基于微流控芯片的糖尿病患者中性粒细胞的趋化性和吞噬功能研究

The immune function of diabetic patients is closely related to the dysfunction of chemotaxis and phagocytosis of neutrophils. Studying the chemotaxis and whole phagocytosis process of their neutrophils is very helpful to evaluate the immune function and therapeutic effect of diabetic patients. In this project, a novel microfluidic device capable of rapid separation of neutrophils is developed and directly used for the assessment of neutrophils’ chemotaxis and phagocytosis. Then, neutrophil phenotype-related parameters, including the changes in pseudarthrosis elongation and retraction, adhesion index, chemotactic index, migration speed, pathogen binding index and phagocytic index, and the difference between healthy and diabetic patients, are analyzed and explored. In addition, the relationship between neutrophil function-related parameters and conventional immune functional factors are studied. Finally, statistical methods were used to analyze the specific contributions of various cell functional parameters in the differentiation of diabetic and healthy group. Combining above results, the neutrophil test of well-controlled diabetic patients is carried out to evaluate the application value of neutrophil chemotaxis and phagocytic function in monitoring the immune function and prognosis of diabetes. These results are expected to establish an analytical method based on microfluidic device for investigating the neutrophil chemotaxis and phagocytosis, thus providing reference data and basic case to evaluate the immune function and prognosis of diabetic patients in terms of cellular function phenotype.

糖尿病患者易并发感染与中性粒细胞的趋化性和吞噬功能障碍密切相关。研究中性粒细胞的趋化运动及其发挥吞噬作用的全过程，有助于评估糖尿病患者的免疫功能状态和治疗效果。本项目拟首先设计集成中性粒细胞快速分离、趋化性和吞噬功能检测为一体的微流控芯片；接着，重点分析糖尿病人和正常人中性粒细胞的趋化性和吞噬功能各个评价参数（伪足伸展变化规律、粘附性指数、趋化性指数、趋化运动速度、病原体结合指数和吞噬指数）及其差异性，并探究细胞功能参数及其常规免疫功能指标之间的相关性；最后，采用统计学方法分析各个细胞功能参数在区分糖尿病组和正常人群中的具体贡献，并结合病情控制良好糖尿病组实验，探究中性粒细胞的趋化性和吞噬功能在监测糖尿病免疫健康状态和预后方面的应用价值。项目预期成果有望建立基于微流控芯片的中性粒细胞趋化性和吞噬功能分析方法，并为从细胞功能表型方面评价糖尿病患者的免疫功能和预后提供参考数据和基本案例。

采用微流控芯片研究糖尿病患者中性粒细胞的趋化性和吞噬功能，对于评估糖尿病患者的免疫健康状态和进行疗效评估具有重要的科学意义。主要研究内容：①完成了集成细胞定位功能的多通道微流控芯片关键技术研究；②完成了细胞快速分选和便携式分析装置关键技术研究；③建立了微流控芯片上糖尿病患者中性粒细胞的趋化性和吞噬功能快速评价方法和参数体系。研究结果：①与正常人相比，糖尿病病人除糖化血红蛋白水平异常升高之外，中性粒细胞的趋化功能明显受损。②依据糖化血红蛋白浓度对糖尿病人进行风险分层，发现“高风险患者”的均值CRP、总胆固醇水平高于“低风险者”，“高风险患者”的平均运动速度高于低风险者。③糖尿病人中的中性粒细胞吞噬功能明显受损。研究成果意义：①为相关科研人员开展研究提供了理论基础和技术储备。②提示微流控芯片的细胞趋化性和吞噬功能有望成为进行糖尿病等慢病患者的炎症风险评估的新型生物标志物。