Vascular calcification (VC) is a vital factor for cardiovascular morbidity and mortality. Accumulating data suggest that microRNA (miR) is implicated in the VC. The main purpose of this study is to study the influence of miR-155-5p overexpression on VC development in vitro and in vivo. Immunofluorescence staining, real-time PCR, alizarin red staining, alkaline phosphatase (ALP) activity assay, western blot, luciferase assay, hematoxylin-eosin (HE), Masson's trichrome staining, and calcium content assay were used in this research. The results showed that miR-155-5p was decreased in the rat vascular smooth muscle cells (rVSMCs) undergoing calcification in vitro. MiR-155-5p overexpression reversed the increase of calcification and ALP activity in calcified cells. Further, overexpression of miR-155-5p inhibited the transforming growth factor-β1 (TGF-β1)/Smad2/3 signaling pathway, as evidenced by decreased protein expression of TGF-β1, pSmad-2 and pSmad-3 in rVSMCs. MiR-155-5p was showed to target Smad2 directly. Moreover, miR-155-5p upregulation reduced vascular thickening, fibrosis and calcium content of aorta abdominalis in CaCl2-mediated VC model. Collectively, our results suggest that miR-155-5p overexpression may inhibit VC development through suppressing TGF-β1/Smad2/3 signaling pathway in vivo and in vitro, indicating that miR-155-5p may act as a potential therapeutic target for VC-related disease.

中文翻译：

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microRNA-155-5p对血管钙化的抑制作用伴随着TGF-β1/ Smad2 / 3信号通路的失活。

血管钙化（VC）是心血管疾病和死亡率的重要因素。越来越多的数据表明，microRNA（miR）与VC有关。这项研究的主要目的是研究miR-155-5p过表达对体外和体内VC发育的影响。本研究使用了免疫荧光染色，实时荧光定量PCR，茜素红染色，碱性磷酸酶（ALP）活性测定，蛋白质印迹，荧光素酶测定，苏木精-伊红（HE），马森三色染色和钙含量测定。结果表明，miR-155-5p在体外钙化的大鼠血管平滑肌细胞（rVSMC）中降低。MiR-155-5p过表达逆转了钙化细胞中钙化和ALP活性的增加。进一步，miR-155-5p的过表达抑制了转化生长因子-β1（TGF-β1）/ Smad2 / 3信号传导途径，rVSMCs中TGF-β1，pSmad-2和pSmad-3的蛋白表达降低证明了这一点。已显示MiR-155-5p直接靶向Smad2。而且，在CaCl2介导的VC模型中，miR-155-5p的上调减少了血管增厚，纤维化和腹主动脉的钙含量。总体而言，我们的结果表明，miR-155-5p的过表达可能通过抑制体内和体外的TGF-β1/ Smad2 / 3信号通路来抑制VC的发展，表明miR-155-5p可能是VC-的潜在治疗靶标相关疾病。在CaCl2介导的VC模型中，miR-155-5p的上调减少了血管增厚，纤维化和腹主动脉的钙含量。总体而言，我们的结果表明，miR-155-5p的过表达可能通过抑制体内和体外的TGF-β1/ Smad2 / 3信号通路来抑制VC的发展，表明miR-155-5p可能是VC-的潜在治疗靶标相关疾病。在CaCl2介导的VC模型中，miR-155-5p的上调减少了血管增厚，纤维化和腹主动脉的钙含量。总体而言，我们的结果表明，miR-155-5p的过表达可能通过抑制体内和体外的TGF-β1/ Smad2 / 3信号通路来抑制VC的发展，表明miR-155-5p可能是VC-的潜在治疗靶标相关疾病。