[关键词]
[摘要]
目的 基于网络毒理学策略,研究商陆致大鼠肾损伤的潜在作用机制。方法 通过查阅在线数据库并挖掘文本,建立商陆的化学成分库;基于药效团的反向分子对接技术,预测化学成分的相关靶标,并与肾毒性靶标交集投射,得到商陆致大鼠肾毒性的作用靶标,并反推获得商陆致肾毒性的潜在物质基础;利用大规模蛋白互作筛选关键靶标,并通过GO和KEGG生物学注释分析肾毒性关键通路。构建商陆皂苷甲诱导肾毒性的大鼠模型,并采用分子生物学方法检测相关通路的重要靶标表达情况。结果 通过文献和相关数据库共得到56个化学成分和148个潜在靶标,其中38个成分,34个靶标和93条通路与肾毒性的产生极为密切,主要涉及TNF信号通路、钙离子信号通路、NF-κB信号通路、VEGF信号通路等相关分子环节;病理结果显示,给予商陆皂苷甲7 d后,大鼠肾脏组织发生了不同程度的损伤;Western blotting法检测显示核转录因子-κB(NF-κB)抑制蛋白α(IκBα)蛋白表达下调(P<0.01),p-IκBα蛋白表达上调(P<0.05);ELISA法定量检测结果显示商陆皂苷甲使大鼠血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)水平显著升高(P<0.05)。结论 网络毒理学可用于初步筛选潜在毒性物质基础,基于网络毒理学筛选的商陆皂苷甲可诱导大鼠肾毒性,其分子机制与激动NF-κB信号通路,过表达炎性因子有关。
[Key word]
[Abstract]
Objective Based on the strategy of network toxicology, the potential mechanism of renal injury in rats induced by Phytolaccae Radix was preliminarily explored. Methods Firstly, by consulting the online database and mining the text, the chemical composition library of Phytolaccae Radix was established. The reverse molecular docking technology based on pharmacophores was performed to predict the relevant targets of chemical components and the target of nephrotoxicity induced by Phytolaccae Radix was obtained by intersecting with the target of nephrotoxicity. Potential substance basis of nephrotoxicity induced by Phytolaccae Radix was obtained by reverse analysis. Secondly, a large-scale protein-protein interactions were used to screen key targets, and key pathways for nephrotoxicity were analyzed by GO and KEGG biological annotation. Finally, a rat model of nephrotoxicity induced by esculentoside A was constructed and molecular biology methods were used to detect the expression of important targets in related pathways. Results A total of 56 chemical components and 148 potential targets were obtained through literatures and related databases. Among them, 38 components, 34 targets, and 93 pathways were closely related to the generation of nephrotoxicity, mainly involving TNF signaling pathway, calcium signaling pathway, NF-κB signaling pathway, and VEGF signaling, thus participating in the beginning and end of nephrotoxicity events. Pathological results showed that the kidney tissues of rats were damaged to varying degrees after 7 d of treatment with esculentoside A. Western blot showed that the expression of IκBα was down-regulated (P < 0.01) and the expression of p-IκBα was up-regulated (P < 0.05). Quantitative detection of TNF-α and IL-1β by ELISA showed that both expressions were up-regulated (P < 0.05). Conclusion Network toxicology can be used to preliminarily screen potential toxic substances. Esculentoside A based on network toxicology screening can induce nephrotoxicity in rats. Its molecular mechanism is related to activation of NF-κB signaling pathway and overexpression of inflammatory factors.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(81173541)