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OGR1 mediates the inhibitory effects of acidic environment on proliferation and angiogenesis of endothelial progenitor cells

机译:OGR1介导酸性环境对内皮祖细胞增殖和血管生成的抑制作用

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Abstract Ovarian cancer G‐protein‐coupled receptor 1 (OGR1), an acid‐sensitive receptor, plays a key proton‐sensing role through stimulation of inositol phosphate formation. Avascular necrosis of the femoral head is characterized by apoptosis of bone cells mainly resulting from deficient local blood perfusion, eventually leading to acidification with disruption of endothelial progenitor cells’ (EPCs) function. However, whether EPCs express OGR1 has not been demonstrated. This study attempted to whether OGR1 mediates the effects of acid on proliferation, migration, and angiogenesis in EPCs. FITC‐UEA‐I and Dil‐Ac‐LDL double‐staining methods were used to identify EPCs. Expression of OGR1 was analyzed by RT‐PCR (reverse transcription PCR) and western blot after incubation in media ranging in pH, cell counting kit‐8 and cell cycle analysis were used to analyze proliferation and cell cycle distribution. Scratch test, transwell migration assay, and tube formation experiments were performed to analyze migration and vascularization of EPCs after silencing OGR1 with small interfering RNA (siRNA). The result show EPCs were positive for FITC‐UEA‐I and Dil‐Ac‐LDL double‐staining and expressed OGR1. The expression of OGR1 increased gradually with decreased pH and was highest in pH 6.4 medium. Incubation in pH 6.4 medium inhibited proliferation of EPCs and caused cell cycle arrest. Silencing of OGR1 using siRNA partially reversed the effect of acidic environment on EPCs. Migration and angiogenesis of EPCs were inhibited in pH 6.4 medium, and silencing of OGR1 partially reversed this effect. The results demonstrated expression of OGR1 in EPCs, and the OGR1 mediated the effects of acidic environment on proliferation, migration, and angiogenesis of EPCs.
机译:摘要卵巢癌G蛋白偶联受体1(OGR1),一种酸敏感受体,通过刺激肌醇磷酸盐形成,起到关键的质子传感作用。股骨头的缺血性坏死的特征在于骨细胞的凋亡,主要是由于缺乏局部血液灌注,最终导致酸化具有内皮祖细胞的破坏(EPCS)功能。但是,EPCS表达OGR1是否尚未证明。该研究试图对EGR1是否介导酸对EPC中的增殖,迁移和血管生成的影响。 FITC-UEA-I和DIL-AC-LDL双染料方法用于鉴定EPCS。通过RT-PCR(逆转录PCR)分析OGR1的表达,蛋白质印迹在pH中的培养中孵育后,用于分析细胞计数试剂盒-8和细胞循环分析来分析增殖和细胞周期分布。进行刮擦试验,Transwell迁移测定和管形成实验,以分析沉默于小干扰RNA(siRNA)后沉默的EGCS的迁移和血管化。结果表明EPC对于FITC-UEA-I和DIL-AC-LDL双染色和表达的OGR1是阳性的。 OGR1的表达随着pH降低而逐渐增加,pH 6.4培养基中最高。在pH6.4中孵育培养基抑制EPC的增殖并引起细胞周期骤停。使用siRNA的OGR1沉默部分逆转酸性环境对EPC的影响。在pH6.4培养基中抑制了EPC的迁移和血管生成,并且OGR1的沉默部分逆转了这种效果。结果证明了OGR1在EPC中的表达,OGR1介导酸性环境对EPC的增殖,迁移和血管生成的影响。

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