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Knockdown of CD36 inhibits leukemia cell culture supernatant-mediated platelet activation
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Abstract:
Objective: To evaluate the effect and mechanism of CD36 interference on platelet activation mediated by leukemia cell culture supernatant. Methods: Platelets were cultured with supernatant from L1210 murine leukemia cells for 4, 6, 12, and 24 hours, with platelets cultured in regular medium as the control. To determine the optimal time for supernatant-mediated platelet activation, flow cytometry was used to detect the expression of the platelet activation marker P-selectin (CD62P), and Western blot (WB) was used to detect CD36 expression. A CD36 interference vector was constructed and transfected into activated platelets. The cells were divided into the following groups: control group, model group, CD36 interference empty vector group (si-CD36 NC), CD36 interference group (si-CD36), inhibitor group (iCRT3), and inhibitor + CD36 interference group (iCRT3 + si-CD36). CCK-8 assay was used to detect platelet viability, flow cytometry was used to detect CD62P expression in platelets, and WB was used to detect the expression of PECAM-1, CD36, and β -catenin proteins in platelets. Results: The optimal time for platelet activation mediated by L1210 murine leukemia cell supernatant was 12 hours. Compared with the control group, the platelet viability, CD62P expression, and protein expression of PECAM-1, CD36, and β-catenin were significantly increased in the model group (all P < 0.01). Compared with the model group, platelet viability, CD62P expression, and protein expression of PECAM-1, CD36, and β-catenin were significantly decreased in the si-CD36 and iCRT3 groups (all P < 0.01). The changes were more pronounced in the iCRT3 + si-CD36 group compared to the iCRT3 group. Conclusion: CD36 interference inhibits β-catenin protein expression and, in combination with a Wnt/β-catenin pathway inhibitor, further inhibits murine leukemia cell supernatant-mediated platelet activation.
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