Downregulation of miR-542-3p promotes osteogenic transition of vascular smooth muscle cells in the aging rat by targeting BMP7
Downregulation of miR-542-3p promotes osteogenic transition of vascular smooth muscle cells in the aging rat by targeting BMP7
复制标题
下调miR-542-3p通过靶向BMP7促进衰老大鼠血管平滑肌细胞的成骨转变
DOI：
10.1186/s40246-019-0245-z
复制
发表时间：
2019-12-11
期刊：
影响因子：
4.5
通讯作者：
Qian, Dehui
中科院分区：
文献类型：
--
作者：
Liu, Huan;Wang, Hongwei;Qian, Dehui
背景：衰老被认为与心血管疾病密切相关，导致血管发生各种病理改变，包括血管细胞表型转变。在衰老的血管中，微小RNA（miRNA）介导的调节血管平滑肌细胞（VSMC）表型的机制仍不明确。采用miRNA微阵列比较老年大鼠（oVSMCs）和年轻大鼠（yVSMCs）的血管平滑肌细胞中miRNAs的表达。采用定量逆转录实时聚合酶链反应（qRT - PCR）和小RNA转染在体外探究oVSMCs和yVSMCs中miR - 542 - 3p的表达。通过β - 甘油磷酸酯（β - GP）处理诱导yVSMCs钙化。采用茜素红染色检测钙沉积。采用蛋白质印迹法和qRT - PCR研究平滑肌标志物平滑肌22α（SM22α）和钙调蛋白以及成骨标志物骨桥蛋白（OPN）和 Runt相关转录因子2（Runx2）的表达。采用慢病毒在yVSMCs中过表达miR - 542 - 3p和骨形态发生蛋白7（BMP7）。进行荧光素酶报告基因实验以确定miR - 542 - 3p的靶标。
结果：与yVSMCs相比，在oVSMCs中鉴定出28种下调和34种上调的miRNAs。qRT - PCR证实oVSMC表达的miR - 542 - 3p比yVSMCs低4倍。在yVSMCs中过表达miR - 542 - 3p抑制了β - GP诱导的成骨分化。此外，miR - 542 - 3p靶向BMP7，并且在表达miR - 542 - 3p的yVSMCs中过表达BMP7可逆转miR - 542 - 3p对成骨分化的抑制作用。
结论：miR - 542 - 3p通过靶向BMP7调节VSMCs的成骨分化，这表明oVSMCs中miR - 542 - 3p的下调在老年大鼠的成骨转变中起关键作用。
Background: Aging is believed to have a close association with cardiovascular diseases, resulting in various pathological alterations in blood vessels, including vascular cell phenotypic shifts. In aging vessels, the microRNA(miRNA)-mediated mechanism regulating the vascular smooth muscle cell (VSMC) phenotype remains unclarified. MiRNA microarray was used to compare the expressions of miRNAs in VSMCs from old rats (oVSMCs) and young rats (yVSMCs). Quantitative reverse transcription real-time PCR (qRT-PCR) and small RNA transfection were used to explore the miR-542-3p expression in oVSMCs and yVSMCs in vitro. Calcification induction of yVSMCs was conducted by the treatment of beta-glycerophosphate (beta-GP). Alizarin red staining was used to detect calcium deposition. Western blot and qRT-PCR were used to investigate the expression of the smooth muscle markers, smooth muscle 22a (SM22 alpha) and calponin, and the osteogenic markers, osteopontin (OPN), and runt-related transcription factor 2 (Runx2). Lentivirus was used to overexpress miR-542-3p and bone morphogenetic protein 7 (BMP7) in yVMSCs. Luciferase reporter assay was conducted to identify the target of miR-542-3p.Results: Compared with yVSMCs, 28 downregulated and 34 upregulated miRNAs were identified in oVSMCs. It was confirmed by qRT-PCR that oVSMC expressed four times lower miR-542-3p than yVSMCs. Overexpressing miR-542-3p in yVSMCs suppressed the osteogenic differentiation induced by beta-GP. Moreover, miR-542-3p targets BMP7 and overexpressing BMP7 in miR- 542-3p-expressing yVSMCs reverses miR-542-3p's inhibition of osteogenic differentiation.Conclusions: miR-542-3p regulates osteogenic differentiation of VSMCs through targeting BMP7, suggesting that the downregulation of miR-542-3p in oVSMCs plays a crucial role in osteogenic transition in the aging rat.