目的：评估干扰CD36表达白血病细胞培养上清液对血小板活化的影响及其机制。方法: 利用L1210小鼠白血病细 胞上清液培养血小板 4、6、12、24 h，以普通培养基培养血小板作为对照，通过流式细胞术检测血小板活化标志物 P-选择素 （CD62P）的表达，WB法检测CD36表达，确定上清液活化血小板最佳时间。构建CD36干扰载体转染至活化的血小板中，实验分 为对照组、模型组、CD36干扰空载体组（si-CD36 NC）、CD36干扰组（si-CD36）、抑制剂组（iCRT3）、抑制剂 + CD36干扰组（iCRT3 + si-CD36），CCK-8法检测血小板活力，流式细胞术检测血小板中CD62P表达，WB法检测血小板中PECAM-1、CD36、β-catenin蛋 白表达。结果: L1210小鼠白血病细胞上清液活化血小板最佳时间为12 h。与对照组相比，模型组血小板活力、CD62P表达、 PECAM-1、CD36、β-catenin 蛋白表达均显著上升（均 P < 0.01）。与模型组相比，si-CD36 和 iCR73 组血小板活力、CD62P 表达、 PECAM-1、CD36、β-catenin 蛋白表达均显著下降（均 P < 0.01）。与 iCRT3 组相比，iCRT3 + si-CD36 组变化更为显著。结论: CD36干扰抑制β-catenin蛋白表达，协同Wnt/β-catenin通路抑制剂，进而抑制小鼠白血病细胞上清液介导的血小板活化。

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.

武汉市卫健委基金项目（No.WG20D09）