Chen Chao, Cheng Guangqing, Yang Xiaoni, Li Changsheng, Shi Ran, Zhao Ningning
Department of Traditional Chinese Medicine, Shandong Qianfoshan Hospital Jinan 250014, China.
Am J Transl Res. 2016 Jul 15;8(7):2981-91. eCollection 2016.
Endothelial cell (EC) apoptosis is a crucial process for the development of atherosclerosis. Tanshinol is reported to protect vascular endothelia and attenuate the formation of atherosclerosis. However, the potential molecule mechanism of the protective role of tanshinol in atherosclerosis need to be further investigated. ApoE(-/-)mice were fed with a high-fat diet and treated with tanshinol to detect the effect of tanshinol on endothelial cells apoptosis with TUNEL staining assay. qRT-PCR and Western blot were performed to examine the expression of TUG1 and miR-26a in endothelial cells. RNA-binding protein immunoprecipitation assay was performed to verify the relationship between TUG1 and miR-26a. It has been shown that tanshinol reduced the aortic atherosclerotic lesion area in the entire aorta and aortic sinus in a concentration dependent manner, and suppressed the endothelial cells apoptosis in ApoE(-/-) mice. We further found that the mRNA level of TUG1 was reduced and the expression of miR-26a was up-regulated by tanshinol in endothelial cells. In addition, TUG1 down-regulated the expression of miR-26a in ECV304 cells. Finally, it was shown that overexpression of TUG1 removed the reversed effect of tanshinol on oxidized low-density lipoprotein (ox-LDL)-induced endothelial cells apoptosis. Taken together, our study reveals that tanshinol could attenuate the endothelial cells apoptosis in atherosclerotic ApoE(-/-) mice. Moreover, low TUG1 expression and high level of miR-26a are associated with the endothelial protecting effect of tanshinol.
内皮细胞(EC)凋亡是动脉粥样硬化发展的关键过程。据报道,丹参素可保护血管内皮并减轻动脉粥样硬化的形成。然而,丹参素在动脉粥样硬化中发挥保护作用的潜在分子机制仍需进一步研究。给载脂蛋白E基因敲除(ApoE(-/-))小鼠喂食高脂饮食并用丹参素进行处理,通过TUNEL染色法检测丹参素对内皮细胞凋亡的影响。采用实时定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法(Western blot)检测内皮细胞中TUG1和微小RNA-26a(miR-26a)的表达。进行RNA结合蛋白免疫沉淀试验以验证TUG1与miR-26a之间的关系。结果表明,丹参素以浓度依赖性方式减少了整个主动脉和主动脉窦的主动脉粥样硬化病变面积,并抑制了ApoE(-/-)小鼠的内皮细胞凋亡。我们进一步发现,丹参素可降低内皮细胞中TUG1的mRNA水平并上调miR-26a的表达。此外,TUG1下调了ECV304细胞中miR-26a的表达。最后,结果表明,TUG1的过表达消除了丹参素对氧化型低密度脂蛋白(ox-LDL)诱导的内皮细胞凋亡的逆转作用。综上所述,我们的研究表明,丹参素可减轻动脉粥样硬化ApoE(-/-)小鼠的内皮细胞凋亡。此外,低水平的TUG1表达和高水平的miR-26a与丹参素的内皮保护作用相关。