论文部分内容阅读
选择大鼠主动脉平滑肌细胞作为细胞模型 ,观察丝裂原活化蛋白激酶 ( MAPK)信号通路在血小板源生长因子 ( PDGF)诱导的血管平滑肌细胞( VSMC)增殖中的作用。用 [3 H]Td R参入率作为衡量 VSMC增殖的指标 ,以 Western-印迹方法 ,用磷酸化一抗测 MAPK磷酸化的程度作为衡量 MAPK活性的指标 ,实验结果发现 ,PDGF( 0 .0 2 - 2 0μg·L-1)诱导 VSMC增殖呈剂量依赖性。 PDGF( 2μg· L-1)能持续性激活 MAPK。用 PD980 59( 1 0 -1 0 0 mol· L-1)预处理 1 5min后 ,MAPK活性较对照组均有明显差别 ( P<0 .0 5) .MAPK反义寡核苷酸能明显抑制 PDGF诱导的 MAPK的激活 ,并明显抑制 VSMC[3 H]Td R参入。上述结果表明 ,MAPK参与 PDGF促细胞增殖的信号途经 ,它的持续性激活与细胞增殖有关 ,针对 p42 /p44MAPK设计的反义寡核苷酸类能有效抑制 PDGF诱导的血管平滑肌细胞增殖。
Rat aortic smooth muscle cells were selected as the cell model to observe the role of mitogen - activated protein kinase (MAPK) signaling pathway in the proliferation of vascular endothelial growth factor (PDGF) - induced vascular smooth muscle cells (VSMCs). The [3 H] Td R incorporation was used as a measure of VSMC proliferation. Western blotting was used to measure the phosphorylation of MAPK using phospho-1 antibody as a measure of MAPK activity. The results showed that PDGF (0. 02 - 2 0μg · L-1) induced VSMC proliferation in a dose-dependent manner. PDGF (2μg · L-1) can activate MAPK persistently. After pretreatment with PD980 59 (1 0 -1 0 0 mol · L-1) for 15 min, MAPK activity was significantly lower than that of the control group (P <0.05) .MAPK antisense oligonucleotide significantly inhibited PDGF-induced activation of MAPK, and significantly inhibited VSMC [3 H] Td R incorporation. The above results indicate that MAPK is involved in the signaling pathway of PDGF-induced cell proliferation, and its sustained activation is related to cell proliferation. Antisense oligonucleotides targeting p42 / p44MAPK effectively inhibit PDGF-induced proliferation of vascular smooth muscle cells.