miR-23a抑制肺微血管内皮细胞凋亡及在脓毒症急性肺损伤中的作用

项目名称： miR-23a抑制肺微血管内皮细胞凋亡及在脓毒症急性肺损伤中的作用

项目编号： No.81201448

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 阮溦

作者单位： 中南大学

项目金额： 23万元

中文摘要： 脓毒症急性肺损伤(ALI)是危重病患者死亡的主要原因。肺微血管内皮细胞（PMVEC）凋亡引起呼吸膜通透性增高，导致肺水肿、肺不张是ALI的病理基础。课题组前期工作证实了miR-23a通过在转录后水平抑制靶基因caspase-7和STK4的表达，减少脐静脉内皮细胞凋亡，提示miR-23a可能是脓毒症ALI中PMVEC凋亡调控的新靶点。本研究拟在体外培养的PMVEC中特异地沉默和过表达miR-23a，明确miR-23a对脂多糖诱导的小鼠PMVEC凋亡及对内皮细胞单层通透性的调控作用；并在miR-23a敲除和过表达的转基因小鼠体内进一步证实其对脓毒症时PMVEC凋亡及ALI的影响；最后在细胞模型，通过观察miR-23a对靶基因的作用，阐明其作用机制。本课题的实施不仅有助于从转录后水平揭示PMVEC凋亡的基因调控机制，也将为miR-23a作为脓毒症肺保护的新靶点提供坚实的理论基础和潜在的应用前景。

中文关键词： miR-23a；脓毒症；急性肺损伤；凋亡；肺微血管内皮细胞

英文摘要： Sepsis-indued acute lung injury (ALI) and acute distress syndrome (ARDS) are inflammatory disorders of the lung and are associated with high rates of morbidity and mortality. Apoptosis of pulmonary microvascular endothelial cells (PMVECs) contributes to the impairment of the alveolar-capillary membrane and leads to alveolar flooding with serum proteins and edema fluid. Our previous work found that miR-23a may attenuate TNF-α-induced endothelial cell apoptosis through down-regulation of the caspase-7 and serine/threonine kinase 4-caspase-3 pathways. Thus, we hypothesize that miR-23a may be involved in the development of ALI. We will first up-regulate (mimics) or down-regulate (inhibitor) endothelial miR-23a expression and evaluate the role of miR-23a in the lipopolysaccharides (LPS）-induced apoptosis of PMVECs and EC permeability in vitro. To inhibit miR-23a in vivo, we will design single-stranded RNA oligonucleotides complementary to miR-23a (antagomirs). To up-regulate miR-23a in vivo, we will design cholesterylated miR-23a mimics (agomirs). After successfully delivery of miR-23a antagomirs/miR-23a agomirs into mice, the regulation role of miR-23a in the development of ALI will be studied in vivo. We next sought to determine the effect of miR-23a on expression of the potential target genes. Our findings will su

英文关键词： miR-23a；sepsis；acute lung injury；apoptosis；pulmonary microvascular endothelial cell