首页> 外文期刊>American Journal of Physiology >Stretch activates heparin-binding EGF-like growth factor expression in bladder smooth muscle cells.
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Stretch activates heparin-binding EGF-like growth factor expression in bladder smooth muscle cells.

机译:拉伸激活了膀胱平滑肌细胞中肝素结合型EGF样生长因子的表达。

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摘要

Cultured rat bladder smooth muscle cells (SMC) were grown on collagen-coated silicone membranes and subjected to continuous cycles of stretch-relaxation. Semiquantitative RT-PCR analysis revealed a time-dependent increase in heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) mRNA levels after stretch, with maximal levels appearing after 4 h. Immunostaining for proHB-EGF revealed higher levels of HB-EGF protein in the stretched than in the nonstretched SMC. The ANG II receptor type 1 antagonist losartan markedly suppressed stretch-activated HB-EGF expression. ANG II levels were 3.3-fold higher in the stretch- than in the non-stretch-conditioned media. Stretch stimulation of bladder SMC that had been transiently transfected with an HB-EGF promoter-luciferase expression construct resulted in an 11-fold increase in reporter activity. Mechanical stretch induced a 4.7-fold increase in tritiated thymidine incorporation rate, and this was reduced by 25% in the presence of losartan. We conclude that mechanical stretch activates HB-EGF gene expression in bladder SMC and that this is mediated in part by autocrine ANG II secretion.
机译:培养的大鼠膀胱平滑肌细胞(SMC)在胶原蛋白涂层的有机硅膜上生长,并进行连续的舒张松弛循环。半定量RT-PCR分析显示,拉伸后肝素结合表皮生长因子(EGF)样生长因子(HB-EGF)mRNA水平随时间增加,最大水平出现在4 h后。对proHB-EGF的免疫染色显示,与未拉伸的SMC相比,拉伸后的HB-EGF蛋白水平更高。 ANG II受体1型拮抗剂洛沙坦显着抑制了拉伸激活的HB-EGF表达。在拉伸条件下,ANG II水平比在无拉伸条件的培养基中高3.3倍。已经用HB-EGF启动子-荧光素酶表达构建体瞬时转染的膀胱SMC的拉伸刺激导致报告基因活性增加11倍。机械拉伸导致tri化胸苷掺入率增加了4.7倍,在氯沙坦存在下降低了25%。我们得出结论,机械牵张激活了膀胱SMC中的HB-EGF基因表达,并且这部分是由自分泌ANG II分泌介导的。

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