Table 1_Evaluation of the safety and efficacy of Sophorae Flavescentis Radix extract in the treatment of inflammatory bowel disease based on zebrafish models.docx

BackgroundSophorae Flavescentis Radix (Kushen) has been employed in Traditional Chinese Medicine (TCM) for over 2,000 years, primarily for its effects in clearing heat and drying dampness, eliminating parasites, and promoting diuresis. Preliminary findings suggest that Kushen extract or its formulations may exhibit potential therapeutic benefits in alleviating inflammatory bowel disease (IBD).
AimsThis study aimed to comprehensively evaluate the safety and efficacy of Kushen extract using a zebrafish model, with a particular focus on intestinal mucosal immunity, and to explore its underlying mechanisms.
Material and methodsThe safety and efficacy profile of Kushen extract was comprehensively evaluated in zebrafish model. Network pharmacology combined with transcriptomics were employed to predict the mechanisms underlying Kushen extract’s therapeutic effects on IBD, followed by validation using Reverse Transcription-Quantitative Polymerase Chain Reaction (RT-qPCR) and molecular docking.
ResultsAdministration of supra-therapeutic doses of Kushen extract induced hepatotoxicity effects in zebrafish, primarily manifested as hepatic morphological abnormalities and exacerbated hepatocyte apoptosis. Conversely, low-dose administration of Kushen extract demonstrated significant therapeutic benefits including improved intestinal phagocytic function, reduced gut neutrophil infiltration and enhanced goblet cell secretion. Furthermore, Kushen extract treatment at low doses significantly decreased the levels of pro-inflammatory cytokines. Integrated network pharmacology and transcriptomic analyses indicated that the amelioration of Kushen extract on IBD may be involved in FoxO4/NOD-like/Apoptosis and MAPK signaling pathways. The RT-qPCR results confirmed that Kushen extract effectively modulates the expression levels of key genes in the aforementioned pathway. Molecular docking analysis revealed binding energies of -8.1 kcal/mol between matrine and the BCL-2 protein, and -10.4 kcal/mol between oxymatrine and EGFR, indicating strong molecular interactions between these compounds and their target proteins. These interactions may collectively contribute to the therapeutic efficacy of Kushen extract.
ConclusionsSupra-therapeutic doses of Kushen extract can induce marked liver damage in zebrafish. However, when administration within an appropriate dosage range, it does not elicit significant toxicity and demonstrates substantial therapeutic efficacy against IBD, partly through modulating mucosal immunity. These findings provide crucial experimental evidence to support the guidance for safe and rational clinical application of Kushen extract.

背景：苦参（Sophorae Flavescentis Radix，Kushen）在中医药（Traditional Chinese Medicine, TCM）中应用已有两千余年历史，主要功效为清热燥湿、杀虫、利尿。前期研究表明，苦参提取物或其制剂在缓解炎症性肠病（Inflammatory Bowel Disease, IBD）方面可能具有潜在治疗价值。

研究目的：本研究旨在利用斑马鱼模型全面评估苦参提取物的安全性与有效性，重点关注肠黏膜免疫，并探索其潜在作用机制。

材料与方法：本研究通过斑马鱼模型全面评价苦参提取物的安全性与有效性谱。采用网络药理学结合转录组学技术，预测苦参提取物治疗炎症性肠病的潜在作用机制，随后通过逆转录定量聚合酶链反应（Reverse Transcription-Quantitative Polymerase Chain Reaction, RT-qPCR）与分子对接技术进行验证。

结果：给予超治疗剂量的苦参提取物可诱导斑马鱼产生肝毒性，主要表现为肝脏形态异常及肝细胞凋亡加剧。与之相反，低剂量苦参提取物则展现出显著的治疗益处：包括改善肠道吞噬功能、减少肠道中性粒细胞浸润、增强杯状细胞分泌功能。此外，低剂量苦参提取物可显著降低促炎细胞因子水平。整合网络药理学与转录组学分析显示，苦参提取物对炎症性肠病的改善作用可能与FoxO4/NOD样/凋亡及MAPK信号通路相关。RT-qPCR结果证实，苦参提取物可有效调控上述通路中的关键基因表达水平。分子对接分析显示，苦参碱（matrine）与BCL-2蛋白的结合能为-8.1 kcal/mol，氧化苦参碱（oxymatrine）与EGFR的结合能为-10.4 kcal/mol，表明这些化合物与其靶蛋白之间存在较强的分子相互作用，此类相互作用可能共同促成苦参提取物的治疗功效。

结论：超治疗剂量的苦参提取物可在斑马鱼体内引发明显肝损伤。然而，当给药剂量处于适宜范围时，苦参提取物不会引发显著毒性，且对炎症性肠病展现出可观的治疗功效，其部分作用机制为调控黏膜免疫。本研究结果为苦参提取物安全合理的临床应用提供了重要实验依据。