微小RNA-92a通过激活SMAD2/3/FAK/Akt/eNOS信号通路靶向生长分化因子11来调控内皮祖细胞。
MiR-92a regulates endothelial progenitor cells (EPCs) by targeting GDF11 via activate SMAD2/3/FAK/Akt/eNOS pathway.
作者信息
Huang Hai-Tao, Liu Zhen-Chuan, Wu Kai-Qin, Gu Shao-Rui, Lu Tian-Cheng, Zhong Chong-Jun, Zhou Yong-Xin
机构信息
Department of Thoracic and Cardiovascular Surgery, Nantong First People's Hospital, Nantong 226001, China.
Department of Thoracic and Cardiovascular Surgery, Tongji Hospital, School of Medicine, Tongji University, Shanghai 200065, China.
出版信息
Ann Transl Med. 2019 Oct;7(20):563. doi: 10.21037/atm.2019.09.35.
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-luciferase 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 GDF11 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 miR-92a on EPCs are mediated by regulating of GDF11 and downstream SMAD2/3/FAK/Akt/eNOS signaling pathway.
背景
miR-92a对内皮祖细胞（EPCs）的影响仍未完全阐明。本研究旨在通过靶向生长分化因子11（GDF11），探讨miR-92a在缺氧（HO）或高糖（HG）诱导的EPCs损伤模型中对EPCs的影响。
方法
首先研究miR-92a对HO或HG处理的EPCs的影响。随后，阐明miR-92a通过靶向GDF11对EPCs的作用机制。分别用MTT法、流式细胞术、Transwell法、管腔形成实验检测细胞增殖、凋亡、迁移和血管生成能力。24小时后，通过荧光强度检测活性氧（ROS）水平。用ELISA法测定乳酸脱氢酶（LDH）和一氧化氮（NO）水平。通过免疫荧光检测血管内皮生长因子受体-1（FLK-1）和血管性血友病因子（vWF）的表达。分别用PCR和蛋白质印迹法检测mRNA和蛋白质表达水平。通过双荧光素酶报告基因检测法评估miR-92a与GDF11之间的相互作用。
结果
我们的结果表明，HO或HG可增加EPCs的凋亡、LDH产生和ROS生成，但降低其迁移能力、管腔形成能力和NO生成；抑制miR-92a可减轻HO或HG诱导的EPCs损伤，而miR-92a过表达则产生相反的效果；GDF11小干扰RNA可逆转抑制miR-92a对EPCs的保护作用，过表达GDF11可挽救miR-92a过表达对EPCs的有害作用，这表明miR-92a的有害作用与其抑制GDF11及随后使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。miR-92a对EPCs的影响是通过调节GDF11及下游SMAD2/3/FAK/Akt/eNOS信号通路介导的。
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