Wu W, Shang Y Q, Dai S L, Yi F, Wang X C
Bratisl Lek Listy. 2017;118(8):499-503. doi: 10.4149/BLL_2017_096.
Vascular calcification is one of the most important factors for high morbidity and mortality from cardiovascular and cerebrovascular diseases. The aim of this study is to investigate the effect and mechanism of miR-26a on vascular smooth muscle cell calcification. First, the VSMCs were induced by β-glycerol phosphate (β-GP) for 7d and 14d, and Alizarin Red S staining was performed to examine the mineralized nodule change; then real time RT-PCR and western blotting were performed to explore the expression of miR-26a, CTGF, OPG, RANKL and ALP in un-induced and β-GP-induced VSMCs; next, the VSMCs were transfected with miR-26a mimics, and Alizarin Red S staining was performed to examine the mineralized nodule change; finally, real time RT-PCR and western blotting were performed to explore the expression of miR-26a, CTGF, OPG, RANKL and ALP in un-transfected and miR-26a mimics transfected VSMCs. After β-GP treatment, β-GP promoted clear mineralized nodule changes, and miR-26a and OPG expression were significantly decreased and CTGF, RANKL and ALP expression were increased in VSMCs. Overexpression of miR-26a inhibited VSMCs calcification induced by β-GP, and regulated the expression of CTGF, OPG, RANKL and ALP. Our findings suggested that up-regulation of miR-26a before β-GP treatment inhibits VSMCs calcification through targeting CTGF (Fig. 4, Ref. 18).
血管钙化是导致心血管和脑血管疾病高发病率和高死亡率的最重要因素之一。本研究旨在探讨miR-26a对血管平滑肌细胞钙化的影响及其机制。首先，用β-甘油磷酸（β-GP）诱导血管平滑肌细胞7天和14天，然后进行茜素红S染色以检测矿化结节的变化；接着进行实时RT-PCR和蛋白质印迹法，以探究未诱导和β-GP诱导的血管平滑肌细胞中miR-26a、结缔组织生长因子（CTGF）、骨保护素（OPG）、核因子κB受体活化因子配体（RANKL）和碱性磷酸酶（ALP）的表达；接下来用miR-26a模拟物转染血管平滑肌细胞，再进行茜素红S染色检测矿化结节的变化；最后，进行实时RT-PCR和蛋白质印迹法，以探究未转染和miR-26a模拟物转染的血管平滑肌细胞中miR-26a、CTGF、OPG、RANKL和ALP的表达。β-GP处理后，β-GP促进了明显的矿化结节变化，血管平滑肌细胞中miR-26a和OPG的表达显著降低，而CTGF、RANKL和ALP的表达增加。miR-26a的过表达抑制了β-GP诱导的血管平滑肌细胞钙化，并调节了CTGF、OPG、RANKL和ALP的表达。我们的研究结果表明，在β-GP处理前上调miR-26a可通过靶向CTGF抑制血管平滑肌细胞钙化（图4，参考文献18）。
