Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica, School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Mediators Inflamm. 2020 May 15;2020:2929163. doi: 10.1155/2020/2929163. eCollection 2020.
Relieving Sore Throat Formula (RSTF) is a formula approved by the China Food and Drug Administration and has been used for the treatment of pharyngitis in clinic for many years. However, the potential pharmacological mechanism still remains unknown. We combined multiple methods including bioinformatics data digging, network pharmacology analysis, and pathway analysis to predict the potential target of RSTF. We verified our in silico prediction results with an in vivo/vitro antibacterial effect test, mouse phagocytic index test, proliferation, transformation, and migration of mouse spleen lymphocytes. Alteration of NF-B pathway was determined by Western blotting, immunofluorescence, and PCR. The in vivo experiments demonstrated that the RSTF could significantly relieve the symptoms of pharyngitis. A rat saliva secretion test showed that RSTF can effectively relieve the xerostomia symptom. A phenol red excretion test showed that RSTF has an eliminating phlegm effect. A hot plate method and granuloma experiment proved that RSTF also have analgesic and anti-inflammatory effects. In silico prediction demonstrates that 70 active compounds of RSTF were filtered out through ADME screening and 84 putative targets correlated with different diseases. Pathway enrichment analysis showed that the candidate targets were mostly related to the response to bacteria and immunity signalling pathways, which are known contributors to pharyngitis. Experimental results confirmed that RSTF exerted therapeutic effects on pharyngitis mainly by antibacterial effect and downregulation of NF-B activities. It is demonstrated both in silico and in vivo/vitro that RSTF exerted therapeutic effects on pharyngitis mainly through an antibiotic effect and downregulation of NF-B signalling pathway.
利咽方（RSTF）是国家食品药品监督管理局批准的方剂，临床用于治疗咽炎已有多年。然而，其潜在的药理机制尚不清楚。我们结合生物信息学数据挖掘、网络药理学分析和通路分析等多种方法，预测 RSTF 的潜在靶点。我们通过体内/体外抗菌作用试验、小鼠吞噬指数试验、小鼠脾淋巴细胞增殖、转化和迁移试验对我们的计算机预测结果进行了验证。通过 Western blot、免疫荧光和 PCR 测定 NF-B 通路的变化。体内实验表明，RSTF 可显著缓解咽炎症状。大鼠唾液分泌试验表明，RSTF 能有效缓解口干症状。酚红排泄试验表明，RSTF 具有祛痰作用。热板法和肉芽肿实验证明，RSTF 还具有镇痛和抗炎作用。计算机预测表明，通过 ADME 筛选，RSTF 可筛选出 70 种活性化合物，这些化合物与 84 种与不同疾病相关的假定靶点相关。通路富集分析表明，候选靶点主要与细菌反应和免疫信号通路有关，这些通路已知与咽炎有关。实验结果证实，RSTF 对咽炎的治疗作用主要通过抗菌作用和 NF-B 活性的下调来实现。体内和体外实验均表明，RSTF 通过抗生素作用和 NF-B 信号通路的下调对咽炎发挥治疗作用。
