Molecular mechanisms involved in drug-induced liver injury caused by urate-lowering Chinese herbs: A network pharmacology study and biology experiments

As an important part of the comprehensive treatment methods, the urate-lowering Chinese herbs could provide favorable clinical effects on hyperuricemia in its ability to invigorate spleen and remove dampness. Owing to the long-term duration, it brought up the potential adverse reactions (ADRs) and concerns about the drug-induced liver injury from these herbs. To address this problem, the bioinformatics approaches which combined the network pharmacology, computer simulation and molecular biology experiments were undertaken to elucidate the underlying drug-induced liver injury molecular mechanisms of urate-lowering Chinese herbs. Several electronic databases were searched to identify the potential liver injury compounds in published research. Then, the putative target profile of liver injury was predicted, and the interaction network was constructed based on the links between the compounds, corresponding targets and core pathways. Accordingly, the molecular docking simulation was performed to recognize the representative compounds with hepatotoxicity. Finally, the cell experiments were conducted to investigate the biochemical indicators and expression of the crucial protein that were closely associated with liver injury. In conclusion, the current research revealed that the compounds with potential liver injury including diosgenin, baicalin, saikosaponin D, tetrandrine, rutaecarpine and evodiamine from urate-lowering Chinese herbs, could lead to decline the survival rate of L-02 cell, increase the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) in cell-culture medium, enhance the expression of p-p38/p38, while the p38 inhibitor could achieve the trend of regulating and controlling liver injury. These research findings bring further support to the growing evidence that the mechanism of the liver injury induced by the compounds from urate-lowering Chinese herbs may be associated with the activation of p38α.

作为综合治疗方案的重要组成部分，降尿酸中草药（urate-lowering Chinese herbs）凭借健脾祛湿的功效，可对高尿酸血症（hyperuricemia）展现出良好的临床疗效。由于需长期用药，该类药物引发的潜在不良反应（ADRs）以及药物性肝损伤（drug-induced liver injury）相关担忧也逐渐显现。为解决这一问题，本研究采用整合网络药理学（network pharmacology）、计算机模拟与分子生物学实验的生物信息学方法，旨在阐明降尿酸中草药潜在的药物性肝损伤分子机制。研究团队通过检索多个电子数据库，从已发表的研究中筛选出潜在的肝损伤相关化合物；随后预测肝损伤的潜在靶点谱，并基于化合物、对应靶点与核心通路间的关联构建相互作用网络。据此开展分子对接模拟（molecular docking simulation），以筛选出具有肝毒性的代表性化合物。最后通过细胞实验，检测与肝损伤密切相关的生化指标及关键蛋白的表达水平。本研究最终发现，降尿酸中草药中含有的薯蓣皂苷元（diosgenin）、黄芩苷（baicalin）、柴胡皂苷D（saikosaponin D）、粉防己碱（tetrandrine）、吴茱萸碱（rutaecarpine）与吴茱萸次碱（evodiamine）等潜在肝损伤化合物，可降低L-02细胞的存活率，升高细胞培养液（cell-culture medium）中天冬氨酸氨基转移酶（aspartate aminotransferase, AST）、丙氨酸氨基转移酶（alanine aminotransferase, ALT）、乳酸脱氢酶（lactate dehydrogenase, LDH）与碱性磷酸酶（alkaline phosphatase, ALP）的活性，增强p-p38/p38的表达；而p38抑制剂可逆转该肝损伤的发生趋势。上述研究结果进一步佐证了日益增多的证据，即降尿酸中草药来源的化合物诱导肝损伤的机制可能与p38α的激活相关。