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

摘要 外周动脉疾病（PAD）影响了850万40岁以上的美国人。最近有证据表明 研究和其他研究表明，氧化应激在PAD的病理生理学中起着重要作用， 行走障碍和生活质量下降。很少有治疗方法可以改善行走 PAD患者的距离和生活质量。PAD的一种新兴治疗方法是使用 微RNA（miR）。miRs是一种内源性的21 - 25个核苷酸的非编码RNA，可以调节 转录后基因表达miR最常见的作用机制是通过结合3'端未取代的 在一些实施方案中，所述方法包括抑制靶mRNA的翻译区，从而降低mRNA表达或蛋白质翻译。循环 miRNAs代表了PAD诊断和预后的潜在生物标志物，也是PAD诊断和预后的起点。 个性化治疗。最近在PAD和后肢缺血模型中的证据已经将miR-210鉴定为 低氧条件下基因表达的主要调节因子。我们实验室的初步工作 证明血清和腓肠肌样品中的miR-210增加，并且与 疾病进展。此外，我们已经确定，血运重建手术可以减少循环， PAD患者术后6个月血清中miR-210水平。已经显示miR-210可以 负调节线粒体呼吸活性，并通过抑制 ISCU（铁硫簇支架同源物）和COX 10（细胞色素c氧化酶组装蛋白）。所以我们 一个中心假设是，miR-210基因表达是氧化应激的主要调节因子， 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的miR-210基因表达与健康人不同 年龄匹配的对照，并与氧化应激，氧化代谢，线粒体功能，步行 功能和生活质量。 目的#2：血管内和开放旁路血运重建术可通过以下方式调节氧化应激： 降低PAD患者腓肠肌和血清中miR-210的表达， 功能、氧化代谢、行走功能和生活质量。 目的#3：在PAD的新型常氧/缺氧/高氧模型中利用体外研究，以确定 通过诱导/抑制miR-210基因表达及其与mRNA的相互作用改变基因表达 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.