Front Immunol:谢青教授团队发现药物性肝损伤进展和转归相关新型代谢标志物

2022-05-21 瑞金医院 瑞金医院

2022年4月,谢青教授在Frontiers in Immunology杂志在线发表题名“Alteration of Bile Acids and Omega-6 PUFAs Are Co

2022年4月,谢青教授在Frontiers in Immunology杂志在线发表题名“Alteration of Bile Acids and Omega-6 PUFAs Are Correlated With the Progression and Prognosis of Drug Induced Liver Injury.“研究发现胆汁酸和 Omega-6 多不饱和脂肪酸与药物性肝损伤的进展和预后相关。感染科谢青主任医师、项晓刚副主任医师、王晓琳副主任医师为论文的共同通讯作者,赵爽博士研究生、付豪爽硕士研究生、周天慧博士为论文的共同第一作者。

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药物引起的肝损伤(DILI)是一个日益严重的健康问题。更重要的是,相当一部分DILI患者即使在停止使用可疑药物后,肝功能仍然不能完全恢复。慢性DILI表现为持续或反复发作的炎症或胆管的减少,甚至进展到肝硬化,最终需要肝移植,因此是一个亟待解决的临床问题。然而,迄今为止对慢性DILI的研究仍然十分匮乏,包括DILI慢性化机制、危险因素和治疗策略等,也没有广泛认可的相关生物标志物。肝脏是人体的主要代谢中心,探索DILI的代谢特征,有望发现与疾病进展及预后相关的新的生物标志物。

本研究招募了 119 名 DILI 患者和 156 名健康对照 (HC)。 最终分析共纳入 90 名住院 DILI 患者和 70 名 HC进行了分析,示意图如下。

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在我们的研究中,包括 90 名 DILI 患者和 70 名 HC。 DILI患者根据结果分为两组:恢复的DILI(DILI.rec)和慢性DILI(DILI.chr)组。 根据DILI严重程度的标准(22),将患者分为轻度(1级)、中度(2级)、重度(3级)和肝功能衰竭(4级)。 此外,我们将 1 级和 2 级定义为非关键组(DILI.a),将 3 级和 4 级定义为关键组(DILI.b)。

本文通过超高效液相色谱-串联质谱对DILI患者和健康对照者血浆中脂肪酸、氨基酸、有机酸、碳水化合物和胆汁酸等进行了全面且定量的研究。 

通常,PUFA 包括 omega-6 和 omega-3 PUFA。 Omega-3 PUFAs主要包括发挥抗炎作用的ALA、二十碳五烯酸(EPA)和DHA,而omega-6 PUFAs主要包括促炎作用的LA、GLA、花生四烯酸(AA)、AdA和DPAn-6 (23, 24)。在 DILI.rec 和 DILI.chr 组之间比较了 PUFA 的 omega-6 和 omega-3 成分的水平。 LA/ALA 比值无显着差异,而 (AdA+GLA+LA)/(EPA+DHA) 比值显着增加(DILI.chr vs DILI.rec 组)(图 4F)。丙二醛 (MDA) 是 PUFA 的一种代谢终产物,可被 DILI 中的氧化应激过氧化,已被广泛用作脂质过氧化的标志物。 GSH-Px 是保护细胞免受活性氧 (ROS) 增加的主要抗氧化成分 (27)。与 HCs 相比,DILI 患者的血清 MDA 或 MDA/GSH-Px 水平上调,与 DILI.rec 组相比,DILI.chr 组略高(图 4G)。 AA、AdA水平或(AdA+GLA+LA)/(EPA+DHA)比值与MDA和MDA/GSH-PX呈正相关(图4H)。

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进一步通过代谢组分析发现,胆汁酸代谢物甘氨鹅脱氧胆酸(GCDCA)、甘氨胆酸(GCA)、牛磺鹅去氧胆酸(TCDCA)、牛磺胆酸(TCA)是区分DILI严重程度的有效标志,而DILI中高水平的多不饱和脂肪酸(PUFA)尤其是n6-PUFAs与DILI慢性化转归密切相关。花生四烯酸和肾上腺酸与过氧化指标MDA和MDA/GSH-Px正相关。编码脂质过氧化相关酶的 ALOX5、ALOX12 和 COX-1在DILI患者的肝脏中高表达,提示PUFA过氧化可能导致DILI慢性化转归有关。

进一步,研究者开发一个模型来预测哪些患者能转变为慢性 DILI 。采用随机森林方法处理高维数据,用于选择代谢物以区分慢性(DILI.chr)和恢复期(DILI.rec)(GINI>1)。选定的代谢物连同直接胆红素 (DB) 和总胆红素 (TB) (t 检验 p < 0.05) 被确定为预测模型的候选变量。然后,我们将 DILI 患者随机分为训练集(n=60)和验证集(n=30),其中康复患者和慢性患者数量相匹配。在训练集中进行的单变量logistic回归将 DPA、天冬氨酸、肾上腺酸、酪氨酸、GLA、富马酸、酪氨酸、琥珀酸和乳酸确定为预测 DILI 慢性病风险的独立因素(表 2)。随后,采用二元logistic回归分析和在训练集中进行优化算法来消除多重共线性,并从这些独立的风险因素中选择最佳变量。最后,AdA 和 Asp 的组合(“肾上腺酸-天冬氨酸”)被定义为基于训练集的理想面板(表 2)。

恢复期 DILI 与慢性 DILI 训练集的 ROC 曲线下面积(AUC)为 0.889,敏感性为 73.7%,特异性为 92.7%,而验证集的 AUC 为 0.888,灵敏度为 71.4%,特异性为 91.3%(图 5B)。然后,研究者在所有 DILI 患者中评估了 AdA 和 Asp(“AdA-Asp 模型”)与其他标志物(如 TB、DB、ALP 和 TCA)的预测能力,TB、DB、ALP 和 TCA 的 AUC 分别为 0.57、0.55、0.58 和 0.57,而 AdA 和 Asp 组的 AUC 为 0.850(图 5C)。列线图预测与实际观察之间的最佳一致性(图 5D)。在决策曲线分析中,AdA-Asp 模型在合理的阈值范围内提供了相当大的净收益(图 5E),表明该模型具有广阔的临床应用前景。    

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进一步研究表明,在疾病的急性期,肠道微生物群可能分别通过FGF19信号通路和促进PUFA饱和在维持胆汁酸和PUFAs的稳态中发挥有益作用,尽管这种负反馈并不能完全抵消PUFAs和胆汁酸的累积。  

本研究填补了DILI相关领域的空白,第一次基于广谱的代谢组学研究明确提出了多不饱和脂肪酸是与DILI慢性化转归最密切相关的代谢组分。基于关键代谢物构建的临床预测模型,有利于临床早期识别有慢性化风险的DILI患者,进而获得早期干预的机会。本研究也加深了我们对DILI进展和慢性化机制的理解,并表明代谢物和肠道微生物群可能是DILI临床干预的新靶点。

原始出处:

Zhao S, Fu H, Zhou T, Cai M, Huang Y, Gan Q, Zhang C, Qian C, Wang J, Zhang Z, Wang X, Xiang X, Xie Q.Alteration of Bile Acids and Omega-6 PUFAs Are Correlated With the Progression and Prognosis of Drug-Induced Liver Injury.Front Immunol. 2022 Apr 12;13:772368. doi: 10.3389/fimmu.2022.772368

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    2022-05-21 病毒猎手

    #代谢组学#价值大!

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