Thrombopoietin and collagen in low doses cooperatively induce human platelet activation

• Published in: Acute medicine & surgery 2022-01, Vol.9 (1), p.e769-n/a
• Main Authors: Doi, Tomoaki, Hori, Takamitsu, Onuma, Takashi, Mizutani, Daisuke, Ueda, Kyohei, Enomoto, Yukiko, Matsushima‐Nishiwaki, Rie, Tanabe, Kumiko, Hioki, Tomoyuki, Tokuda, Haruhiko, Iwama, Toru, Iida, Hiroki, Kozawa, Osamu, Ogura, Shinji
• Format: Article
• Language: English
• Subjects: Aggregates; Antibodies; Coagulopathy; Collagen; Drug dosages; Growth factors; human platelets; Original; Plasma; Proteins; Sepsis; Thrombocytopenia; thrombopoetin
• ISSN: 2052-8817; 2052-8817
• DOI: 10.1002/ams2.769

Aim In acute medicine, we occasionally treat life‐threatening conditions such as sepsis and trauma, which cause severe thrombocytopenia. Serum thrombopoietin levels have been reported to increase under the condition of thrombocytopenia related to severity. Collagen is a crucial activator of platelets, and Rho family members, such as Rho/Rho‐kinase and Rac, play roles as active molecules involved in the intracellular signaling pathways in platelet activation. The present study aimed to elucidate the effects of thrombopoietin (TPO) on subthreshold low‐dose collagen‐stimulated human platelets in terms of Rho/Rho‐kinase and Rac. Methods Platelet‐rich plasma donated from healthy volunteers was stimulated by the subthreshold low‐dose of collagen after pretreatment with TPO and/or NSC23766, an inhibitor of the Rac‐guanine nucleotide exchange factor interaction, or Y27632, an inhibitor of Rho‐kinase. Platelet aggregation was measured using an aggregometer based on laser‐scattering methods. Proteins involved in intracellular signaling were analyzed using western blotting, and the secretion of platelet‐derived growth factor‐AB from activated platelets was determined using an enzyme‐linked immunosorbent assay. Results Under the existence of TPO, the low dose of collagen remarkably elicited the aggregation and platelet‐derived growth factor‐AB secretion of platelets, which were suppressed by NSC23766 and Y27632. The combination of TPO and collagen considerably induced a transient increase of guanosine triphosphate (GTP)‐binding Rac and GTP‐binding Rho followed by an increase of phosphorylated cofilin, a Rho‐kinase substrate. Conclusion These results strongly suggest that TPO and collagen in low doses cooperatively potentiate human platelet activation through both Rac and Rho/Rho‐kinase mediated pathways. Thrombopoietin and collagen in low doses cooperatively potentiate human platelet activation via both Rac and Rho/Rho‐kinase mediated pathways. The effect of TPO on platelet activation under the stimulation of collagen, even though of which the levels are insufficient to trigger by itself, might indicate a compensatory mechanism to enhance platelet activation in thrombocytopenia.