杨梅苷对辐射所致血小板减少症的治疗作用及分子机制  

Therapeutic Effect and Molecular Mechanism of Myricetin on Radiation-Induced Thrombocytopenia

作　　者：邓君竹 吴建明 易泰安 李跃跃 胡海洋 邹文俊[1] DENG Junzhu;WU Jianming;YI Taian;LI Yueyue;HU Haiyang;ZOU Wenjun(Chengdu University of Traditional Chinese Medicine,Chengdu 610075;Southwest Medical University,Luzhou 646000)

机构地区：[1]成都中医药大学,成都610075 [2]西南医科大学,泸州646000

出　　处：《中药药理与临床》2023年第5期56-62,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:82074129);四川省杰出青年基金项目(编号:2022JDJQ0061)。

摘　　要：目的:研究杨梅苷对K562细胞向巨核细胞分化和X射线致血小板减少症小鼠的治疗作用及分子机制。方法:杨梅苷干预K562细胞后,利用镜下拍照观察细胞形态变化;吉姆萨染色观察多倍体形成情况;鬼笔环肽染色观察纤维型肌动蛋白的表达;流式细胞术检测巨核细胞表面抗原CD41^(+)/CD42b^(+)的表达,从而验证杨梅苷促进巨核细胞分化的活性。采用4Gy X射线辐射昆明小鼠建立血小板减少症模型,将造模后的小鼠随机分为模型对照组、杨梅苷5、10 mg/kg组、阳性药rhTPO 3 000 U/kg组,给药第4、7、11 d时使用血细胞分析仪检测小鼠外周血象。给药第11 d, HE染色观察小鼠骨髓巨核细胞数目;流式细胞术检测脾脏细胞CD41^(+)/CD61^(+)表达水平。杨梅苷干预K562细胞后,蛋白印迹法检测MEK和ERK蛋白及磷酸化蛋白的表达。结果:与空白对照组相比,杨梅苷10、20、40μmol/L组可显著促进K562细胞体积增大,多倍体细胞数目增多,纤维型肌动蛋白荧光强度增强,CD41^(+)/CD42b^(+)表达升高(P<0.01);与模型对照组相比,杨梅苷5、10 mg/kg组可显著加速X射线诱导的血小板减少症小鼠血小板浓度的恢复(P<0.05或P<0.01),增强骨髓有核细胞数以及脾细胞CD41^(+)/CD61^(+)的表达(P<0.01);杨梅苷10、20、40μmol/L组明显上调K562细胞MEK和ERK的磷酸化蛋白表达(P<0.05或P<0.01)。结论:杨梅苷可通过激活MEK/ERK信号通路促进巨核细胞分化,促进X射线诱导的血小板减少症小鼠的血小板生成。Objective:To study the therapeutic effect and molecular mechanism of myricetin on megakaryocyte differentiation of K562 cells and thrombocytopenia induced by X-ray radiation in mice.Methods:After intervention with myricetin on K562 cells,cell morphology changes were observed under a microscope,and polyploid formation was examined by Giemsa staining.F-actin expression was observed by phalloidin staining,and the expression of megakaryocyte surface antigens CD41^(+)/CD42b^(+)was detected by flow cytometry to validate the activity of myricetin in promoting megakaryocyte differentiation.A thrombocytopenia model was established in Kunming mice by irradiation with 4 Gy Xrays.After modeling,mice were randomly divided into a model group,high-and low-dose myricetin groups(10 and 5 mg/kg),and a positive drug group(rhTPO,3000 U/kg).On the 4th,7th,and 11th day of drug administration,the peripheral blood cells of mice were analyzed using a blood cell analyzer.On the 11th day of drug administration,the number of megakaryocytes in mice bone marrow was observed by H&E staining,and the expression level of CD41^(+)/CD61^(+)in the spleen was detected by flow cytometry.After intervention with myricetin on K562 cells,the expression levels of MEK and ERK proteins were detected by Western blot.Results:Compared with the blank control group,myricetin significantly promoted the enlargement of K562 cell volume,increased the number of polyploid cells,enhanced F-actin fluorescence intensity,and up-regulated the expression of CD41^(+)/CD42b^(+)(P<0.01).Compared with the model group,myricetin significantly accelerated the recovery of platelet levels in thrombocytopenic mice induced by X-ray radiation(P<0.05 or P<0.01),the number of bone marrow nucleated cells,and the expression of CD41^(+)/CD61^(+)in spleen cells(P<0.01).Moreover,myricetin significantly up-regulated the phosphorylation levels of MEK and ERK in K562 cells(P<0.05 or P<0.01).Conclusion:Myricetin can promote megakaryocyte differentiation by activating the MEK/ERK signaling pathway

关 键 词：杨梅苷 血小板减少症 巨核细胞分化 细胞外调节激酶 分裂原活化蛋白激酶 

分 类 号：R285[医药卫生—中药学]