Monitoring Red Cell Metabolism using a Lab on a Chip

使用芯片实验室监测红细胞代谢

• 批准号: 7988641
• 负责人: Dana M Spence
• 金额: $ 7.49万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-03 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988641
• 关键词: Adenosine Triphosphate; Antioxidants; Biological Assay; Blood Circulation; Blood Pressure; Cell Line; Chemicals; Complications of Diabetes Mellitus; Detection; Development; Devices; Diabetes Mellitus; Endothelial Cells; Endothelium; Erythrocyte Deformability; Erythrocytes; Health; Hypertension; Literature; Measurement; Measures; Metabolic Pathway; Metabolism; Methods; Microcirculation; Microfluidic Microchips; Monitor; NADP; Nitric Oxide; Non-Insulin-Dependent Diabetes Mellitus; Oryctolagus cuniculus; Outcome; Oxygen; Patients; Pentosephosphate Pathway; Physiological; Production; Relaxation; Reporting; Resistance; Role; Scheme; Smooth Muscle Myocytes; System; Technology; Time; Tissues; base; diabetic; improved; micro-total analysis system

DESCRIPTION (provided by applicant): In this proposal, we describe the development of an analytical system that will enable the elucidation of the role of red blood cells (RBCs) in complications resulting from type II diabetes. Specifically, using a lab on a chip approach, we propose to examine the ability of erythrocytes to maintain inherent deformability via their antioxidant defense system. Recently, it has been demonstrated that erythrocyte deformability is a determinant of deformation-induced release of adenosine triphosphate (ATP). Moreover, this ATP is a known stimulant of nitric oxide production in endothelial cells that line resistance vessels in the microcirculation. When released, this endothelium-derived NO results in the eventual relaxation of smooth muscle cells surrounding the resistance vessels and subsequent dilation of the circulatory vessel. This dilation allows for an increase in erythrocyte flow delivering oxygen to required tissues and maintaining proper blood pressure. It has been reported in the literature that the erythrocytes of patients with type II diabetes have erythrocytes that are less deformable than erythrocytes of healthy patients. Thus, it is possible that the decreased deformability of erythrocytes from type II diabetics may be related to a decrease in erythrocyte-derived ATP (which in turn may result in diminished NO production in endothelial cells) and a subsequent increase in hypertension or circulation problems (both of which are complications suffered by most type II diabetics). Here, we propose 1)To develop an amperometric detection scheme on a microfluidic device to determine the ratio of GSH:GSSG in the RBCs of rabbits, 2) To develop an amperometric assay on a microfluidic device to determine the levels of NADPH in the RBCs of rabbits, 3) To develop a chip-based method for the determination of RBC deformability, 4) To employ the measurement schemes developed in 1-3 to determine the levels of those molecules while at the same time quantitatively measuring the amount of ATP released from rabbit RBCs and the RBCs from patients with Type II diabetes mellitus. Importantly, this device will not be employed solely for monitoring endpoint metabolites; rather, it will enable, for the first time, the ability to monitor a crucial metabolic pathway (specifically, the pentose phosphate pathway) in real time while simultaneously measuring the physical outcome (ATP release). Such determinations will help identify the role of RBCs in diabetic complications, thus improving the health of patients with diabetes.

描述（由申请人提供）：在本提案中，我们描述了分析系统的开发，该系统将能够阐明红细胞（RBC）在 II 型糖尿病引起的并发症中的作用。具体来说，我们建议使用芯片实验室方法来检查红细胞通过其抗氧化防御系统维持固有变形性的能力。最近，已经证明红细胞变形能力是变形诱导的三磷酸腺苷（ATP）释放的决定因素。此外，这种 ATP 是一种已知的刺激物，可促进微循环阻力血管内皮细胞中一氧化氮的产生。当释放时，这种内皮衍生的一氧化氮最终导致阻力血管周围的平滑肌细胞松弛，并随后扩张循环血管。这种扩张可以增加红细胞流量，将氧气输送到所需的组织并维持适当的血压。文献报道，II型糖尿病患者的红细胞比健康患者的红细胞变形程度更小。因此，II 型糖尿病患者红细胞变形能力的降低可能与红细胞来源的 ATP 减少（这反过来可能导致内皮细胞中 NO 产生减少）以及随后高血压或循环问题的增加（这两者都是大多数 II 型糖尿病患者所遭受的并发症）有关。在这里，我们建议 1) 在微流体装置上开发一种电流检测方案，以确定兔子红细胞中 GSH:GSSG 的比率，2) 在微流体装置上开发一种电流分析法，以确定兔子红细胞中 NADPH 的水平，3) 开发一种基于芯片的方法来测定红细胞变形性，4) 采用测量方法 1-3 中开发的方案确定这些分子的水平，同时定量测量兔红细胞和 II 型糖尿病患者红细胞释放的 ATP 量。重要的是，该设备不仅仅用于监测终点代谢物；相反，它将首次能够实时监测关键的代谢途径（特别是磷酸戊糖途径），同时测量身体结果（ATP 释放）。这些测定将有助于确定红细胞在糖尿病并发症中的作用，从而改善糖尿病患者的健康。

项目成果

Hydroxyurea stimulates the release of ATP from rabbit erythrocytes through an increase in calcium and nitric oxide production.

• DOI: 10.1016/j.ejphar.2010.07.012
• 发表时间: 2010-10-25
• 期刊: European journal of pharmacology
• 影响因子: 5
• 作者: Raththagala M;Karunarathne W;Kryziniak M;McCracken J;Spence DM
• 通讯作者: Spence DM

A Molecular Level Understanding of Zinc Activation of C-peptide and its Effects on Cellular Communication in the Bloodstream.

从分子水平了解 C 肽的锌激活及其对血流中细胞通讯的影响。

• DOI: 10.1900/rds.2009.6.148
• 发表时间: 2009
• 期刊: The review of diabetic studies : RDS
• 作者: Medawala,Wathsala;McCahill,Patrick;Giebink,Adam;Meyer,Jennifer;Ku,Chia-Jui;Spence,DanaM
• 通讯作者: Spence,DanaM

Development of an on-chip injector for microchip-based flow analyses using laminar flow.

开发用于使用层流进行基于微芯片的流动分析的片上注射器。

• DOI: 10.1039/b707410g
• 发表时间: 2007
• 期刊: Lab on a chip
• 影响因子: 6.1
• 作者: Moehlenbrock,MichaelJ;Martin,RScott
• 通讯作者: Martin,RScott

A microfluidic technique for monitoring bloodstream analytes indicative of C-peptide resistance in type 2 diabetes.

一种微流体技术，用于监测指示 2 型糖尿病 C 肽耐药性的血流分析物。

• DOI: 10.1039/b816740k
• 发表时间: 2009
• 期刊: The Analyst
• 作者: Oblak,TeresaD'Amico;Meyer,JenniferA;Spence,DanaM
• 通讯作者: Spence,DanaM

Merging Microfluidics with Micro-titre Technology for More Efficient Drug Discovery.

将微流控技术与微滴定技术相结合，实现更高效的药物发现。

• DOI: 10.1016/j.jala.2008.05.002
• 发表时间: 2008
• 期刊: JALA (Charlottesville, Va.)
• 作者: Tolan,NicoleV;Genes,LuizaI;Spence,DanaM
• 通讯作者: Spence,DanaM