Antisense basic fibroblast growth factor alters the time course of mitogen-activated protein kinase in arterialized vein graft remodeling
Antisense basic fibroblast growth factor alters the time course of mitogen-activated protein kinase in arterialized vein graft remodeling
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DOI：
10.1067/mva.2003.130
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发表时间：
2003-04-01
影响因子：
4.3
通讯作者：
Sumpio, BE
中科院分区：
文献类型：
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作者：
Yamashita, A;Hanna, AK;Sumpio, BE
目的：在植入动脉循环的兔静脉移植物中，新生内膜增生（NIH）在3周时完成。促分裂原活化蛋白激酶（MAPK）家族的蛋白激酶的活化被认为在重塑事件如细胞增殖、分化和迁移中是关键的，如在NIH中发现的。我们以前证明，反义碱性成纤维细胞生长因子（ASbFGF）抑制碱性成纤维细胞生长因子（bFGF）的合成在美国国立卫生研究院的球囊损伤模型。我们检测了ASbFGF对NIH的影响以及MAPK、细胞外信号调节激酶（ERK）1/2、c-Jun N末端蛋白激酶（JNK）和p38激酶激活的时间过程。方法：在75只新西兰白色家兔中进行颈动脉间置静脉旁路移植术。用含有大鼠ASbFGF信使RNA序列的复制缺陷型腺病毒以每毫升1 × 10（10）空斑形成单位转染颈外静脉节段;对照动物仅给予磷酸盐缓冲盐水溶液（PBS）。在30分钟、4天、7天和21天处死家兔（n = 8）。每组4个移植物用福尔马林固定，石蜡包埋，然后用弹性蛋白-胶原和苏木精-伊红染色。另外四个移植物单独冷冻，并提取总蛋白。用Western印迹分析和免疫组织化学测定MAPK、ERK 1/2、JNK和p38的磷酸化。结果：PBS组和ASbFGF组在21天时的新生内膜厚度分别为60.2 ± 2.1和39.4 ± 2.1 μ m（P <0.01）。在对照组和ASbFGF组中，所有3种MAPK均表现出与植入前水平相比的激活。然而，当与PBS组相比时，ASbFGF组在第4天和第7天显示出对所有三种MAPK的大于33%的抑制（P <0.05），但在第21天的任何MAPK活化方面没有显著差异（P > 0.05，所有组）。结果：PBS组和ASbFGF组移植静脉内膜和外膜中均可见活化的MAPK阳性细胞。用ASbFGF处理的移植物表现出降低的MAPK活化和较少的新生内膜增厚。这些结果表明，静脉移植适应动脉循环的过程，以及随后的NIH，可能取决于碱性成纤维细胞生长因子的活性，这是介导的，至少部分地，由MAPK依赖性机制。
Purpose: Neointimal hyperplasia (NIH) is complete by 3 weeks in rabbit vein grafts implanted into the arterial circulation. Activation of the mitogen-activated protein kinase (MAPK) family of protein kinases is thought to be critical in remodeling events such as cellular proliferation, differentiation, and migration, as found in NIH. We previously demonstrated that antisense basic fibroblast growth factor (ASbFGF) inhibited the synthesis of basic fibroblast growth factor (bFGF) in the balloon injury model of NIH. We examined the effect of ASbFGF on NIH and the time course of MAPK, extracellular signal-regulated kinase (ERK) 1/2, c-Jun N-terminal protein kinase (JNK), and p38 kinase activation in arterialized. vein grafts.Methods: Carotid interposition of a vein bypass graft was performed in 75 New Zealand White rabbits. Segments of the external jugular vein were transfected with a replication-deficient adenovirus containing the messenger RNA sequence for rat ASbFGF at 1 x 10(10) plaque-forming units per milliliter; control animals were given phosphate-buffered saline solution (PBS) alone. Rabbits were killed at 30 minutes, 4 days, 7 days, and 21 days (n = 8). Four grafts in each group were fixed with formalin and embedded in paraffin, then processed with elastin-collagen and hematoxylin-eosin stains. The other four grafts were individually frozen, and total protein was extracted. Phosphorylation of MAPK, ERK1/2, JNK, and p38, was determined with Western blot analysis and immunohistochemistry. Groups were compared with analysis of variance.Results: The thickness of neointima in the PBS group and the ASbFGF group at 21 days was 60.2 +/- 2.1 and 39.4 +/- 2.1 mum, respectively (P < .01). In both the control and ASbFGF groups, all 3 MAPKs demonstrated activation compared with preimplantation levels. However, when compared with the PBS group the ASbFGF group showed greater than 33% inhibition of all three MAPKs by day 4 and day 7 (P < .05), but no significant difference in any MAPK activation by day 21 (P > .05, all groups). Cells staining positive for activated MAPK were found in the neointima and adventitia of vein grafts in both the PBS and ASbFGF groups.Conclusion: MAPKs are activated during the first week after vein graft implantation. Grafts treated with ASbFGF demonstrated reduced MAPK activation and less neointimal thickening. These results suggest that the process of vein graft adaptation to the arterial circulation, and subsequent NIH, may depend on basic fibroblast growth factor activity, which is mediated, at least in part, by a MAPK-dependent mechanism.