Fujian Hypertension Research Institute, First Affiliated Hospital, Fujian Medical University, 350005, Fuzhou, China.
Mol Cell Biochem. 2010 Jan;333(1-2):49-56. doi: 10.1007/s11010-009-0203-5. Epub 2009 Jul 17.
The objective of this study is to investigate the signal transduction pathways that regulate heat shock protein 27 (HSP27) phosphorylation and migration of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) 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. Within a given concentration, the phosphorylation of HSP27 induced by AngII and PDGF-BB was blocked by the specific P38MAPK inhibitor SB202190, the specific PI3K inhibitor LY294002 and the specific ERK1/2 inhibitor U0126 in a concentration-dependent manner, with a peak inhibition rate at 87.2%, 78.4% and 37.3%, respectively, induced by AngII (P < 0.01), with a peak inhibition rate at 85.0%, 55.3% and 41.0%, respectively, induced by PDGF-BB (P < 0.01).The migration of VSMCs induced by AngII and PDGF-BB was inhibited by 100 micromol/l SB202190, 30 micromol/l LY294002, and 30 micromol/l U0126, with a inhibition rate at 60.1%, 71.7% and 47.3%, respectively, provoked by AngII (P < 0.01), with a inhibition rate at 55.3%, 55.6% and 38.1%, respectively, provoked by PDGF-BB (P < 0.01). P38MAPK and PI3 K/Akt are important pathways that contribute to the phosphorylation of HSP27 and migration of VSMCs in response to AngII and PDGF-BB. ERK1/2 might be involved in HSP27 phosphorylation and migration of VSMCs provoked by AngII and PDGF-BB.
本研究旨在探讨血管平滑肌细胞（VSMCs）中热休克蛋白 27（HSP27）磷酸化和迁移的信号转导途径，这些途径是由血管紧张素 II（AngII）和血小板衍生生长因子-BB（PDGF-BB）诱导的自发性高血压大鼠（SHR）引起的。通过 Western blot 用特异性磷酸化 HSP27 抗体评估 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 Boyden 室技术评估 VSMCs 的迁移。在一定浓度范围内，特异性 P38MAPK 抑制剂 SB202190、特异性 PI3K 抑制剂 LY294002 和特异性 ERK1/2 抑制剂 U0126 以浓度依赖性方式阻断 AngII 和 PDGF-BB 诱导的 HSP27 磷酸化，其抑制率分别为 AngII 诱导的 87.2%、78.4%和 37.3%（P<0.01），PDGF-BB 诱导的 85.0%、55.3%和 41.0%（P<0.01）。SB202190（100 μmol/L）、LY294002（30 μmol/L）和 U0126（30 μmol/L）可抑制 AngII 和 PDGF-BB 诱导的 VSMCs 迁移，抑制率分别为 AngII 诱导的 60.1%、71.7%和 47.3%（P<0.01），PDGF-BB 诱导的 55.3%、55.6%和 38.1%（P<0.01）。P38MAPK 和 PI3K/Akt 是 HSP27 磷酸化和 AngII 和 PDGF-BB 诱导的 VSMCs 迁移的重要途径。ERK1/2 可能参与 AngII 和 PDGF-BB 诱导的 HSP27 磷酸化和 VSMCs 的迁移。
