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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(P<0.01),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(P>0.05)。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。
Objective To analyze the differences of bile acid sub-components in patients with cirrhosis and healthy people and to explore its value in the diagnosis of cirrhosis. Methods According to the diagnostic criteria, patients with liver cirrhosis who were hospitalized in Shanghai Tenth People’s Hospital from 2016 to 2017 and healthy people in our hospital during the same period were selected as research objects. The levels of serum bile acid in the two groups were quantitatively determined by high performance liquid chromatography-mass spectrometry The data were analyzed by principal component analysis (PCA), and the data were analyzed by Mann-Whitney test. Results There were 5 kinds of free bile acids (BAs), 5 kinds of taurine-binding bile acids (T-BAs) and 5 kinds of glycine-conjugated bile acids (G-BAs) . Compared with the healthy control group, 12 metabolites in serum of cirrhosis group were significantly different (P <0.01), including all primary bile acids synthesized in the liver and some secondary bile acids synthesized in the intestine Cholic acid, glycochloridic acid and taurolithocholic acid), of which taurocholic acid and glycocholic acid increased significantly, while lithocholic acid, glycodeoxycholic acid and taurodeoxycholic acid showed no significant difference Statistical significance (P> 0.05). PCA results showed that the two groups of people in the PC1 area is larger. Conclusion The established LC-MS / MS method for simultaneous determination of 15 bile acid components in trace serum is stable and reliable, which can meet the needs of clinical quantitative detection. The serum biliary acid abnormalities detected initially by serum of patients with cirrhosis are suggested. The change of bile acid component is closely related to the anomalous function of liver anabolism and metabolism, which provides some research bases for studying the role of bile acid in the development of liver cirrhosis.