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16 reviewsabstractArticle history:Radiation-induced thrombocytopenia (RIT) faces a perplexing challenge in the clinical treatment of cancerReceived 3 February 2024patients, and current therapeutic approaches are inadequate in the clinical settings. In this research, oxyReceived in revised formmatrine, a new molecule capable of healing RIT was screened out, and the underlying regulatory mecha17 July 2024nism associated with magakaryocyte (MK) differentiation and thrombopoiesis was demonstrated. TheAccepted 22 July 2024capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro. TheAvailable online 23 July 2024ability to induce thrombopoiesis was subsequently demonstrated in Tg (cd41:enhanced green fluorescentKeywords:protein (eGFP)) zebrafish and RIT model mice. In addition, we carried out network pharmacological prediction, drug affinity responsive target stability assay (DARTS) and cellular thermal shift assay (CETSA)Oxymatrineanalyses to explore the potential targets of oxymatrine. Moreover, the pathway underlying the effects ofMegakaryocyte differentiationoxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses,ThrombopoiesisWestern blot (WB), and immunofluorescence. Oxymatrine markedly promoted MK differentiation andPlateletsToll-like receptor 2maturation in vitro. Moreover, oxymatrine induced thrombopoiesis in Tg (cd41:eGFP) zebrafish andaccelerated thrombopoiesis and platelet function recovery in RIT model mice. Mechanistically, oxymatrinedirectly binds to toll-like receptor 2 (TLR2) and further regulates the downstream pathway stimulator ofinterferon genes (STING)/nuclear factor-kappaB (NF-kB), which can be blocked by C29 and C-176, which arespecific inhibitors of TLR2 and STING, respectively. Taken together, we demonstrated that oxymatrine, anovel TLR2 agonist, plays a critical role in acceleratingMK
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