Genistein inhibits pressure-induced expression of c-fos in isolated mesenteric arteries.
Genistein inhibits pressure-induced expression of c-fos in isolated mesenteric arteries.
复制标题
金雀异黄素抑制离体肠系膜动脉中压力诱导的 c-fos 表达。
DOI：
10.1161/01.hyp.34.1.132
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Prewitt,RL
中科院分区：
文献类型：
--
作者：
Miriel,VA;Allen,SP;Schriver,SD;Prewitt,RL
-我们之前已经证明，将分离的肠系膜动脉的腔内压力从90毫米汞柱提高到140毫米汞柱会增加原癌基因的表达。这些原癌基因编码核转录因子，在细胞周期的不同阶段调节目标基因的表达。因此，压力诱导的原癌基因表达可能代表了压力诱导血管平滑肌生长和/或增殖的机制。这项研究的目的是确定与压力诱导的c-fos表达增加有关的细胞内信号。从雄性Wistar大鼠分离肠系膜小动脉，并将其移植到双管室。动脉插管，缓慢平衡到初始状态(90 mm Hg，37℃)，同时用HEPES-重碳酸盐缓冲的Krebs‘s液持续灌流。平衡期结束后，1支动脉腔内压升至140 mm Hg，持续1h。在旨在确定压力诱导的c-fosmRNA表达增加所涉及的细胞内信号的实验中，在压力增加之前，向两个动脉的超融合液库中引入了特定的抑制剂。然后将动脉固定在磷酸盐缓冲福尔马林中，并将其包埋在石蜡块中。将石蜡包埋动脉切片固定于玻片上，用35S标记的核糖核酸探针进行原位杂交，检测c-fos的表达。尼群地平(10μm ol/L)、含EGTA(1~2μ/L)、降钙素C(0.1μm o l/L)或细胞松驰素D(0.4μm ol/L)的无钙Krebs液不能抑制压力诱导的c-fos的表达，但可被染料木素(30 m ol/L)抑制。结果表明，压力诱导的c-fos表达需要酪氨酸激酶的激活，但信号通路不需要细胞外钙进入、完整的肌动蛋白细丝或蛋白激酶C。正如我们先前所表明的，c-fos的表达与壁应力相关。
—We have previously demonstrated that elevating intraluminal pressure from 90 to 140 mm Hg in isolated mesenteric arteries increases the expression of proto-oncogenes. These proto-oncogenes encode nuclear transcription factors that regulate the expression of target genes during various stages of the cell cycle. Thus, pressure-induced proto-oncogene expression may represent a mechanism by which pressure can induce growth and/or proliferation of vascular smooth muscle. The purpose of this study was to determine the intracellular signals that contribute to the pressure-induced increase in c-fosexpression. Small mesenteric arteries were isolated from male Wistar rats and transferred to a dual-vessel chamber. The arteries were cannulated and slowly equilibrated to initial conditions (90 mm Hg, 37°C) while being continuously superfused with a HEPES-bicarbonate–buffered Krebs’ solution. After the equilibration period, the intraluminal pressure in 1 artery was increased to 140 mm Hg for 1 hour. In experiments designed to determine the intracellular signals involved in the pressure-induced increase in c-fosexpression, specific inhibitors were introduced to the superfusate reservoir of both arteries before the pressure increase. The arteries were then fixed in phosphate-buffered formalin and embedded in paraffin blocks. Sections of paraffin-embedded arteries were fixed on slides, and the expression of c-foswas determined by in situ hybridization with the use of35S-labeled riboprobes. The pressure-induced expression of c-foswas not inhibited by nitrendipine (10 μmol/L), a calcium-free Krebs’ solution containing EGTA (1 to 2 mmol/L), calphostin C (0.1 μmol/L), or cytochalasin D (0.4 μmol/L) but was inhibited by genistein (30 μmol/L). The results suggest that activation of a tyrosine kinase is required for pressure-induced c-fosexpression, but the signaling pathway does not require extracellular calcium entry, intact actin filaments, or protein kinase C. As we have shown previously, the expression of c-foscorrelated with wall stress.