Metformin Inhibits the Expression of Biomarkers of Fibrosis of EPCs In Vitro
Metformin Inhibits the Expression of Biomarkers of Fibrosis of EPCs In Vitro
复制标题
二甲双胍体外抑制EPCs纤维化生物标志物的表达
DOI：
10.1155/2019/9019648
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发表时间：
2019-01-01
影响因子：
4.3
通讯作者：
Luo, Fanyan
中科院分区：
文献类型：
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作者：
Han, Fei;Shu, Jie;Luo, Fanyan
内皮祖细胞（Endothelial progenitor cells，EPCs）是一类在血管修复和心血管疾病治疗中具有重要功能的循环细胞。然而，在患有心房颤动（AF）的患者中，据报道EPC的数量和功能降低。TGF-β在AF患者中高度表达。在这项研究中，我们研究了TGF-β1对EPCs的作用以及二甲双胍对TGF-β1诱导的EPCs的治疗结果。用不同浓度（5 ng/ml、10 ng/ml和20 ng/ml）的TGF-β1诱导EPCs 48 h，然后进行蛋白质印迹、qPCR和免疫荧光分析，以研究纤维化相关蛋白α-SMA、Col I、Col III、CTGF和MMP-1水平的变化。活-死细胞染色用于评估细胞凋亡。与对照组相比，TGF-β1处理显著（p < 0.05）增加α-SMA、Col I、Col III、CTGF和MMP-1的水平，并呈剂量依赖性。然后使用最有效浓度的TGF-β1（20 ng/ml）诱导EPC中的纤维化生物标志物表达，然后用不同浓度（0.5、1和2 mmol/l）的二甲双胍处理。二甲双胍处理抑制TGF-β诱导的所有上述因子的表达，其中2 mmol/l的效果是显著的（p < 0.05）。对照组、TGF-β1处理组和二甲双胍处理组之间的活死细胞染色无差异。总之，我们的研究表明，TGF-β1诱导EPCs中纤维化生物标志物的表达，二甲双胍治疗可减弱这种表达。因此，二甲双胍可能具有改善心血管疾病中EPC功能的治疗潜力。
Endothelial progenitor cells (EPCs) are a group of circulating cells with important functions in vascular repair and treatment of cardiovascular diseases. However, in patients with atrial fibrillation (AF), the number and function of EPCs reportedly are decreased. TGF-β is highly expressed in AF patients. In this study, we examined the effect of TGF-β1 on EPCs and the therapeutic outcome of metformin treatment on TGF-β1-induced EPCs. EPCs were induced with TGF-β1 at different concentrations (5 ng/ml, 10 ng/ml, and 20 ng/ml) for 48 h followed by western blot, qPCR, and immunofluorescence analyses to investigate changes in the levels of the fibrosis-related proteins, α-SMA, Col I, Col III, CTGF, and MMP-1. Live-dead cell staining was used to evaluate cell apoptosis. Compared with the control, TGF-β1 treatment significantly (p < 0.05) enhanced the levels of α-SMA, Col I, Col III, CTGF, and MMP-1 in a dose-dependent manner. The most effective concentration of TGF-β1 (20 ng/ml) was then used to induce fibrosis biomarker expression in EPCs, followed by treatment with metformin at different concentrations (0.5, 1, and 2 mmol/l). Metformin treatment suppressed TGF-β-induced expression of all above factors, with the effect at 2 mmol/l being significant (p < 0.05). Live-dead cell staining showed no difference among the control, TGF-β1-treated, and metformin-treated groups. In conclusion, our study showed that TGF-β1 induces the expression of fibrosis biomarkers in EPCs, which is attenuated by treatment with metformin. Thus, metformin may have therapeutic potential for improving EPC function in cardiovascular diseases.