State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine , Zhejiang University , Hangzhou 310003 , China.
Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases , Zhejiang University , Hangzhou 310003 , China.
J Proteome Res. 2019 Jun 7;18(6):2514-2524. doi: 10.1021/acs.jproteome.9b00047. Epub 2019 Apr 30.
To evaluate the levels of metabolites and cytokines in the serum of patients with severe and non-severe idiosyncratic drug-induced liver injury (DILI) and to identify biomarkers of DILI severity.
Gas chromatography-mass spectrometry (GC-MS) and ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) based metabolomic approaches were used to evaluate the metabolome of serum samples from 29 DILI patients of severity grade 3 (non-severe), 27 of severity grade 4 (severe), and 36 healthy control (HC). The levels of total keratin-18 (K18), fragment K18, and 27 cytokines were determined by enzyme-linked immunosorbent assay.
The alkaline phosphatase activity ( p = 0.021) and international normalized ratio (INR) ( p < 0.001) differed significantly between the severe and non-severe groups. The severe group had a higher serum fragment K18 level than the non-severe group. A multivariate analysis showed good separation between all pairs of the HC, non-severe, and severe groups. According to the orthogonal partial least-squares-discriminant analysis (OPLS-DA) model, 14 metabolites were selected by GC-MS and 17 by UPLC-MS. Among these metabolites, the levels of 16 were increased and of 15 were decreased in the severe group. A pathway analysis revealed major changes in the primary bile acid biosynthesis and alpha-linolenic acid metabolic pathways. The levels of PDGF-bb, IP-10, IL-1Rα, MIP-1β, and TNF-α differed significantly between the severe and non-severe groups, and the levels of most of the metabolites were negatively correlated with those of these cytokines. An OPLS-DA model that included the detected metabolites and cytokines revealed clear separation of the severe and non-severe groups.
We identified 31 metabolites and 5 cytokines related to the severity of idiosyncratic DILI. The primary bile acid biosynthesis and alpha-linolenic acid metabolism pathways were also related to the severity of DILI. A model that incorporated the metabolites and cytokines showed clear separation between patients with severe and non-severe DILI, suggesting that these biomarkers have potential as indicators of DILI severity.
评估严重和非严重特异质药物性肝损伤（DILI）患者血清中的代谢物和细胞因子水平，并确定 DILI 严重程度的生物标志物。
采用气相色谱-质谱（GC-MS）和超高效液相色谱-质谱（UPLC-MS）基于代谢组学的方法评估 29 例 DILI 严重程度 3 级（非严重）、27 例严重程度 4 级（严重）和 36 例健康对照（HC）患者血清样本的代谢组。通过酶联免疫吸附试验测定总角蛋白-18（K18）、片段 K18 和 27 种细胞因子的水平。
碱性磷酸酶活性（p=0.021）和国际标准化比值（INR）（p<0.001）在严重组和非严重组之间有显著差异。严重组血清片段 K18 水平高于非严重组。多元分析显示 HC、非严重和严重组之间的良好分离。根据正交偏最小二乘判别分析（OPLS-DA）模型，GC-MS 选择了 14 种代谢物，UPLC-MS 选择了 17 种代谢物。在这些代谢物中，16 种代谢物水平升高，15 种代谢物水平降低。途径分析显示主要胆汁酸生物合成和α-亚麻酸代谢途径发生了重大变化。PDGF-bb、IP-10、IL-1Rα、MIP-1β和 TNF-α在严重组和非严重组之间有显著差异，大多数代谢物的水平与这些细胞因子的水平呈负相关。包含检测到的代谢物和细胞因子的 OPLS-DA 模型清楚地区分了严重组和非严重组。
我们确定了 31 种与特异质 DILI 严重程度相关的代谢物和 5 种细胞因子。主要胆汁酸生物合成和α-亚麻酸代谢途径也与 DILI 的严重程度有关。纳入代谢物和细胞因子的模型清楚地区分了严重和非严重 DILI 患者，表明这些生物标志物具有作为 DILI 严重程度指标的潜力。
