microRNA-133b在低切应力诱导血管内皮细胞影响血管平滑肌细胞增殖中的作用  

The role of microRNA-133b in proliferation of vascular smooth muscle cells induced by endothelial cells under low shear stress

作　　者：马英英[1] 王璐[1] 包晗[1] 韩悦[1] 齐颖新[1] 

机构地区：[1]上海交通大学生命科学技术学院,力学生物学研究所,上海200240

出　　处：《医用生物力学》2016年第5期408-415,共8页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(11002091,11222223,11232010)

摘　　要：目的研究血管内皮细胞(endothelial cells,ECs)直接感受低切应力刺激后分泌类胰岛素生长因子-1(insulinlike growth factor-1,IGF-1)影响血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖这一过程中microRNAs(miRs)的作用。方法用平行平板流动腔系统对ECs施加1.5 Pa正常切应力(normal shear stress,NSS)和0.5 Pa低切应力(low shear stress,Low SS),Real-time PCR检测VSMCs的miRs变化。用miRs预测网站预测miR-133b靶基因并验证。Western blotting检测核糖核酸结合蛋白1(polypyrimidine tract binding protein 1,Ptbp1)和N-myc下游调节基因1(N-myc downstream regulated 1,Ndrg1)的蛋白水平变化。Ed U流式检测miR-133b对VSMCs增殖的影响。结果 IGF-1静态刺激后,VSMCs的miR-133b和miR-378a表达上升。Low SS条件下,VSMCs的miR-133b表达显著上升,miR-378a表达无明显变化。下调VSMCs的miR-133b表达,Ptbp1、Ndrg1的mRNA水平均显著升高,上调VSMCs的miR-133b的表达,Ptbp1、Ndrg1的mRNA和蛋白水平显著降低,并且显著促进VSMCs增殖。结论在Low SS条件下ECs分泌IGF-1可能通过调控联合培养VSMCs的miR-133b和靶基因Ptbp1和Ndrg1促进VSMCs增殖。研究结果为心血管疾病治疗提供了一个新的潜在靶标。Objective To investigate the role of microRNAs （miRs） in the proliferation of vascular smooth muscle cells （VSMCs）induced by endothelial insulin-like growth factor-1 （IGF-1） under low shear stress （LowSS）. Methods Endothelial cells （ECs） and VSMCs were co-cultured and exposed to normal shear stress （NSS, 1. 5 Pa） and LowSS C0.-5 Pa） for 12 h with parallel plate flow chamber system, respectively. Real-time PCR was used to examine the expression levels of miRs. The target genes of miR-133b were predicted by multiple algo- rithms. The expression of polypyrimidine tract binding protein I （Ptbp]） and N-myc downstream regulated 1 （Ndrgl） in VSMCs was detected by Western blotting. The VSMC proliferation was detected by EdU flow cytome- try assay. Results After treated with recombinant IGF-I, the expression of both miR-133b and miR-378a in VSMCs was increased. Compared with NSS, LowSS significantly induced the expression of miR-133b in the co- cultured VSMCs, but had no obvious effect on miR-378a. In VSMCs, the protein and mRNA levels of Ptbpl and Ndrgl were down-regulated by miR-133b mimics, miR-133b inhibitor up-regulated the mRNA levels of Ptbpl andNdrgl. miR-133b overexpression promoted the proliferation of VSMCs significantly. Conclusions IGF-1 secreted by ECs in response to LowSS can upregulate the expression of miR-133b in the co-cultured VSMCs, which sub- sequently depresses the expression of Ptbpl and Ndrgl, and induces the proliferation of VSMCs eventually. The research findings provide a potential new target for cardiovascular disease therapy.

关 键 词：切应力 内皮细胞 血管平滑肌细胞 类胰岛素生长因子-1 细胞增殖 

分 类 号：R318.01[医药卫生—生物医学工程]