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目的:细胞基质蛋白FRP(Follistatin related protein,FRP)具有调节细胞增殖与迁移的功能,探讨诱导体外培养血管平滑肌细胞表达FRP的机制研究。方法:分离培养脐动脉和人乳内动脉血管平滑肌细胞(vascularsmooth muscle cell,VSMC);确定植物雌激素类药物大豆苷元抑制VSMC增殖的最佳作用时间与浓度;鉴定雌激素受体α、β;给予大豆苷元、17-β雌二醇和雌激素受体完全阻断剂ICI182780刺激细胞,进行竞争性反转录PCR、Northern Blot和原位杂交探讨药物诱导FRP表达途径。结果:大豆苷元可抑制两种VSMC的增殖;脐动脉VSMC表达雌激素受体α,人乳内动脉VSMC表达受体α、β;大豆苷元和雌激素可显著诱导FRP在两种不同VSMC中的表达,ICI182780可抑制大豆苷元的诱导作用。结论:细胞基质蛋白FRP可被雌激素类药物通过其受体诱导。
OBJECTIVE: FRP (Follistatin related protein, FRP) has functions of regulating cell proliferation and migration, and explores the mechanism of inducing FRP expression in vascular smooth muscle cells. METHODS: Umbilical artery and human intramural arterial smooth muscle cells (VSMCs) were isolated and cultured. The optimal time and concentration of phytoestrogen daidzein to inhibit the proliferation of VSMC were determined. The estrogen receptors α and β were identified. Administration of daidzein, 17-beta estradiol and estrogen receptor complete blocker ICI182780 to stimulate cells, competitive reverse transcription PCR, Northern Blot and in situ hybridization to explore drug-induced FRP expression pathway. RESULTS: Daidzein could inhibit the proliferation of two kinds of VSMCs; umbilical artery VSMC expresses estrogen receptor α, human mammary artery VSMC expresses receptors α and β; daidzein and estrogen can significantly induce FRP in two different VSMCs. In the expression, ICI182780 can inhibit the induction of daidzein. CONCLUSION: The cellular matrix protein FRP can be induced by estrogen drugs through its receptor.