排和VSMC迁移中起重要作用。HSP27可能是与细胞迁移相关病理过程介入治疗的潜在靶点。
