【作者】 张鹏； 乔昆； 任雨笙； 梁春； 冷冰； 吴宗贵；

【Author】 ZHANG Peng;QIAO Kun;REN Yu-sheng;LIANG Chun;LENG Bing;WU Zong-gui;Department of Cardiology,PLA 273 Hospital;Department of Psychology,Lanzhou University;Department of Cardiology,Changzheng Hospital,Second Military Medical University;

【机构】 解放军第273医院心内科； 兰州大学心理学教研室； 第二军医大学附属长征医院心内科；

【摘要】 目的:水飞蓟素对雷帕霉素诱导的内皮祖细胞(EPCs)凋亡和生长抑制的拮抗作用。方法:通过密度梯度离心法分离、培养、鉴定得到EPCs;在EPCs培养物中加入不同浓度(0、0.1、1、10、100 ng/ml)的雷帕霉素干预24 h和加入(1 ng/ml)雷帕霉素干预不同时间(0、6、12、24、48 h)。收集细胞检测其增殖和迁移能力。在EPCs培养物中加入不同浓度(0、25、50、100μg/ml)的水飞蓟素干预24 h后,收集细胞检测其增殖、迁移和凋亡的水平。在EPCs培养物中同时加入(1 ng/ml)雷帕霉素和不同浓度(25、50、100μg/ml)水飞蓟素干预24 h后,收集细胞检测其增殖、迁移、凋亡和体外血管生成的能力。结果:与未药物干预的EPCs培养物相比较,雷帕霉素(1、10、100 ng/ml)能抑制EPCs增殖和迁移,并随干预浓度的增加和干预时间的延长而增加(P<0.05)。水飞蓟素干预24 h后,50μg/ml和100μg/ml的水飞蓟素可明显增加EPCs增殖和迁移的能力(P<0.05),25~100μg/ml的水飞蓟素司明显抑制EPCs凋亡(P<0.05),并呈浓度依赖性。EPCs在加入雷帕霉素(1 ng/ml)和不同浓度水飞蓟素(25、50、100μg/ml)共同干预24 h后,50μg/ml和100μg/ml的水飞蓟素都可明显改善雷帕霉素对EPCs的促凋亡作用,并能逆转雷帕霉素对EPCs增殖、迁移和血管形成能力的抑制作用(P<0.05)。结论:雷帕霉素能抑制EPCs增殖和迁移,并呈浓度和时间依赖性。水飞蓟素能增强EPCs增殖和迁移,抑制其凋亡,并呈浓度依赖性。水飞蓟素能抑制雷帕霉素对EPCs的促凋亡作用,并能逆转雷帕霉素对EPCs增殖、迁移和血管形成能力的抑制作用。更多还原

【Abstract】 AIM:To study the effect of rapamycin on proliferation and migration of endothelial progenitor cells(EPCs) and to observe the protective effect of silymarin on EPCs under rapamycin intervention.METHODS:Peripheral blood was collected from human volunteers.Mononuclear cells(MNCs) were separated by density centrifugation and were induced to differentiate into EPCs in vitro.EPCs were intervened with different concentrations of rapamycin(0,0.1,1,10,100 ng/ml) for 24 h and different times(0,6,12,24,48 h) and proliferation and migration of EPCs were then detected.EPCs were intervened with different concentrations of silymarin(0,25,50,100 μg/ml) for 24 h and proliferation,migration and apoptosis of EPCs were then detected.Rapamycin(1 ng/ml) and silymarin(25,50,100μg/ml) were added in EPCs for 24 h and proliferation, migration, apoptosis and angiogenesis were then detected.RESULTS:Compared with those in the control group,rapamycin(1,10,100 ng/ml) inhibited proliferation and migration of EPCs in a concentration- and time-dependent manner(P < 0.05).Silymarin(50,100 μg/ml) enhanced proliferation and migration of EPCs and inhibition of apoptosis in a concentration-dependent manner(P <0.05).After adding rapamycin(1 ng/ml) and silymarin(25,50,100 μg/ml) for 24 h,silymarin inhibited the pro-apoptotic effect of rapamycin on EPCs and reversed the inhibitive effect of rapamycin on proliferation,migration and angiogenesis of EPCs(P < 0.05).CONCLUSION:Rapamycin inhibits proliferation and migration of EPCs in a concentration- and time-dependent manner.Silymarin enhances proliferation and migration of EPCs and inhibition of apoptosis in a concentrationdependent manner.Silymarin also inhibits the pro-apoptotic effect of rapamycin on EPCs and reverses the inhibitive effect of rapamycin on proliferation,migration and angiogenesis of EPCs.更多还原