体外震波对人脐静脉内皮细胞增殖、细胞周期及细胞间黏附因子-1表达的影响  

Effect of Extracorporeal Shock Wave on Proliferation,Cell Cycle and Intercellular Adhesion Molecule-1 Expression in Human Umbilical Vein Endothelial Cells

作　　者：马一铭 李丽[1] 蔡红雁[1] 胡钊[1] 郭涛[1] 

机构地区：[1]昆明医科大学第一附属医院心内科,云南省昆明市650000

出　　处：《中国循环杂志》2016年第10期1013-1017,共5页Chinese Circulation Journal

基　　金：国家自然科学基金项目(81260027);云南省卫生厅科技计划项目(2012WS005);云南省科技计划项目(2014FZ023);云南省科技厅-昆明医科大学联合专项基金(2013FB128)

摘　　要：目的:观察体外震波治疗对人脐静脉内皮细胞(HUVECs)增殖、细胞周期及细胞间黏附因子-1(ICAM-1)表达的影响。方法:将体外培养的人脐静脉内皮细胞分组,实验部分分别给予0.03、0.09、0.18、0.24 m J/mm2能量震波500击处理(分别设定为0.03、0.09、0.18、0.24 m J/mm^2能量组),对照组不给予震波处理。细胞计数试剂盒(CCK)比色法检测细胞增殖情况,流式细胞术检测细胞周期,实时定量聚合酶链反应(PCR)及蛋白质印迹方法(Westen blot)分别检测低能量震波处理后ICAM-1信使核糖核酸(m RNA)和蛋白的表达水平。结果:CCK比色法检测结果显示:只有0.09 m J/mm2能量组与对照组比能促进HUVECs增殖,且差异有统计学意义(P<0.05),而其他能量组与对照组比,差异均无统计学意义(P>0.05)。不同能量组与对照组比较,0.09 m J/mm^2能量组G0/G1期细胞比例明显减少(P<0.05),S期和G2/M期细胞比例明显增加(P<0.05),0.03 m J/mm2能量组仅增加G2/M期细胞比例(P<0.05),而其他能量组与对照组比差异均无统计学意义(P>0.05)。实时定量PCR检测显示:0.09m J/mm^2能量组ICAM-1 m RNA表达均较对照组明显增高(9.27±0.95 vs 1.02±0.27,P<0.001),0.03 m J/mm2能量组ICAM-1 m RNA表达均较对照组亦增高(7.08±0.60 vs 1.02±0.27,P<0.01)。Westen blot检测显示:0.09 m J/mm^2能量组ICAM-1蛋白表达较对照组明显增高,差异有统计学意义(P<0.05)。结论:体外震波治疗,特别是0.09 m J/mm^2能量的体外震波能加速HUVECs细胞周期由G0期/G1期向S期和G2/M期的转换,促进细胞增殖,并且提高ICAM-1的表达,这在体外震波促进血管新生机制中起重要作用。Objective：To observe the effect of extracorporeal shock wave therapy（ESWT） on proliferation,cell cycle and intercellular adhesion molecule-1（ICAM-1） expression in human umbilical vein endothelial cells（HUVECs）.Methods：HUVECs were cultured in vitro at the concentration of（1×105/ml） and the cells were divided into 2 sets of groups：CSWT group,the cells were treated by different energy of（0.03,0.09,0.18,0.24） m J/mm2 respectively and corresponding Control group,in which the cells had no CSWT.HUVEC proliferation was detected by CCK colorimetric method,cell cycle was measured by flow cytometry,m RNA and protein expressions of ICAM-1 were examined by RT-PCR and Western blot analysis respectively.Results：Compared with Control group,（0.09 m J/mm2） CSWT group had promoted HUVECs proliferation,P0.05 and the other CSWT groups were similar to corresponding Control groups,P0.05;（0.09 m J/mm2） CSWT group showed decreasedproportion of G0/G1 stage and increased S and G2/M stages,all P0.05;while（0.03 m J/mm2） CSWT group only increased the proportion of G2/M stage,P0.05 and the other CSWT groups were similar to corresponding Control group,P0.05.Compared with Control group,（0.09 m J/mm2） and（0.03 m J/mm2） CSWT groups showed increased m RNA expression of ICAM-1（9.27±0.95） vs（1.02±0.27）,P0.001 and（7.08±0.60） vs（1.02±0.27）,P0.01;（0.09 m J /mm2） CSWT group had elevated protein expression of ICAM-1,P0.05.Conclusion：ESWT especially at（0.09 m J/mm2） may accelerate cell cycle transition from G0/G1 stage to S and G2/M stages,promote HUVECs proliferation and increase ICAM-1 expression which may play important roles in ESWT facilitated angiogenesis in vitro.

关 键 词：体外震波治疗 脐静脉 内皮细胞 细胞周期 

分 类 号：R54[医药卫生—心血管疾病]