北京大学学报（医学版）2016，Vol.48Issue(5)：756-765,10.DOI:10.3969/j.issn.1671-167X.2016.05.002
microRNA 在高磷诱导血管平滑肌细胞钙化早期的动态变化
Dynamic alteration of microRNA in high phosphorus induced calcification of vascular smooth muscle cell
摘要
Abstract
Objective:To study the change of microRNA during the early stage of high phosphorus in-duced vascular smooth muscle cell (VSMC)calcification and its related mechanism.Methods:The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol /L)for 7 d.The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content,real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules.Based on the cell calcification model,micro-RNA microarray array was applied to screen the profiles of microRNA expression in VSMC following high Pi stimulation for different periods (0,3 and 12 h).The array data were analyzed by TAMtool to explore the activated signaling pathway.Results:The calcium content of A7r5 cells induced by high Pi was in-creased 9.6 times high as cells without Pi treatment (P <0.05 ).VSMC contractile phenotype genes (SM-αactin,SM22)were down-regulated (P <0.05 ),while calcification-related genes (BMP2, MSX2,Runx2)were up-regulated (P <0.05)in VSMC stimulated by high Pi.The calcium nodules were obviously formed in cells after 7 d high Pi treatment.In microarray experiment,680 individual mi-croRNAs were detected in high Pi-treated VSMCs at different time points (0,3 and 12 h).Among these genes,miR-183,miR-664 and miR-9 * were increased whereas miR-542-5P,let-7f and miR-29a were decreased in time-dependent manners.Twenty-six kinds of signaling pathways,including cell apoptosis, differentiation and proliferation,were significantly activated.All these activated pathways were associated with calcification.Conclusion:This study implies that microRNA changed in high Pi-induced VSMCs may involve in the process of calcification.关键词
血管钙化/肌,平滑,血管/微RNA/信号转导Key words
Vascular calcification/Muscle,smooth,vascular/microRNA/Signal transduction分类
医药卫生引用本文复制引用
肖洋,杜瑶瑶,高成,孔炜..microRNA 在高磷诱导血管平滑肌细胞钙化早期的动态变化[J].北京大学学报（医学版）,2016,48(5):756-765,10.基金项目
国家基金委重大国际合作项目（81220108004）、国家重点基础研究发展计划（973计划，2012CB518002）、国家自然科学基金（81070243、81121061、91339000）和国家杰出青年基金（81225002）资助 Supported by the Foundation of Major International Cooperation （81220108004），the National Basic Research Program of China （973 Program，2012CB518002），the National Natural Science Foundation of China （81070243，81121061，91339000）and the National Science Fund for Distinguished Young Scholars （）
