通过呼出气体实时监测肝衰竭的方法研究

Liver diseases are still universal in our country. Hepatic failure is a critical stage of liver diseases, with acute onset, change rapidly, clinical observation difficultly and high mortality rate (＞50%). Therefore, it is significant and urgent to find a analytic system for the real-time monitoring of patients with hepatic failure from molecular level. This project intends to use the noninvasive and ease acquisitive exhaled gas as the sample and base on the electrospray ionization mass spectrometry (EESI - MS) technique to develop a exhaled gas collecting device for the different stages of patients with liver diseases and to create a mass spectrometry technology platform for the real-time monitoring of different stages of patients with liver diseases. organic amine, nicotine, acetone, etc are adopted to simulate the exhaled gas sample to optimize the platform, thus, the real-time monitoring of exhaled gas for the different stages of patients with liver failure will be realized. Then, the spectrum of exhaled gas in the different stages of patients with liver diseases will be acquired. On this basis, combined with chemometrics methods, exploring the correlation about molecular characteristics of spectrum and different stages of disease progression or rehabilitation, then a early alarming model for the critically severe patients with liver failure will be set up. Therefore, it will dramatically improve the facility and scientificity of diagnosis in critically severe patients with liver diseases. Then improve the curative ratio of liver failure. This project firstly establish the mass spectrum analytic method which can be applied to different stages of patients with liver disease by the real-time monitoring of exhaled gas in the world, will provide new ideas for the clinical diagnosis and clinical observation of therapeutic effect on liver diseases.

我国是肝病大国。肝衰竭为肝病危重阶段，具有发病急、变化快、诊察难、死亡率高（＞50%）等特点，急需从分子层次对肝衰竭进行实时监测分析方法。本项目拟选择无创易得的呼出气体为样品，以电喷雾萃取电离质谱（EESI-MS）为基础，研制适用于采集不同阶段肝病患者呼出气体的装置，构建各阶段肝病患者呼出气体实时检测的质谱技术平台，并采用有机胺、尼古丁、丙酮等模拟呼出气体样品对平台进行优化和表征，实现对早、中、晚期肝衰竭患者呼出气体的实时质谱分析，获取肝病各阶段病人呼出气体质谱图。在此基础上，结合化学计量学方法，探究指纹谱图分子特征与不同病情发展的关联性，建立肝衰竭危重患者的预警模型，从而显著改善对肝病危重患者诊察便捷性和科学性，提高肝衰竭治愈率。本研究在国际上率先建立通过呼出气体实时监测肝衰竭的质谱分析方法，为肝脏疾病临床诊察及疗效评价提供新方法, 为其他疾病临床诊断及疗效预测提供新思路。

我国是肝病大国，肝衰竭为肝病危重阶段，具有起病急、进展快、诊察难、死亡率高等特点，现有方法尚不能满足肝衰竭诊断和病情评估要求，急需新的方法对肝衰竭进行实时监测分析。本项目从分子水平出发，利用无创易得的呼出气体为样品，以电喷雾萃取电离质谱（EESI-MS）技术为基础，研制了适用于不同阶段肝病患者的呼出气体采集装置，构建了各阶段肝病患者呼出气体实时检测的质谱技术平台，并利用丙酮及健康志愿者呼出气体对平台进行优化及表征。将该平台用于不同阶段肝病、肝衰竭患者的呼出气体实时质谱分析，结合化学计量学方法，通过构建预测模型，获得了指纹谱图分子特征与不同阶段肝病、肝衰竭以及人工肝治疗干预的关联性，结果表明，不仅肝衰竭患者与健康志愿者、慢性乙型肝炎患者的呼出气体存在显著差异，肝衰竭患者经人工肝治疗前后的呼出气体也同样存在显著差异，并进一步在各组之间筛选了多种差异性代谢物。此外，为寻求呼出气体与血液中差异性代谢物的相关性，本项目还进行了肝衰竭血液代谢组学研究，在肝衰竭患者与健康志愿者、慢性乙型肝炎患者及肝衰竭患者经人工肝治疗前后的血液中同样筛选了多种差异性代谢物。本项目研究结果为进一步开展呼出气体代谢通路相关研究及寻找肝衰竭呼出气体特异性生物标志物奠定了基础，有望为肝衰竭的临床诊察及疗效评价提供新方法。