Thrombocytopenia is one of the most common hematological disease that can be life-threatening caused by bleeding complications. However, the treatment options for thrombocytopenia remain limited. In this study, giemsa staining, phalloidin staining, immunofluorescence and flow cytometry were used to identify the effects of 3,3ʹ-di-O-methylellagic acid 4ʹ-glucoside (DMAG), a natural ellagic acid derived from Sanguisorba officinalis L. (SOL) on megakaryocyte differentiation in HEL cells. Then, thrombocytopenia mice model was constructed by X-ray irradiation to evaluate the therapeutic action of DMAG on thrombocytopenia. Furthermore, the effects of DMAG on platelet function were evaluated by tail bleeding time, platelet aggregation and platelet adhesion assays. Next, network pharmacology approaches were carried out to identify the targets of DMAG. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to elucidate the underling mechanism of DMAG against thrombocytopenia. Finally, molecular docking simulation, molecular dynamics simulation and western blot analysis were used to explore the relationship between DAMG with its targets. DMAG significantly promoted megakaryocyte differentiation of HEL cells. DMAG administration accelerated platelet recovery and megakaryopoiesis, shortened tail bleeding time, strengthened platelet aggregation and adhesion in thrombocytopenia mice. Network pharmacology revealed that ITGA2B, ITGB3, VWF, PLEK, TLR2, BCL2, BCL2L1 and TNF were the core targets of DMAG. GO and KEGG pathway enrichment analyses suggested that the core targets of DMAG were enriched in PI3K–Akt signaling pathway, hematopoietic cell lineage, ECM-receptor interaction and platelet activation. Molecular docking simulation and molecular dynamics simulation further indicated that ITGA2B, ITGB3, PLEK and TLR2 displayed strong binding ability with DMAG. Finally, western blot analysis evidenced that DMAG up-regulated the expression of ITGA2B, ITGB3, VWF, p-Akt and PLEK. DMAG plays a critical role in promoting megakaryocytes differentiation and platelets production and might be a promising medicine for the treatment of thrombocytopenia.

中文翻译：

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DMAG，一种治疗血小板减少症的新对策

血小板减少症是最常见的血液系统疾病之一，可因出血并发症而危及生命。然而，血小板减少症的治疗选择仍然有限。在这项研究中，使用吉姆萨染色、鬼笔环肽染色、免疫荧光和流式细胞术来确定 3,3′-二-O-甲基鞣花酸 4′-葡萄糖苷 (DMAG) 的作用，DMAG 是一种源自地榆的天然鞣花酸。 (SOL ) 对 HEL 细胞中的巨核细胞分化。然后，通过X射线照射构建血小板减少症小鼠模型，以评估DMAG对血小板减少症的治疗作用。此外，通过尾部出血时间、血小板聚集和血小板粘附试验评估 DMAG 对血小板功能的影响。接下来，进行网络药理学方法来识别 DMAG 的目标。进行基因本体论 (GO) 和京都基因和基因组百科全书 (KEGG) 通路富集分析，以阐明 DMAG 对抗血小板减少症的基本机制。最后，利用分子对接模拟、分子动力学模拟和蛋白质印迹分析来探索DAMG与其靶点之间的关系。DMAG显着促进HEL细胞的巨核细胞分化。DMAG 给药加速血小板恢复和巨核细胞生成，缩短尾出血时间，增强血小板聚集和血小板减少小鼠的粘附。网络药理学显示，ITGA2B、ITGB3、VWF、PLEK、TLR2、BCL2、BCL2L1和TNF是DMAG的核心靶点。GO和KEGG通路富集分析表明DMAG的核心靶点富含PI3K-Akt信号通路，造血细胞谱系、ECM-受体相互作用和血小板活化。分子对接模拟和分子动力学模拟进一步表明，ITGA2B、ITGB3、PLEK和TLR2与DMAG具有很强的结合能力。最后，蛋白质印迹分析证明 DMAG 上调了 ITGA2B、ITGB3、VWF、p-Akt 和 PLEK 的表达。DMAG 在促进巨核细胞分化和血小板生成中起关键作用，可能是治疗血小板减少症的有前途的药物。