Department of Cardiology, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150001, People's Republic of China.
Department of Clinical Laboratory, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150001, People's Republic of China.
Acta Histochem. 2020 May;122(4):151551. doi: 10.1016/j.acthis.2020.151551. Epub 2020 Apr 25.
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 CaCl-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.
血管钙化（VC）是心血管发病率和死亡率的重要因素。越来越多的证据表明 microRNA（miR）参与了 VC 的发生。本研究的主要目的是研究 miR-155-5p 过表达对体外和体内 VC 发展的影响。本研究采用免疫荧光染色、实时 PCR、茜素红染色、碱性磷酸酶（ALP）活性测定、western blot、荧光素酶测定、苏木精-伊红（HE）染色、Masson 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（rVSMCs）中表达下调。miR-155-5p 过表达逆转了钙化细胞中钙化和 ALP 活性的增加。此外，miR-155-5p 过表达抑制了转化生长因子-β1（TGF-β1）/Smad2/3 信号通路，rVSMCs 中 TGF-β1、pSmad-2 和 pSmad-3 的蛋白表达降低。结果表明 miR-155-5p 可直接靶向 Smad2。此外，miR-155-5p 的上调减少了 CaCl 介导的 VC 模型中腹主动脉的血管增厚、纤维化和钙含量。综上所述，我们的结果表明，miR-155-5p 过表达可能通过抑制 TGF-β1/Smad2/3 信号通路在体内和体外抑制 VC 的发展，表明 miR-155-5p 可能是 VC 相关疾病的潜在治疗靶点。
