The active ingredients and mechanisms of Longchai Jiangxue Formula in treating PV, based on UPLC/Q-TOF-MS/MS, systematic pharmacology, and molecular biology validation - 17/06/21
, Weiyi Liu a , Hongwei Wu c , Yujin Li a, d , Erpeng Yang a, d , Ziqing Wang a, e , Haiyan Xiao a , Richeng Quan a, ⁎ , Xiaomei Hu a, ⁎ Bienvenido a EM-consulte, la referencia de los profesionales de la salud.
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Abstract |
Background |
Polycythemia vera (PV) is a refractory hematological disease that lack of effective therapy. Chinese traditional medicine Longchai Jiangxue formula (LCJX) has showed the powerful effects on PV. However, the active ingredients and mechanisms of this formula have not been elucidated. We explored the active ingredients and mechanisms of LCJX for treating PV.
Methods |
The chemical constituents of LCJX were qualitatively analyzed by UPLC/Q-TOF-MS/MS. On this basis, the TCMSP, ETCM, PubChem BioAssay and ChEMBL databases were searched to predict the potential targets of chemical components of LCJX. Then Genecards, GEO, DisGeNET, and OMIM databases were used to retrieve data of targets related to PV. Drug-disease-target network and protein-protein-interaction (PPI) network were built. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed. Finally, Molecular docking, CCK-8 assay, Annexin V-FITC/PI staining and western blot were processed so as to screen the active components related to PV and elucidate its mechanisms.
Results |
A total of 84 compounds were identified from LCJX by UPLC/Q-TOF-MS/MS. After removed duplicate items, there were 143 targets linked to both disease and drugs. Crucial genes, such as MTOR, HIF1A, JAK2, VEGFA, STAT3, AKT1, TERT, MAPK1, were shown in PPI network. GO enrichment indicated that oxidative stress process, tyrosine kinase activity and phosphatase binding function, and cell membrane structure were in reference to LCJX against PV. KEGG enrichment showed that JAK-STAT signaling pathway and PI3K-Akt signaling pathway, were put in an important position of the treatment. Furthermore, Molecular docking, CCK-8 assay, Annexin V-FITC/PI staining and western blot technique proved the therapeutic effect of Saikosaponin A, main ingredient of LCJX.
Conclusion |
This study, combined with UPLC/Q-TOF-MS/MS, network pharmacology and molecular biology, provides a reference for the identification of effective components, screening of quality markers and analysis of its action mechanism of LCJX.
El texto completo de este artículo está disponible en PDF.Graphical Abstract |
Highlights |
• | Traditional Chinese medicine “LongChaiJiangXue” is effective to Polycythemia Vera. |
• | The UPLC technology is useful for biochemical analysis for medicine scientifically. |
• | System pharmacology helps screen the main ingredients and mechanisms of the drugs. |
• | Hub-genes and pathways were confirmed in vitro and in silico experimentally. |
• | Molecular docking, cell viability assay and Western-blot experiment were processed. |
Abbreviations : LCJX, PV, TCMSP, PPI, GO, KEGG, STAT-3, IL-6, JAK2, VEGFA, MPN, AML, GRR, RB, AMK, IN, RPR, RRR, TCM, OB, DL, PDB, BP, MF, CC, SSA
Keywords : Longchai Jiangxue formula, UPLC/Q-TOF-MS/MS, Network pharmacology, Active ingredient, Polycythemia vera
Esquema
Vol 140
Artículo 111767- août 2021 Regresar al número
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