Network Pharmacology-Based Mechanism Prediction and Pharmacological Validation of Xiaoyan Lidan Formula on Attenuating Alpha-Naphthylisothiocyanate Induced Cholestatic Hepatic Injury in Rats.
Network Pharmacology-Based Mechanism Prediction and Pharmacological Validation of Xiaoyan Lidan Formula on Attenuating Alpha-Naphthylisothiocyanate Induced Cholestatic Hepatic Injury in Rats.
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DOI：
10.1016/j.jep.2021.113816
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发表时间：
2021-01
影响因子：
5.4
通讯作者：
Meiqi Wang;Fangle Liu;Yufeng Yao;Qiuyu Zhang;Zenghui Lu;Runjing Zhang;Changhui Liu;Chaozhan Lin;Chenchen Zhu
中科院分区：
文献类型：
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作者：
Meiqi Wang;Fangle Liu;Yufeng Yao;Qiuyu Zhang;Zenghui Lu;Runjing Zhang;Changhui Liu;Chaozhan Lin;Chenchen Zhu
中药小炎利胆方（XYLDF）具有清热、祛湿、祛黄的功效。长期以来，它在临床上被用于治疗肝内胆汁淤积（IHC）引起的肝胆疾病。然而，XYLDF的治疗作用机制尚不清楚。本研究旨在通过网络药理学和实验分析，探讨XYLDF对anit诱导的胆汁淤积性肝损伤（CHI）大鼠的潜在肝脏保护作用机制。材料与方法在UPLC-TOF-MS/MS鉴定的29个XYLDF血清迁移化合物的基础上，应用网络药理学方法对其作用机制进行预测。系统的网络被构建来识别潜在的分子靶点、生物过程和信号通路。通过分子对接模拟具有显著潜力的靶点与活性化合物之间的相互作用。为验证其作用机制，采用anit诱导大鼠模型，通过血清生化、胆汁流速、组织病理学检查，以及RT-qPCR和WB分析肝脏和回肠胆汁酸合成、输出、输入相关酶等基因和蛋白的表达，以及炎症因子的表达，评价XYLDF对CHI的影响。结果网络药理学研究结果表明，TNF （TNF-α）、RELA （NF-κB）、NR1H4 （FXR）、ICAM1 （ICAM-1）是XYLDF治疗胆汁淤积性肝损伤的重要潜在靶点，与胆汁代谢和NF-κB介导的炎症信号转导有关。并且分子对接预先验证了网络药理学的预测，因为XYLDF的核心活性化合物与FXR具有较强的模拟结合亲和力，其次是NF-κB、TNF-α和ICAM-1。同时，口服XYLDF对anit诱导的CHI大鼠的作用表现为降低血清转氨酶（ALT、AST）、TBA和TBIL水平，提高胆汁流速，显著改善肝脏组织病理学。此外，与上述靶标预测和分子对接一致，XYLDF显著上调肝脏中FXR、SHP、BSEP和MRP2的表达，下调CYP7A1和NTCP的表达，促进回肠中IBABP和OSTα/β的表达，提示激活了FXR介导的胆汁酸合成、转运和重吸收途径。此外，XYLDF治疗后，血浆和肝脏中TNF-α水平降低，肝脏中NF-κB、TNF-α、ICAM-1基因和蛋白表达降低，表明NF-κB介导的炎症信号通路受到抑制，这可以通过抑制NF-κB的核易位来证明。结论syldf对抗抗肽诱导的胆汁淤积性肝损伤具有改善的肝保护作用。本研究证实其机制为激活fxr调控的胆汁酸通路，通过NF-κB信号通路抑制炎症。
Ethnopharmacological relevanceThe well-known Chinese prescription, Xiaoyan Lidan Formula (XYLDF), possesses efficiency of heat-clearing, dampness-eliminating and jaundice-removing. It has long been used clinically for the treatment of hepatobiliary diseases due to intrahepatic cholestasis (IHC). However, the mechanism of XYLDF for its therapeutic effects remains elusive.Aim of the studyThe study aimed to explore the potential targets for liver protective mechanism of XYLDF based on network pharmacology and experimental assays in ANIT-induced cholestatic hepatic injury (CHI) in rats.Materials and methodsOn the basis of the 29 serum migrant compounds of XYLDF elucidated by UPLC-TOF-MS/MS, a network pharmacology approach was applied for the mechanism prediction. Systematic networks were constructed to identify potential molecular targets, biological processes, and signaling pathways. And the interactions between significantly potential targets and active compounds were simulated by molecular docking. For the mechanism validation, an ANIT-induced rat model was used to evaluate the effects of XYLDF on CHI according to serum biochemistry, bile flow rates, histopathological examination, and the gene and protein expression including enzymes related to synthesis, export, and import of bile acid in liver and ileum, and those of inflammatory cytokines, analyzed by RT-qPCR and WB.ResultsThe results of network pharmacology research indicated TNF (TNF-α), RELA (NF-κB), NR1H4 (FXR), and ICAM1 (ICAM-1) to be the important potential targets of XYLDF for cholestatic liver injury, which are related to bile metabolism and NF-κB-mediated inflammatory signaling. And the molecular docking had pre-validated the prediction of network pharmacology, as the core active compounds of XYLDF had shown strong simulation binding affinity with FXR, followed by NF-κB, TNF-α, and ICAM-1. Meanwhile, the effects of XYLDF after oral administration on ANIT-induced CHI in rats exhibited the decreased levels of transaminases (ALT and AST), TBA, and TBIL in serum, raised bile flow rates, and markedly improved hepatic histopathology. Furthermore, consistent to the above targets prediction and molecular docking, XYLDF significantly up-regulated the expression of FXR, SHP, BSEP, and MRP2, and down-regulated CYP7A1 and NTCP in liver, and promoted expression of IBABP and OSTα/β in ileum, suggesting the activation of FXR-mediated pathway referring to bile acid synthesis, transportation, and reabsorption. Moreover, the lower levels of TNF-α in plasma and liver, as well as the reduced hepatic gene and protein expression of NF-κB, TNF-α, and ICAM-1 after XYLDF treatment revealed the suppression of NF-κB-mediated inflammatory signaling pathway, as evidenced by the inhibition of nuclear translocation of NF-κB.ConclusionsXYLDF exhibited an ameliorative liver protective effect on ANIT-induced cholestatic hepatic injury. The present study has confirmed its mechanism as activating the FXR-regulated bile acid pathway and inhibiting inflammation via the NF-κB signaling pathway.