Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, 350108, China.
Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, 350108, China; Department of Preventive Medicine, School of Public Health, Fujian Medical University, Fuzhou, 350108, China.
Chem Biol Interact. 2024 May 25;395:111036. doi: 10.1016/j.cbi.2024.111036. Epub 2024 May 4.
Gelsemium elegans Benth. (G. elegans) is a traditional medicinal herb that has anti-inflammatory, analgesic, sedative, and detumescence effects. However, it can also cause intestinal side effects such as abdominal pain and diarrhea. The toxicological mechanisms of gelsenicine are still unclear. The objective of this study was to assess enterotoxicity induced by gelsenicine in the nematodes Caenorhabditis elegans (C. elegans). The nematodes were treated with gelsenicine, and subsequently their growth, development, and locomotion behavior were evaluated. The targets of gelsenicine were predicted using PharmMapper. mRNA-seq was performed to verify the predicted targets. Intestinal permeability, ROS generation, and lipofuscin accumulation were measured. Additionally, the fluorescence intensities of GFP-labeled proteins involved in oxidative stress and unfolded protein response in endoplasmic reticulum (UPR) were quantified. As a result, the treatment of gelsenicine resulted in the inhibition of nematode lifespan, as well as reductions in body length, width, and locomotion behavior. A total of 221 targets were predicted by PharmMapper, and 731 differentially expressed genes were screened out by mRNA-seq. GO and KEGG enrichment analysis revealed involvement in redox process and transmembrane transport. The permeability assay showed leakage of blue dye from the intestinal lumen into the body cavity. Abnormal mRNAs expression of gem-4, hmp-1, fil-2, and pho-1, which regulated intestinal development, absorption and catabolism, transmembrane transport, and apical junctions, was observed. Intestinal lipofuscin and ROS were increased, while sod-2 and isp-1 expressions were decreased. Multiple proteins in SKN-1/DAF-16 pathway were found to bind stably with gelsenicine in a predictive model. There was an up-regulation in the expression of SKN-1:GFP, while the nuclear translocation of DAF-16:GFP exhibited abnormality. The UPR biomarker HSP-4:GFP was down-regulated. In conclusion, the treatment of gelsenicine resulted in the increase of nematode intestinal permeability. The toxicological mechanisms underlying this effect involved the disruption of intestinal barrier integrity, an imbalance between oxidative and antioxidant processes mediated by the SKN-1/DAF-16 pathway, and abnormal unfolded protein reaction.
钩吻素子（Gelsemium elegans Benth.，G. elegans）是一种传统的药用植物，具有抗炎、镇痛、镇静和消肿作用。然而，它也会引起肠道副作用，如腹痛和腹泻。钩吻素子的毒理学机制尚不清楚。本研究旨在评估钩吻素子在秀丽隐杆线虫（Caenorhabditis elegans，C. elegans）中引起的肠毒性。用钩吻素子处理线虫，然后评估其生长、发育和运动行为。使用 PharmMapper 预测钩吻素子的靶标。进行 mRNA-seq 以验证预测的靶标。测量肠道通透性、ROS 生成和脂褐素积累。此外，还定量了参与氧化应激和内质网未折叠蛋白反应（UPR）的 GFP 标记蛋白的荧光强度。结果表明，钩吻素子处理导致线虫寿命抑制，以及体长、体宽和运动行为减少。通过 PharmMapper 预测到 221 个靶标，通过 mRNA-seq 筛选出 731 个差异表达基因。GO 和 KEGG 富集分析表明参与氧化还原过程和跨膜运输。通透性测定显示蓝色染料从肠腔漏入体腔。观察到调节肠道发育、吸收和分解代谢、跨膜运输和顶端连接的 gem-4、hmp-1、fil-2 和 pho-1 的异常 mRNA 表达。肠道脂褐素和 ROS 增加，而 sod-2 和 isp-1 的表达减少。在预测模型中发现 SKN-1/DAF-16 通路中的多种蛋白质与钩吻素子稳定结合。SKN-1:GFP 的表达上调，而 DAF-16:GFP 的核转位显示异常。UPR 生物标志物 HSP-4:GFP 下调。总之，钩吻素子处理导致线虫肠道通透性增加。这种作用的毒理学机制涉及肠道屏障完整性的破坏，由 SKN-1/DAF-16 通路介导的氧化和抗氧化过程之间的失衡，以及异常的未折叠蛋白反应。
