MiR-92a regulates endothelial progenitor cells (EPCs) by targeting GDF11 via activate SMAD2/3/FAK/Akt/eNOS pathway
MiR-92a regulates endothelial progenitor cells (EPCs) by targeting GDF11 via activate SMAD2/3/FAK/Akt/eNOS pathway
复制标题
MiR-92a 通过激活 SMAD2/3/FAK/Akt/eNOS 通路靶向 GDF11 来调节内皮祖细胞 (EPC)
DOI：
10.21037/atm.2019.09.35
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发表时间：
2019-10-01
影响因子：
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通讯作者：
Zhou, Yong-Xin
中科院分区：
文献类型：
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作者：
Huang, Hai-Tao;Liu, Zhen-Chuan;Zhou, Yong-Xin
背景：miR-92a对EPCs的影响尚不清楚。本研究旨在通过靶向GDF11（分化生长因子11），探讨miR-92a在缺氧（HO）或高糖（HG）诱导的EPCs损伤模型中对EPCs（内皮祖细胞）的影响。方法：首先研究miR-92a对HO或HG后EPCs的影响。随后，我们阐明了miR-92a通过靶向GDF11对EPCs的作用机制。分别用MTT、流式细胞术、transwell、成管法检测细胞增殖、细胞凋亡、细胞迁移、血管生成。24 h后，用荧光强度测定活性氧（ROS）水平。ELISA法检测LDH、NO（一氧化氮）水平。免疫荧光法检测胎儿肝激酶1 （FLK-1）和血管性血友病因子（vWF）的表达。分别用PCR和western blotting检测mRNA和蛋白的表达水平。通过双荧光素酶报告基因法评估miR-92a与GDF11之间的相互作用。结果：我们的研究结果表明，HO或HG增加了EPCs的凋亡、LDH的产生和ROS的产生，但降低了EPCs的迁移和成管能力以及NO的产生；抑制miR-92a可降低HO或hg诱导的EPCs损伤，而过表达miR-92a则具有相反的作用；抑制miR-92a对EPCs的保护作用可以被GDFH siRNA逆转，过表达miR-92a诱导的有害作用可以通过过表达GDF11来挽救，这表明miR-92a的有害作用与其抑制GDF11并随后使SMAD2/3/FAK/Akt/eNOS信号通路失活有关。结论：抑制miR-92a可保护EPCs免受HO或hg诱导的损伤。miR92a对EPCs的作用是通过调控GDFH和下游SMAD2/3/FAK/Akt/eNOS信号通路介导的。
Background: The effects of miR-92a on EPCs are still poorly elucidated. This study aimed to investigate the effects of miR-92a on EPCs (Endothelial progenitor cells) in a model of hypoxia (HO) or high glucose (HG)-induced EPCs injury by targeting GDF11 (Differentiation growth factor 11).Methods: The effects of miR-92a on EPCs subjected to HO or HG were investigated firstly. Subsequently, the action mechanism of miR-92a on EPCs by targeting GDF11 was elucidated. Proliferation, apoptosis, migration, angiogenesis was measured with MTT, flow cytometry, transwell, tube formation respectively. After 24 h, levels of reactive oxygen species (ROS) were measured by fluorescence intensity. LDH and NO (nitric oxide) levels were determined by ELISA. The expression of FLK-1 (fetal liver kinase 1) and vWF (von Willebrand factor) was detected by immunofluorescence. mRNA and protein expression levels were examined using PCR and western blotting respectively. The interaction between miR-92a and GDF11 was evaluated by dual-lucifcrase reporter assay.Results: Our results showed that HO or HG increased apoptosis, production of LDH and generation of ROS, but decreased the ability of migration and tube formation and generation of NO in EPCs; inhibiting of miR-92a decreased HO or HG-induced injury of EPCs, whereas miR-92a over-expression had the opposite effect; the protective effects induced by inhibiting of miR-92a on EPCs could be reversed by GDFH siRNA and the harmful effects induced by over-expression of miR-92a could be rescued by over-expression of GDF11, which showed that the harmful effects of miR-92a be related to its inhibition of GDF11 and subsequent inactivation of the SMAD2/3/FAK/Akt/eNOS signaling pathway.Conclusions: Inhibiting miR-92a can protect EPCs from HO or HG-induced injury. The effect of miR92a on EPCs are mediated by regulating of GDFH and downstream SMAD2/3/FAK/Akt/eNOS signaling pathway.