超声联合微泡增加内皮细胞中内皮型一氧化氮合酶和一氧化氮生成的体外实验

目的 评估诊断性超声联合微泡对内皮细胞中内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及一氧化氮(nitric oxide,NO)生成的增强效应.方法 将正常培养的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)分为空白对照组(A组)、纯微泡组(B组)、单纯超声组(C组)和超声联合微泡组(D组).其中,D组按条件不同又分为:辐照时间不同组(1 min、5 min、10 min)、机械指数不同组(0.09、0.4、1.0)、微泡浓度不同组(5×10~8/ml、2.5×10~8/ml、1.25×10~8/ml).干预后即刻和干预后24 h分别在光镜下观察细胞形态结构,RT-PCR测细胞eNOS相对表达量,NO试剂盒测培养液中NO水平,并用统计学方法对上述指标进行组间比较.结果 D组中eNOS及NO含量明显较其他三组高;当参数选择为机械指数1.0、微泡浓度2.5×10~8/ml、辐照时间10 min时,D组中eNOS和NO的增加程度最明显;并且干预后即刻和干预后24 h各组细胞及细胞膜形态结构在光镜下均无显著变化.结论 超声联合微泡辐照能增加内皮细胞中eNOS和NO生成;给予的超声及微泡条件不同,其增强效果也不同.

Ultrasound combined with microbubble enhanced eNOS expression and NO release in endothelial cells in vitro

Objective To assess the enhancement effect of diagnostic ultrasound combined with microbubbles in the generation of endothelial nitric oxide synthase(eNOS)and nitric Oxide(NO)in endothelial cells.Methods Normal cultured human umbilical vein endothelial cells(HUVEC)were divided into blank control group(A group),simple microbubble group(B group),simple ultrasound group(C group)and ultrasound combined with microbubble group(D group).According to different conditions,group D was divided into three sub-groups:different time groups(1 min,5 min,10 min);different machinery index(MI)groups(0.09,0.4,1.0),different microbubble concentration groups(5×10~8/ml,2.5×10~8/ml,1.25×10~8/ml).Cell morpha was observed in the light microscope immediately and 24 h after the intervention,respectively.RT-PCR was used to measure the relative expression of eNOS in cells.NO kit was used to measure the NO Ievels in culture medium.And statistical methods were used to analyse the experimental data.Results NO and eNOS were significantly higher in group D than the other three groups.When MI=1.0,microbubble concentration=2.5×10~8/ml,and irradiation time=10 min,the increase of eNOS and NO in group D was the most obvious.Furthermore,the cell morphology had no significant change in the light microscope immediately and 24 h after the intervention.Conclusions Ultrasound combined with microbubble can increase the generation of eNOS and NO in endothelial cells.