The role of microRNA-210 in regulating oxidative stress in patients with peripheral artery disease

摘要 40岁以上的美国人中有850万人患有外周动脉疾病。最近来自OUT的证据 工作和其他研究表明，氧化应激在PAD的病理生理学中的中心作用及其相关性 有更大的行走障碍和生活质量下降。很少有治疗方法能改善行走 PAD患者的距离和生活质量。一种新的治疗PAD的方法是使用 MircroRNAs(MiR)。MIR是一种内源性的21∼非编码核糖核酸，可以调节 转录后基因表达。MiRs最常见的作用机制是通过与3‘Un结合而发挥作用。 目的基因的翻译区域，从而减少了mRNA的表达或蛋白质的翻译。流通中 MiRNAs是PAD诊断和预后的潜在生物标志物，也是PAD的一个起点 个体化治疗。最近在PAD和后肢缺血模型中的证据表明miR-210是一种 掌握低氧条件下基因表达的调节。我们实验室的初步工作已经完成 结果表明，血清和腓肠肌标本中miR-210水平升高，且与 疾病的发展。此外，我们发现血运重建手术可以减少血液循环。 PAD患者术后6个月血清MIR-210水平。已有研究表明，miR-210可以 抑制线粒体呼吸活性，增加活性物质生成 ISCU(铁硫簇支架同系物)和COX10(细胞色素C氧化酶组装蛋白)。因此，我们的 中心假设是miR-210基因表达是氧化应激的主要调节因子，并与 线粒体功能障碍、氧化代谢、行走功能和生活质量。 目的#1：PAD患者腓肠肌和血清中MIR-210基因的表达与健康人不同 年龄与对照组相匹配，并与氧化应激、氧化代谢、线粒体功能、步行相关 功能和生活质量。 目的#2：血管内和开放搭桥血管重建术可以通过以下方式调节氧化应激 降低PAD患者腓肠肌和血清miR-210的表达及改善线粒体 功能、氧化代谢、行走功能和生活质量。 目的#3：利用一种新的PAD常氧/低氧/高氧模型的体外研究来确定PAD的程度 通过诱导/抑制miR-210基因表达及其与mRNA的相互作用改变哪些基因的表达 表情。

Abstract Peripheral artery disease (PAD) affects 8.5 million of Americans over 40 years of age. Recent evidence from out work and others suggest the central role of oxidative stress in the pathophysiology of PAD and its association with greater walking impairment and decline in quality of life. Few therapeutic treatments can improve walking distances and quality of life in PAD patients. A new emerging therapeutic approach for PAD is the usage of mircroRNAs (miRs). miRs are endogenous 21∼25 nucleotides noncoding RNA, that can regulate posttranscriptional gene expression. The most common mechanism of action of miRs is by binding to the 3' un- translated region of a target mRNA and thereby reducing mRNA expression or protein translation. Circulating miRNAs, represent potential biomarkers for the diagnosis and prognosis of PAD and a starting point for individualized treatment. Recent evidence in PAD and hindlimb ischemia models have identified miR-210 as a master regulator of gene expression under hypoxic conditions. Preliminary work from our laboratory has demonstrated that miR-210 in the serum and gastrocnemius samples is increased and positively correlated with disease progression. Furthermore, we have identified that revascularization operations can decrease circulating miR-210 in the serum of PAD patients six-months after the operation. It has been shown that miR-210 can negatively regulate mitochondrial respiratory activity and increase reactive species generation by inhibiting the ISCU (iron-sulfur cluster scaffold homolog) and COX10 (cytochrome c oxidase assembly protein). Thus, our central hypothesis, is that miR-210 gene expression is a master regulator of oxidative stress and is associated with mitochondrial dysfunction, oxidative metabolism, walking function and quality of life. Aim #1: miR-210 gene expression in the gastrocnemius and serum of patients with PAD, is different than healthy age matched controls, and correlates with oxidative stress, oxidative metabolism, mitochondrial function, walking function and quality of life. Aim #2: Endovascular and open bypass revascularization procedures can regulate oxidative stress by decreasing miR-210 expression in the gastrocnemius and serum of PAD patients and improve mitochondrial function, oxidative metabolism, walking function and quality of life. Aim #3: Utilize in-vitro studies in a novel normoxia/hypoxia/hyperoxia model of PAD to determine the extent to which gene expression changes by inducing/inhibiting miR-210 gene expression and its interactions with mRNA expression.