肝硬化患者血清胆汁酸代谢物谱的检测分析

目的分析肝硬化患者与健康体检人群微量血清中胆汁酸亚组分含量差异,并探讨其在肝硬化患者诊断中的价值。方法选取符合诊断标准的2016—2017年在上海第十人民医院住院的肝硬化患者和同期在本院健康体检人群作为研究对象,利用高效液相色谱-质谱法定量检测两组人群血清胆汁酸组分含量,应用主成分分析法(PCA)对数据进行代谢轮廓识别,并采用Mann-Whitney检验方法对定量数据进行统计分析。结果该检测体系中,两组人群血清样本共检测得到5种游离胆汁酸(BAs)、5种牛磺结合型胆汁酸(T-BAs)和5种甘氨结合型胆汁酸(G-BAs)。相比于健康对照组,肝硬化组血清中12个代谢物差异有统计学意义(P0.01),包括在肝脏内合成的所有初级胆汁酸和在肠道中合成的部分次级胆汁酸(脱氧胆酸、甘氨石胆酸和牛磺石胆酸),其中以牛磺胆酸和甘氨胆酸升高幅度较大,而石胆酸、甘氨脱氧胆酸和牛磺脱氧胆酸比较差异无统计学意义(P0.05)。PCA结果显示两组人群在PC1区区分较大。结论所建立的在微量血清中同时检测15种胆汁酸组分的LC-MS/MS方法稳定可靠,可满足临床定量检测需求;利用该方法检测肝硬化患者血清初步发现胆汁酸代谢谱异常,提示胆汁酸组分的改变与肝脏合成代谢功能异常密切相关,为研究胆汁酸在肝硬化发展进程中的作用提供了一定的研究基础。

Detection and analysis of serum bile acid metabolism in liver cirrhosis patients

Objective To analyze the difference between the liver cirrhosis patients and health subjects in their microserum bile acid sub-components,and explore the diagnostic value of liver cirrhosis.Methods The inpatients with liver cirrhosis in accordance with the diagnostic criteria and health examination population were selected as research subjects from 2016 to 2017.The high performance liquid chromatography-mass spectrometry (LC-MS/MS) method was used to quantitatively detect the levels of serum bile acid in the two groups.The principal component analysis (PCA) was used to identify the metabolic profile of data,and the Mann-Whitney testing method was used to statistically analyze the quantitative data.Results Five kinds of free bile acids(BAs),five kinds of taurine-binding bile acids(T-BAs) and five kinds of glycine-binding bile acids(G-BAs) were detected in the serum samples of the two groups in this test system.There were significant differences in 12 metabolites between the two groups(P＜0.01),including all primary bile acids synthesized in the liver and part of the secondary bile acids synthesized in the gut(DCA,GLCA and TLCA),and TCA and GCA increased a larger range among them in the liver cirrhosis group.LCA,GDCA and TDCA showed no significant difference between the two groups (P＞0.05).PCA results showed that the two groups were largely distinguished in PC1.Conclusion The LC-MS/MS method for the simultaneous detection of 15 bile acid components in micro-serum is stable and reliable,which could meet the needs of clinical quantitative detection.The application of the method for testing serum bile acid in patients with liver cirrhosis can preliminary find metabolism spectrum anomaly of bile acid,suggesting that the changes of bile acid components are closely related with the dysfunction of liver synthesis and metabolism,which may provide the certain research foundation for studying the function of bile acid in the development process of cirrhosis.