槲皮素-3-葡萄糖醛酸苷通过PI3K/AKT通路对免疫性骨髓衰竭小鼠血小板凋亡和活化的影响  

Effects of Quercetin-3-O-β-D-Glucuronide on Platelet Apoptosis and Activation in Mice with Immune-Mediated Bone Marrow Failure via PI3K/AKT Pathway

作　　者：夏乐敏 余海 金喆 丁洁 余和平 XIA Le-Min;YU Hai;JIN Zhe;DING Jie;YU He-Ping(Department of Hematology,Jing’an District Centre Hospital of Shanghai(Jing’an Branch,Huashan Hospital,Fudan University),Shanghai 200040,China)

机构地区：[1]上海市静安区中心医院(复旦大学附属华山医院静安分院)血液科,上海200040

出　　处：《中国实验血液学杂志》2023年第5期1437-1442,共6页Journal of Experimental Hematology

基　　金：上海市静安区卫生健康系统医学科研课题(2019MS02);上海市卫生健康委员会卫生行业临床研究专项项目(202140034);上海市西部区域中医联合体及上海市中医医院分院2022年度“未来计划”中医药传承项目(XBYLT-WLJH-2023-006)。

摘　　要：目的:探讨槲皮素-3-葡萄糖醛酸苷(QG)通过PI3K/AKT通路对免疫性骨髓衰竭小鼠模型血小板凋亡和活化的影响。方法:建立免疫性骨髓衰竭小鼠模型,将40只C57BL/6小鼠随机分为正常、模型、环孢素(CsA)、QG共4组,每组10只。正常组及模型组小鼠分别给予生理盐水灌胃,CsA组给予0.027 g/kg CsA灌胃,QG组给予0.2 g/kg QG灌胃。造模3 d后处死小鼠并从尾静脉处抽血,用全自动血细胞分析仪测定血小板数并制备洗涤血小板。采用流式细胞仪检测BAX、BAD、caspase-9、磷脂酰丝氨酸(PS)、血小板激活复合物-1(PAC-1)、P-选择素的表达;使用Western blot法测定PI3K、p-PI3K、AKT、p-AKT的蛋白含量;运用实时定量PCR(RT-qPCR)法检测PI3K mRNA、AKT mRNA的表达。结果:与正常组相比较,模型组中小鼠外周血小板数明显减少,洗涤血小板中的BAX、BAD、caspase-9、PS、PAC-1、P-选择素明显升高(P<0.05),同时,PI3K、p-PI3K、AKT和p-AKT蛋白含量以及PI3K mRNA和AKT mRNA表达水平则明显下降(P<0.05);与模型组相比较,CsA组及QG组洗涤血小板中的BAX、BAD、caspase-9、PS、PAC-1、P-选择素明显下降(P<0.05),而PI3K、p-PI3K、AKT和p-AKT蛋白水平以及PI3K mRNA及AKT mRNA表达水平则明显上升(P<0.05)。结论:QG能减少免疫性骨髓衰竭小鼠血小板凋亡,并参与部分血小板活化功能的调节,其机制可能与PI3K/AKT通路有关。Objective:To investigate the effect of quercetin-3-O-β-D-glucuronide(QG)on platelet apoptosis and activation in mice with immune-mediated bone marrow failure via PI3K/AKT pathway.Methods:An immune-mediated bone marrow failure mice model was established and forty C57BL/6 mice were randomly assigned into 4 groups:normal group,model group,cyclosporine(CsA)group and QG group,with 10 mice in each group.The mice in CsA group were intragastrically administered with 0.027 g/kg CsA daily and the mice in QG group were intragastrically administered with 0.2 g/kg QG daily,while the mice in the normal group and model group were intragastrically administered with normal saline,respectively.After three days of modeling,the mice were euthanized,and the blood was collected through the tail vein.Part of the blood was used for blood routine examination,and the other part was used to prepare washed platelets.Some of the prepared washed platelets were used to detect the expressions of BAX,BAD,caspase-9,phosphatidylserine(PS),platelet activated complex-1(PAC-1)and P-selectin by flow cytometry;some of washed platelets was used to determine the protein contents of PI3K,p-PI3k,AKT and p-AKT by Western blot;the other part of the washed platelets was used to measure the expressions of PI3K mRNA and AKT mRNA by real-time quantitative PCR(RT-qPCR).Results:In the model group,the absolute platelet count of the mice was significantly decreased,and the levels of BAX,BAD,caspase-9,PS,PAC-1,and P-selectin in the washed platelets were significantly increased,compared to the normal group(P<0.05).At the same time,the expression levels of PI3K,p-PI3K,AKT,p-AKT proteins and PI3K mRNA,AKT mRNA in model group were significantly reduced,compared to the normal group(P<0.05).In the CsA group and QG group,the expression levels of BAX,BAD,caspase-9,PS,PAC-1,and P-selectin in the washed platelets were significantly reduced(P<0.05),while the levels of PI3K,p-PI3K,AKT,p-AKT and PI3K mRNA,AKT mRNA were significantly increased,compared to the model group(P<0.05).

关 键 词：槲皮素-3-葡萄糖醛酸苷 PI3K/AKT 血小板 凋亡 活化 

分 类 号：R551.3[医药卫生—血液循环系统疾病]