摘要
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。
Aim To investigate the functional effects of telmisartant on endothelial progenitor cells （EPC） func- tional activities. Methods Peripheral blood derived mononuclear cells containing EPC were isolated from healthy vol- unteers and then cultured on fibronectin-coated dishes with endothelial cell growth medium-2 （EBM-2）. The ceils were cultured alone （control groups） ,with telmisartan （0. 1 μmol/L, 1 μmol/L, 10 μmol/L）, or telmisartan plus peroxisome proliferator-activated receptor gamma （PPAR.,/） inhibitor（GW9662） or telmisartan plus PI3K-inhibitor（Ly294002）. The proliferation, migration and adhesion activities of EPC were determined with MTT assay, transwell assay and adhesive as- say, respectively. The expression of Akt and p-Akt were measured by Western Blot analysis. Results In the pres- ence of telmisartan, numbers of colonies increased in a dose-dependent manner. DiI-ac-LDL uptake and lectin staining revealed that these proliferation colonies were EPC. The proliferative, migratory and adhesive activities of EPC were sig- nificantly enhanced after treated with telmisartan in a dose-dependent manner. The inhibition of PPARr and Akt activation attenuated the effect of telmisartan on EPC functions. Meanwhile, the expression of p-Akt were significantly upregu- lated by the treatment of telmisartan. Conclusions Telmisartan could improve the proliferative, migratory and adhe- sive activities of EPC via the PI3K/Akt pathway.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2012年第12期1083-1087,共5页
Chinese Journal of Arteriosclerosis
基金
湖北省教育厅优秀中青年人才项目[Q20112101]
