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

摘要 外周动脉疾病 (PAD) 影响着 850 万 40 岁以上的美国人。最近的证据 工作和其他人表明氧化应激在 PAD 病理生理学及其关联中的核心作用 行走障碍更大，生活质量下降。很少有治疗方法可以改善步行 PAD 患者的距离和生活质量。一种新兴的 PAD 治疗方法是使用 微小RNA（miR）。 miRs是内源性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到 3' un- 目标 mRNA 的翻译区域，从而减少 mRNA 表达或蛋白质翻译。循环 miRNA 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 个体化治疗。 PAD 和后肢缺血模型的最新证据已确定 miR-210 是一种 缺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已 证明血清和腓肠肌样本中的 miR-210 增加并与 疾病进展。此外，我们已经发现血运重建手术可以减少循环 PAD患者术后六个月血清中的miR-210。研究表明，miR-210可以 通过抑制线粒体呼吸活性来负调节线粒体呼吸活性并增加活性物质的产生 ISCU（铁硫簇支架同源物）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，并且与 与线粒体功能障碍、氧化代谢、行走功能和生活质量有关。 目标#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、步行相关 功能和生活质量。 目标#2：血管内和开放式旁路血运重建手术可以通过以下方式调节氧化应激： 降低 PAD 患者腓肠肌和血清中 miR-210 的表达并改善线粒体 功能、氧化代谢、行走功能和生活质量。 目标#3：利用新型 PAD 正常氧/缺氧/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 的相互作用来改变基因表达 表达。

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.