Single-cell analysis of endothelial mechanotransduction mediated by endothelial surface glycocalyx

项目摘要/摘要 血管内皮细胞表面糖基化蛋白(ESG)是一种位于血管内皮细胞上的富含碳水化合物的细胞层。ESG是 由膜糖蛋白、糖胺多聚糖和蛋白多聚糖组成，形成块状的基质样物质 在血流的机械转导、内皮的维持等方面起关键作用的结构 通透性，控制白细胞黏附和炎症。最重要的常态之一 ESG的生理功能是介导导致钙离子摄取的机械转导和 血液流动引起的一氧化氮(NO)的产生。功能失调的ESG机械转导一直是 见于心血管疾病，如败血症、缺血再灌注、高血压和糖尿病。而当 ESG在心血管疾病中的重要作用已被证实，其生物力学特性，如 以及其正常机械转导的机制，一直拒绝阐明。这种缺乏 进展在很大程度上归功于这样一个事实，即分子和分子中发生的机械力和反应 亚细胞水平是短暂的、微小的，因此很难追踪和测量。当前的目标是 建议开发新的研究工具和模型系统，并用它们来揭示生物力学和 静电陀螺仪的机械传导特性。我们将描述ESG介导的潜在机制 一种新型原子力显微镜(AFM)--荧光在单细胞水平上的机械传递 显微镜方法。拟议的研究将实现对ESG中介的更清晰的理解 机械转导功能，对脓毒症等ESG相关疾病具有重要意义。 缺血再灌流、糖尿病和高血压及其治疗。

PROJECT SUMMARY/ABSTRACT Endothelial surface glycocalyx (ESG) is a carbohydrate-rich layer found on vascular endothelium. ESG is composed of membrane glycoproteins, glycosaminoglycans and proteoglycans, forming a bulky, matrix-like structure that serves critical functions in mechanotransduction of blood flow, maintenance of the endothelial permeability, and the control of leukocyte adhesion and inflammation. One of the most important normal physiological functions of ESG is to mediate mechanotransduction that leads to the intake of calcium ions and the production of Nitric Oxide (NO) in response to blood flow. Dysfunctional ESG mechanotransduction has been found in cardiovascular diseases such as sepsis, ischemia-reperfusion, hypertension, and diabetes. While the critical involvement of ESG in cardiovascular diseases has been established, its biomechanical properties, as well as the mechanisms underlying its normal mechanotransduction, have resisted elucidation. This lack of progress is due in large part to the fact that mechanical forces and responses that occur at the molecular and sub-cellular levels are transient, minute and therefore difficult to trace and measure. The goal of the current proposal is to develop new research tools and model systems and use them to uncover the biomechanical and mechanotranduction properties of ESG. We will characterize mechanisms underlying ESG-mediated mechanotransduction on a single-cell level using a novel Atomic Force microscopy (AFM)-fluorescence microscopy approach. The proposed study will achieve a clearer understanding of ESG-mediated mechanotransductory function, with important implications for ESG-related diseases, such as sepsis, ischemiareperfusion, diabetes and hypertension, and their therapeutics.