Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province , Hunan University Changsha , Hunan , 410082 , China.
Institute of Molecular Medicine (IMM), Renji Hospital, Shanghai Jiao Tong University School of Medicine, and College of Chemistry and Chemical Engineering , Shanghai Jiao Tong University , Shanghai 200240 , China.
Anal Chem. 2019 May 21;91(10):6675-6681. doi: 10.1021/acs.analchem.9b00609. Epub 2019 May 1.
Stroke is one of the leading causes of disability and death among adults worldwide and results in numerous biochemical alterations. However, few efficient biomarkers are clinically available to diagnose stroke because of the limitations of biomarkers and their probes. In this work, we utilized frozen brain slices of middle cerebral artery occlusion (MCAO) in a mouse model of ischemia to select a specific binding aptamer, termed LCW17, by tissue-based SELEX (systematic evolution of ligands by exponential enrichment). LCW17 was enhanced in binding in ischemic brain slices compared to sham control. We identified the binding target of LCW17 as vigilin. Vigilin is increased in ischemia brain slices and exhibits enhanced release from cultured hippocampal neurons after oxygen glucose deprivation in vitro. Taken together, ischemic brain slice-based aptamer selection will enable identification of more probes and potential target molecules for diagnosis and therapy of ischemic stroke. Aptamer LCW17 and vigilin may potentially be applied to define the molecular mechanism underlying ischemic stroke, as well as its diagnosis.
中风是全球成年人残疾和死亡的主要原因之一，会导致许多生化改变。然而，由于生物标志物及其探针的局限性，临床上可用的有效生物标志物很少。在这项工作中，我们利用缺血性中风小鼠模型中的冷冻大脑切片，通过基于组织的 SELEX（指数富集的配体系统进化）选择了一种特定的结合适体，称为 LCW17。与假手术对照相比，LCW17 在缺血性脑切片中的结合增强。我们确定了 LCW17 的结合靶标为vigilin。vigilin 在缺血性脑切片中增加，并在体外缺氧葡萄糖剥夺后从培养的海马神经元中增强释放。总之，基于缺血性脑切片的适体选择将能够鉴定更多的探针和潜在的靶分子，用于缺血性中风的诊断和治疗。适体 LCW17 和 vigilin 可能潜在地应用于定义缺血性中风的分子机制及其诊断。
