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目的研究替米沙坦对内皮祖细胞增殖、迁移、黏附等生物学活性的影响并探讨其可能机制。方法利用密度梯度离心法分离、培养人外周血单个核细胞,经FITC-UEA-I和Dil-acLDL双染色鉴定为正在分化的内皮祖细胞。将分离、培养的内皮祖细胞分为对照组、替米沙坦组(0.1μmol/L、1μmol/L、10μmol/L)、过氧化体增殖物激活型受体γ抑制剂(GW9662)干预组和磷脂酰肌醇-3-羟基激酶抑制剂(Ly294002)干预组。采用MTT比色法、Transwell小室、细胞计数法观察各组内皮祖细胞增殖、迁移、黏附能力的变化情况。同时采用免疫蛋白印迹法观察替米沙坦处理内皮祖细胞后丝苏氨酸蛋白激酶及磷酸化丝苏氨酸蛋白激酶的表达情况。结果与对照组相比,替米沙坦组内皮祖细胞增殖、迁移、黏附能力显著提高,且在不同浓度组间呈剂量依赖性增强,而在过氧化体增殖物激活型受体γ抑制剂干预组和磷脂酰肌醇-3-羟基激酶抑制剂干预组,内皮祖细胞功能活性的改善受到明显的抑制。免疫蛋白印迹检测显示,相比于对照组,替米沙坦组磷酸化丝苏氨酸蛋白激酶的表达水平明显增高,而在过氧化体增殖物激活型受体γ抑制剂干预组磷酸化丝苏氨酸蛋白激酶的表达相比于对照组未见明显增高。结论替米沙坦具有促进内皮祖细胞增殖、迁移、黏附等功能的作用,其主要机制可能与过氧化体增殖物激活型受体γ介导的磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶信号通路激活有关。
Objective To study the effects of telmisartan on the biological activity of endothelial progenitor cells (EPCs) such as proliferation, migration and adhesion and to explore its possible mechanism. Methods Human peripheral blood mononuclear cells were isolated and cultured by density gradient centrifugation. The differentiated endothelial progenitor cells were identified by double staining with FITC-UEA-I and Dil-acLDL. The cultured EPCs were divided into control group, telmisartan group (0.1μmol / L, 1μmol / L, 10μmol / L) and peroxisome proliferator activated receptor γ inhibitor (GW9662) And phosphatidylinositol 3-hydroxy kinase inhibitor (Ly294002) intervention group. The changes of proliferation, migration and adhesion of endothelial progenitor cells were observed by MTT assay, Transwell chamber and cell counting method. Western blotting was used to observe the expression of serine protein kinase and phospho-threonine protein kinase after treatment with telmisartan. Results Compared with the control group, the EPCs proliferation, migration and adhesion were significantly increased in the telmisartan group and increased in a dose-dependent manner in different concentration groups. However, in the group of telmisartan, Intervention group and phosphatidylinositol 3-hydroxy kinase inhibitor intervention group, the improvement of endothelial progenitor activity was significantly inhibited. Immunoblotting showed that compared with the control group, the expression of phosphorylated threonine protein kinase was significantly increased in the telmisartan group, whereas phosphorylated silk fibroin in the intervention group of the inhibitor of peroxisome proliferator-activated receptor γ Threonine protein kinase expression compared to the control group no significant increase. Conclusion Telmisartan can promote the proliferation, migration and adhesion of endothelial progenitor cells. The main mechanism may be related to the activation of phosphatidylinositol 3-kinase / threonine by peroxisome proliferator-activated receptor γ Acid protein kinase signaling pathway activation.