Network pharmacology, molecular docking and experimental verification of the mechanism of huangqi-jixuecao herb pair in treatment of peritoneal fibrosis
Network pharmacology, molecular docking and experimental verification of the mechanism of huangqi-jixuecao herb pair in treatment of peritoneal fibrosis
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DOI：
10.1016/j.jep.2023.116874
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发表时间：
2023-07-20
影响因子：
5.4
通讯作者：
Sheng，Meixiao
中科院分区：
文献类型：
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作者：
Dai，Huibo;Shan，Yun;Sheng，Meixiao
民族药理学相关性黄芪-积血草对（HQJXCHP）是由黄芪（黄芪）和积雪草（积雪草）两种广泛应用的中药方剂组成的传统草药配方，数百年来一直用于补气清热。然而，目前HQJXCHP抗腹膜纤维化（PF）的治疗效果和潜在靶点尚不清楚。研究目的本研究的主要目的是基于网络药理学分析和实验验证，初步预测和验证HQJXCHP抗腹膜纤维化（PF）的作用和分子机制。材料与方法通过HPLC-Q-TOF/MS对HQJXCHP的成分进行分析。用于网络药理学分析的HQJXCHP的生物活性化合物是从TCMSP数据库中获得的。 PF 中 HQJXCHP 相关的治疗靶点是从 GeneCards、OMIM、Therapeutic Targets 和 PharmGkb 数据库中获得的。通过 GO 和 KEGG 通路富集分析预测治疗靶点相关信号通路。在PDS诱导的PF小鼠体内模型和PMCs MMT体外模型中进一步验证了HQJXCHO的靶点。结果共检索到HQJXCHP相关188个靶基因的23个生物活性化合物。 HQJXCHP 复合靶点和 PF 相关靶点网络识别出 131 个常见靶点基因。随后的蛋白质-蛋白质相互作用（PPI）网络分析结果显示Akt1、TP53、TNF、VEGFA和CASP3为HQJXCHP的前五个关键靶点。进一步的分子对接数据显示，HQJXCHP 的两种关键化合物槲皮素和山奈酚对这些关键靶点具有很强的亲和力。 GO和KEGG通路富集分析进一步表明PI3K/Akt、IL-17、TNF和TLR通路有助于HQJXCHP对PF的治疗作用。采用体内PDS诱导的PF小鼠模型和体外PMCs间皮间质转化（MMT）模型（有或没有HQJXCHP干预）来证实HQJXCHP的作用和作用机制。 Western blot和qRT-PCR结果显示，HQ、JXC和HQJXCHP通过抑制MMT过程减少PDS诱导的炎症细胞聚集和腹膜增厚，其中HQJXCHP发挥最大的治疗效果。此外，HQJXCHP还能抑制PDS诱导的PI3K/Akt、IL-17、TNF和TLR信号通路的激活。结论本研究首次采用网络药理学和分子对接分析来预测HQJXCHP对PDS相关PF治疗作用的靶点。体内和体外验证实验的数据共同表明，HQJXCHP通过抑制PI3K/Akt、IL-17、TNF和TLR信号通路来延迟PF过程。总的来说，我们的研究结果强调了网络药理学理论的成功应用，为 HQJXCHP 治疗 PF 的临床应用提供了科学依据。
Ethnopharmacological relevanceThe Huangqi-Jixuecao herb pair (HQJXCHP) is a traditional herbal formula composed of two widely applied TCM prescriptions, Huangqi (Astragalus membranaceus (Fisch.) Bunge) and Jixuecao (Centella asiatica (L.) Urb.), used for hundreds of years to replenish qi and clear away heat. However, the therapeutic effects of HQJXCHP against peritoneal fibrosis (PF) and potential targets are currently unclear.Aims of the studyThe main objective of this study was preliminary prediction and validation of the effects and molecular mechanisms of action of HQJXCHP against PF based on network pharmacology analysis and experimental verification.Materials and methodsThe ingredients of HQJXCHP were analyzed via HPLC-Q-TOF/MS. Bioactive compounds of HQJXCHP used for network pharmacology analysis were obtained from the TCMSP database. HQJXCHP-related therapeutic targets in PF were obtained from the GeneCards, OMIM, Therapeutic Targets and PharmGkb databases. Therapeutic target-related signaling pathways were predicted via GO and KEGG pathway enrichment analyses. The targets of HQJXCHO were further validated in a PDS-induced PF mouse modelin vivoand PMCs MMT modelin vitro.ResultsA total of 23 bioactive compounds of HQJXCHP related 188 target genes were retrieved. The HQJXCHP compound-target and PF-related target networks identified 131 common target genes. Subsequent protein-protein interaction (PPI) network analysis results disclosed Akt1, TP53, TNF, VEGFA and CASP3 as the top five key targets of HQJXCHP. Further molecular docking data revealed strong affinity of the two key compounds of HQJXCHP, quercetin and kaempferol, for these key targets. GO and KEGG pathway enrichment analyses further showed that PI3K/Akt, IL-17, TNF and TLR pathways contribute to the therapeutic effects of HQJXCHP on PF. Anin vivoPDS-induced PF mouse model andin vitroPMCs mesothelial-to-mesenchymal transition (MMT) model with or without HQJXCHP intervention were used to confirm the effects and mechanisms of action of HQJXCHP. Western blot and qRT-PCR results showed that HQ, JXC and HQJXCHP reduced PDS-induced inflammatory cell aggregation and peritoneal thickening through suppressing the MMT process, among which HQJXCHP exerted the greatest therapeutic effect. Moreover, HQJXCHP inhibited activation of the PI3K/Akt, IL-17, TNF and TLR signaling pathways induced by PDS.ConclusionsThis is the first study to employ network pharmacology and molecular docking analyses to predict the targets of HQJXCHP with therapeutic effects on PDS-related PF. Data fromin vivoandin vitrovalidation experiments collectively showed that HQJXCHP delays the PF process through inhibiting PI3K/Akt, IL-17, TNF and TLR signaling pathways. Overall, our findings highlight the successful application of network pharmacology theory to provide a scientific basis for clinical utility of HQJXCHP against PF.