MULTIPLE GROWTH-FACTORS ARE RELEASED FROM MECHANICALLY INJURED VASCULAR SMOOTH-MUSCLE CELLS
MULTIPLE GROWTH-FACTORS ARE RELEASED FROM MECHANICALLY INJURED VASCULAR SMOOTH-MUSCLE CELLS
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DOI：
10.1152/ajpheart.1995.269.5.h1641
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发表时间：
1995-11-01
影响因子：
4.8
通讯作者：
HORWITZ, LD
中科院分区：
文献类型：
--
作者：
CROWLEY, ST;RAY, CJ;HORWITZ, LD
在血管成形术诱导的血管损伤过程中，促有丝分裂和趋化信号的局部释放可能引发再狭窄。我们调查是否机械损伤血管平滑肌细胞（VSMC）的生物活性肽生长因子的释放的结果。牛平滑肌细胞培养物的单层被细胞刮擦机械损伤。收集来自对照和损伤SMC培养物的条件培养基（CM），并通过受体SMC培养物中[H-3]胸苷掺入来测量促有丝分裂活性。损伤CM的促有丝分裂活性在损伤后15分钟内检测到。损伤后15分钟取出损伤细胞的CM，用无血清培养基替换，损伤后1-6天检测，替换CM无促有丝分裂活性。苏拉明，一种非特异性肽生长因子拮抗剂，显着抑制损伤CM的促有丝分裂活性。通过免疫印迹分析，在来自损伤细胞的CM中检测到碱性成纤维细胞生长因子（bFGF）、血小板衍生生长因子（PDGF A链）和表皮生长因子（EGF）。bFGF（34%）、PDGF-AA（32%）、PDGF-BB（25%）和EGF（25%）的中和抗体可显著抑制受损CM的促有丝分裂活性。转化生长因子（TGF）-β的中和抗体没有影响。总之，bFGF、PDGF和EGF从机械损伤的VSMC中立即释放。VSMC可能含有预先形成的生物活性生长因子，这些生长因子在机械损伤后从细胞质中有效释放。损伤VSMC的条件培养基具有高度的促有丝分裂活性，这种活性可能是由于多种生长因子协同作用的结果。
Local release of mitogenic and chemotactic signals during angioplasty-induced vascular injury may initiate restenosis. We investigated whether mechanical injury to vascular smooth muscle cells (VSMC) results in the release of biologically active peptide growth factors. Monolayers of bovine SMC cultures were mechanically injured by cell scraping. Conditioned medium (CM) from control and injured SMC cultures was collected, and the mitogenic activity was measured by [H-3]thymidine incorporation in recipient SMC cultures. Mitogenic activity from injured CM was detected within 15 min after injury. When the CM from injured cells was removed 15 min after injury and replaced with serum-free media, there was no detectabe mitogenic activity in the replacement CM assessed 1-6 days postinjury. Suramin, a nonspecific peptide growth factor antagonist, significantly inhibited the mitogenic activity of injured CM. Basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF A chain), and epidermal growth factor (EGF) were detected in CM from injured cells by immunoblot analysis. The mitogenic activity of injured CM was significantly inhibited with neutralizing antibodies to bFGF (34%), PDGF-AA (32%), PDGF-BB (25%), and EGF (25%). A neutralizing antibody to tranforming growth factor (TGF)-beta had no effect. In conclusion, bFGF, PDGF, and EGF are immediately released from mechanically injured VSMC. VSMC Likely contain preformed, biologically active growth factors that are efficiently released from the cell cytoplasm following mechanical injury. Conditioned medium from injured VSMC is highly mitogenic, and this activity is probably due to multiple growth factors interacting synergistically.