基于“病-方”互作网络探索尪痹片干预膝骨关节炎临床优势分期与潜在作用机制

Clinical advantage staging and underlying mechanisms of Wangbi Tablets against knee osteoarthritis based on"disease-formula"interaction network

从“病-方”互作网络探索尪痹片干预膝骨关节炎(knee osteoarthritis,KOA)临床优势分期及潜在作用机制。首先,基于临床指南和专家共识并结合中医证候本体及多维定量关联计算平台(SoFDA)分别收集KOA发作期、缓解期和康复期3个临床分期及尪痹片相关症状及基因集,同时基于中医药百科全书数据库(ETCM 2.0)收集尪痹片所含成分候选靶标谱,分别获得疾病靶标分子库和方药候选靶标集。其次,采用Jaccard和余弦相似度分别对“药物治疗与疾病临床症状”“药靶与疾病基因”“富集通路”间相似度评价,在尪痹片干预KOA“病-方”互作关联网络构建与分析后,提取方药干预不同临床分期网络,挖掘其作用特点。最后,采用脂多糖(LPS)诱导SW1353细胞考察方药作用机制。结果显示,基于SoFDA收集到KOA发作期、缓解期和康复期及尪痹片临床治疗症状及基因92/3921、138/3708、139/3800、196/3946个,基于ETCM 2.0得到260个尪痹片所含化学成分预测靶标集。“疾病分期-药物”对应性评价发现,尪痹片与KOA缓解期在“药物治疗与疾病临床症状”“药靶与疾病基因”“富集通路”3个层次具有最高相似度,其次为康复期;网络构建分析发现,尪痹片的核心效应靶标共同矫正不同分期机体“免疫-炎症”失衡,并发挥镇痛和骨保护作用,缓解膝关节疼痛、关节肿胀、酸痛、乏力、功能障碍等症状。在KOA发作期和缓解期方药核心效应靶标尤其发挥抗氧化作用,在康复期方药核心效应靶标发挥调节物质与能量代谢紊乱及血管保护作用。基于体外细胞模型发现,尪痹片通过磷脂酰肌醇-3-激酶(PI3K)-蛋白激酶B(Akt)信号通路可抑制白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)等炎症因子释放及Bax/Bcl-2表达。该研究为进一步明确尪痹片临床优势分期与辅助临床精准定位提供了理论依据。

The clinical advantage staging and underlying mechanisms of Wangbi Tablets against knee osteoarthritis(KOA) were studied based on the "disease-formula" interaction network. Firstly, the clinical symptoms and related genes corresponding to Wangbi Tablets and KOA in the acute, remission, and recovery phases were collected from clinical guidelines/consensus and SoFDA database, and the putative targets of Wangbi Tablets were obtained from ETCM 2.0. Then, Jaccard similarity and cosine similarity were employed to assess the similarities of clinical symptoms, genes, and enriched pathways between Wangbi Tablets and KOA in different phases. The "disease-formula" interaction network of the drug targets and disease genes was constructed, and the key targets were screened by topological feature calculation. KEGG and Reactome database were used for the functional enrichment of the key targets, on the basis of which the functional characteristics of Wangbi Tablets against KOA in the acute, remission, and recovery phases were predicted. Finally, the SW1353 cells exposed to lipopolysaccharide were used to decipher the mechanism of Wangbi Tablets against KOA. The results showed that 92/3 921, 138/3 708, 139/3 800, and 196/3 946 clinical symptoms and the related genes corresponded to KOA in the acute, remission, and recovery phases and Wangbi Tablets were collected from SoFDA, and 260 putative targets of Wangbi Tablets were obtained from ETCM 2.0. Wangbi Tablets had highest similarity of clinical symptoms, genes, and enriched pathways with KOA in the remission phase and the secondary highest similarity with KOA in the recovery phase. The key targets of Wangbi Tablets mainly participated in the regulation of immunity-inflammation imbalance and exerted pain-relieving and bone-protecting effects to alleviate symptoms such as knee joint pain, joint swelling, soreness, fatigue, and dysfunction. Intriguingly, the key targets of Wangbi Tablets possessed antioxidant effects during KOA in the acute and remission phases, while they maintained material and energy metabolism homeostasis and protected vessels during KOA in the recovery phase. The cell experiment indicated that Wangbi Tablets down-regulated the expression of interleukin(IL)-6, IL-1β, tumor necrosis factor-α(TNF-α), and Bcl-2-associated X protein(Bax)/B-cell lymphoma 2(Bcl-2) via regulating the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt) signaling pathway. The findings lay a theoretical foundation for further clarifying the clinical advantage stage and precise clinical application of Wangbi Tablets in treating KOA.