生物标志物介导脓毒症大鼠血液净化治疗的实验研究

Sepsis is a common problem in critical care medicine, and also one of the main problems to induce the greatest expenditure of medical resources.Blood purification is potentially promising therapy for sepsis.However,the efficacy for blood purification used for sepsis is disputed, and the agreements for initial time and intensity of blood purification used for sepsis haven't been reached yet.We are the first in the world to set up the blood purification device in the rat models, and explore systemically the mechanisms of blood purification used in sepsis. We are also searching for the biomarkers to predict the development and outcome of sepsis. It becomes more and more important to guide therapies with these biomarkers, but they have never been used for blood purification.This project is to isolate the biomarkers from the blood and urine samples of septic rats using flow Luminex, ELISA, and mass-spectrometry. Then we validate and optimize the best biomarkers to predict the progress and outcome for sepsis. Finally we set up the cut-off values to initiate the starting time and intensity for blood purification, and compare the outcome between the biomarkers guided therapy and without-biomarkers guided therapy. This project will provide an ideal direction for blood purification used in sepsis.

脓毒症为重症医学科中常见病，也是消耗医疗资源最多的病种。血液净化为脓毒症的潜在治疗方向。但目前对血液净化治疗脓毒症的疗效仍有争论, 有关其适应证的选择及治疗的强度仍未达共识。我们在世界上首次建立了血液净化的大鼠模型，并对血液净化治疗脓毒症的机制进行了系统的阐述。我们最近几年对预测脓毒症发生，发展及预后的生物标志物进行了系统的筛查。标志物指导临床治疗已变得越来越重要，但这些指标对血液净化治疗脓毒症的指导意义尚未报道。本课题旨在用液相芯片分析系统，酶联免疫吸附技术，质谱仪等先进技术从脓毒症大鼠的血液、尿中分离筛选出生物标志物，寻找最佳的能预测监控脓毒症发展的标志物，选择最适浓度的标志物，并以此作为指导血液净化治疗的参照物,为进一步优化血液净化治疗脓毒症提供方向。

脓毒症为重症医学科中常见病，也是消耗医疗资源最多的病种，血液净化是脓毒症的潜在治疗方向。我们通过去除大网膜的方法改良CLP，建立了中重度肾损伤的模型，基于稳定的脓毒症肾损伤大鼠模型，我们用无标记质谱的方法检测脓毒症肾损伤大鼠尿液中的差异蛋白，分析差异蛋白所参与的生物学过程，包括免疫球蛋白的产生、损伤和防御反应，KEGG分析确定了77个显著改变的信号通路。我们进一步检测了脓毒症患者尿液中差异表达的蛋白，PARK7和CDH16的表达持续升高，并与患者尿液中Scr水平正相关。在改良的脓毒症肾损伤模型中，根据不同浓度的PARK7和CDH16进行静脉-静脉回路的大鼠血液净化模型，分析尿液PARK7和CDH16对于指导血液净化，改善肾功能和大鼠生存率的作用。综上所述，我们改良了脓毒症AKI模型，建立了稳定诱导中重度的肾损伤的CLP模型，分析了脓毒症AKI在多个时间点的蛋白组学变化，阐明了相关的生物学过程，并发现了早期诊断脓毒症肾损伤的生物标志物，为进一步研究其分子机制提供了理论基础；我们在世界上首次建立了血液净化的大鼠模型，用PARK7和CDH16两个生物标志物指导大鼠血液净化，改善肾损伤，为生物标志物指导血液净化的临床应用奠定了基础。