代谢组学在肝脏疾病研究中的应用进展

代谢组学是通过考察生物体受到来自体内外的各种刺激或扰动时内源性小分子代谢物所发生的变化,来阐明生物体系的代谢途径的一种较新的组学技术.肝脏作为生物体最大、最复杂的代谢器官,参与了体内绝大部分的代谢过程.肝脏病变时,其相关的代谢网络必然会发生相应的变化.近几年,代谢组学技术在肝脏疾病研究中的应用日益增多,此文对代谢组学的概念、研究现状、研究方法及在肝脏疾病的基础和临床研究中的应用进行综述.     

A serum metabonomics study on the difference between alcohol- and HBV-induced liver cirrhosis by UPLC coupled with Q-TOF mass spectrometry

Background Liver cirrhosis is the fatal sequel of chronic hepatitis, making early diagnosis of liver cirrhosis critical. Liver biopsy is still the standard diagnostic method for liver cirrhosis, although its use in a wide population with alcoholism or HBV infection remains difficult. In this study, we used the metabonomic approach to detect potential biomarkers for early diagnosis of liver cirrhosis. Methods Serums were collected prospectively from normal control subjects (n=22) and patients with alcoholic cirrhosis (n=18) or hepatitis B virus (HBV)-induced cirrhosis (n=19). The serum metabonome was analyzed using ultraperformance liquid chromatography (LC)/time-of-flight mass spectrometry (MS) integrated with chemometrics. The acquired LC-MS data were normalized and processed using principal component analysis and partial least squares discrimination analysis. Results Significant differences in the metabonome among the three groups were observed. Lysophosphatidyl cholines (LPCs) (LPC C16:0, LPC C18:0, LPC C18:2, LPC C18:3, LPC C20:3, LPC C20:5) decreased in the serum of patients with hepatic cirrhosis, whereas bile acids (glycocholic acid, glycochenodeoxycholic acid), hypoxanthine, and stearamide increased in the serum of patients with liver cirrhosis. These metabolites are considered “common” biomarkers for hepatic cirrhosis. Oleamide and myristamide increased in the serum of patients with alcoholic cirrhosis but decreased in those with HBV-induced cirrhosis. These could be specific biomarkers for differential diagnosis between alcohol- and HBV-induced hepatic cirrhosis. Conclusions There are significance metabonomic differences between alcohol- and HBV-induced liver cirrhosis. This study demonstrates that metabonomics is a top-down system biology tool for conducting research on clinical problems.     

非生物人工肝支持系统治疗肝功能衰竭的血清氨基酸谱变化

Objective To analyze the patterns of amino acid changes in liver failure patients treated with non-bioartificial liver support system (ALSS), and to explore the efficacy of ALSS in liver failure treatment. Methods A total of 146 liver failure patients treated with ALSS from June 2009 to August 2010 were recruited in this study. Paired blood samples were collected from every patient and serum amino acids and ammonia were tested by automatic amino acid analyzer. The changes of amino acids in patients with different prognoses, different types/phases of liver failure were evaluated.Measurement data were compared by paired t test. Results After ALSS treatment, liver failure patients experienced a significant decrease in serum glutamic acid and lysine [(395.62±200.24)μmol/Lvs (260. 05±169.56) μmol/L and (436. 73±326. 18)μmol/L vs (407. 12±292.01) μmol/L,respectively; t= 8. 611 and 2. 659, respectively; both P＜0.01)], while experienced greatly increases in threonine and branched-chain amino acids/aromatic amino acid ratio [( 1302. 90 ±1288.70) μmol/L vs (1406.70 ±1272. 34) μmol/L and 1. 23 ± 0. 53 vs 1. 36 ± 0.57, respectively; t = 2. 895 and 1. 061,respectively; both P＜0. 01)]. The changes of glutamic acid, tyrosine, arginine and methionine before and after ALSS treatment in patients with different prognoses, different types/phases of liver failure were all significantly different. Conclusions ALSS treatment could improve the serum amino acid disorder in liver failure patients. The amino acids in patients with different types/phases or different prognoses of liver failure change significantly after ALSS treatment.     

非生物人工肝支持系统治疗肝功能衰竭的血清氨基酸谱变化

目的 分析肝功能衰竭患者接受非生物人工肝支持系统(ALSS)治疗引起的血清氨基酸谱变化,阐述非生物ALSS治疗肝功能衰竭的有效性.方法 对2009年6月至2010年8月间146例肝功能衰竭患者行非生物ALSS治疗,观察治疗前后患者血清氨基酸谱的变化情况,并对不同预后、不同肝功能衰竭类型以及处于不同肝功能衰竭时期患者的血清氨基酸水平的变化进行研究.计量资料采用配对样本的t检验.结果 非生物ALSS治疗后,血清谷氨酸和赖氨酸下降明显[(395.62±200.24)μumol/L比(260.05±169.56)μmol/L,(436.73±326.18)μmol/L比(407.12±292.01)μmol/L;t＝8.611,t＝2.659;均P＜0.01],苏氨酸、支链氨基酸/芳香族氨基酸显著上升[(1302.90±1288.70)μmol/L比(1406.70±1272.34)μmol/L,1.23±0.53比1.36±0.57;t＝2.895,t＝1.061;均P＜0.01].在预后不同的患者间、不同肝功能衰竭类型以及处于不同时期的肝功能衰竭患者间,谷氨酸、酪氨酸、精氨酸以及蛋氨酸等治疗前后差异有统计学意义.结论 非生物ALSS治疗可以改善肝功能衰竭患者血清氨基酸紊乱状况.不同肝功能衰竭分类、分期以及不同预后患者的血清氨基酸谱在非生物ALSS治疗后有显著不同的变化.

Abstract：

Objective To analyze the patterns of amino acid changes in liver failure patients treated with non-bioartificial liver support system (ALSS), and to explore the efficacy of ALSS in liver failure treatment. Methods A total of 146 liver failure patients treated with ALSS from June 2009 to August 2010 were recruited in this study. Paired blood samples were collected from every patient and serum amino acids and ammonia were tested by automatic amino acid analyzer. The changes of amino acids in patients with different prognoses, different types/phases of liver failure were evaluated.Measurement data were compared by paired t test. Results After ALSS treatment, liver failure patients experienced a significant decrease in serum glutamic acid and lysine [(395.62±200.24)μmol/Lvs (260. 05±169.56) μmol/L and (436. 73±326. 18)μmol/L vs (407. 12±292.01) μmol/L,respectively; t= 8. 611 and 2. 659, respectively; both P＜0.01)], while experienced greatly increases in threonine and branched-chain amino acids/aromatic amino acid ratio [( 1302. 90 ±1288.70) μmol/L vs (1406.70 ±1272. 34) μmol/L and 1. 23 ± 0. 53 vs 1. 36 ± 0.57, respectively; t = 2. 895 and 1. 061,respectively; both P＜0. 01)]. The changes of glutamic acid, tyrosine, arginine and methionine before and after ALSS treatment in patients with different prognoses, different types/phases of liver failure were all significantly different. Conclusions ALSS treatment could improve the serum amino acid disorder in liver failure patients. The amino acids in patients with different types/phases or different prognoses of liver failure change significantly after ALSS treatment.     

非生物人工肝支持系统治疗肝功能衰竭的血清氨基酸谱变化

Objective To analyze the patterns of amino acid changes in liver failure patients treated with non-bioartificial liver support system (ALSS), and to explore the efficacy of ALSS in liver failure treatment. Methods A total of 146 liver failure patients treated with ALSS from June 2009 to August 2010 were recruited in this study. Paired blood samples were collected from every patient and serum amino acids and ammonia were tested by automatic amino acid analyzer. The changes of amino acids in patients with different prognoses, different types/phases of liver failure were evaluated.Measurement data were compared by paired t test. Results After ALSS treatment, liver failure patients experienced a significant decrease in serum glutamic acid and lysine [(395.62±200.24)μmol/Lvs (260. 05±169.56) μmol/L and (436. 73±326. 18)μmol/L vs (407. 12±292.01) μmol/L,respectively; t= 8. 611 and 2. 659, respectively; both P＜0.01)], while experienced greatly increases in threonine and branched-chain amino acids/aromatic amino acid ratio [( 1302. 90 ±1288.70) μmol/L vs (1406.70 ±1272. 34) μmol/L and 1. 23 ± 0. 53 vs 1. 36 ± 0.57, respectively; t = 2. 895 and 1. 061,respectively; both P＜0. 01)]. The changes of glutamic acid, tyrosine, arginine and methionine before and after ALSS treatment in patients with different prognoses, different types/phases of liver failure were all significantly different. Conclusions ALSS treatment could improve the serum amino acid disorder in liver failure patients. The amino acids in patients with different types/phases or different prognoses of liver failure change significantly after ALSS treatment.     

中低剂量糖皮质激素影响新型冠状病毒清除时间的回顾性研究

目的研究中低剂量糖皮质激素治疗对新型冠状病毒肺炎(COVID-19)患者病毒清除时间的影响。方法回顾性分析2020年1月19日至2月17日浙江大学医学院附属第一医院收治的72例COVID-19患者的临床资料。全部患者均接受阿比多尔和/或联合洛匹那韦/利托那韦、达芦那韦/考比司他抗病毒以及对症支持治疗,其中51例使用甲泼尼龙(0.75~1.50 mg·kg-1·d-1)抗炎治疗作为糖皮质激素治疗组,21例未使用糖皮质激素者作为对照组。比较两组患者及普通型患者痰液病毒核酸转阴时间和肺部影像学改善时间。组间比较采用Kruskal-Wallis检验或Fisher精确检验。结果糖皮质激素治疗组与对照组中位年龄分别为52(45,62)和46(32,56)岁,两组比较差异有统计学意义(χ2=4.365,P<0.05)。两组临床分型差异有统计学意义(P<0.01),糖皮质激素治疗组中重型患者占52.0%,而对照组中普通型占71.4%。两组患者从发病至痰液病毒核酸转阴中位时间分别为15(13,20)和14(12,20)d,差异无统计学意义(P>0.05);两组患者从发病至肺部影像学好转中位时间分别为13(11,15)和13(12,17)d,差异无统计学意义(P>0.05)。在普通型患者中,糖皮质激素治疗组与对照组从发病至痰液病毒核酸转阴中位时间为13(11,18)和13(12,15)d,差异无统计学意义(P>0.05);从发病至肺影像学好转中位时间分别为12(10,15)和13(12,17)d,差异也无统计学意义(P>0.05)。结论在不同临床类型的COVID-19患者中,中低剂量糖皮质激素使用对病毒清除时间未见明显影响;普通型患者中未观察到激素可以加快肺损伤的改善,故不建议使用。