蚤休皂苷对氧化损伤脐静脉内皮细胞内钙离子变化的影响_药学论文.docVIP

蚤休皂苷对氧化损伤脐静脉内皮细胞内钙离子变化的影响_药学论文 蚤休皂苷对氧化损伤脐静脉内皮细胞内钙离子变化的影响_药学论文 作者：高琳琳，李福荣，康莉，司艳红，王浩 【摘要】 　　目的利用激光共聚焦显微镜研究蚤休皂苷（Pa）对H2O2诱导的脐静脉内皮细胞（ECV304细胞）损伤的钙离子（Ca2+）的变化及其机制。方法体外培养ECV304细胞，建立氧化损伤的细胞模型，然后分为5个实验处理组：①正常对照组；②氧化损伤组；③高浓度蚤休皂苷（1 μg/ml）组 ；④中浓度蚤休皂苷（100 pg/ml）组；⑤低浓度蚤休皂苷（10 pg/ml）组；应用激光共聚焦显微镜观察其细胞凋亡的形态学差异。并利用激光共聚焦显微镜观察、图像分析仪分析各实验分组细胞中游离Ca2+的表达、激光共聚焦显微镜描记预处理前后Ca2+的动态表达。结果损伤组细胞内游离Ca2+浓度明显高于正常组(0.01)，而各蚤休皂苷预处理组均使Ca2+荧光强度下降，显著低于损伤组(0.01)，并呈剂量依赖性效应。H2O2损伤剂量的刺激下，ECV304胞浆内的游离Ca2+呈现双相变化：由静息状态持续增加到峰值，ΔF为37.22±3.72，平台期与静息状态荧光值的变化ΔP为7.65±0.69，加入蚤休皂苷预处理10 min后，再加入100 μmol/L的H2O2 ，ΔF和ΔP均明显下降(0.01)。结论蚤休皂苷可以保护H2O2造成的ECV304细胞的氧化损伤，与抑制了钙离子介导的凋亡通路有关，达到保护内皮细胞、抗动脉粥样硬化的目的。 【关键词】 蚤休皂苷 内皮细胞 凋亡 钙离子 动脉粥样硬化 　　Abstract：ObjectiveTo investigate the effects and mechanisms of Pariphyllin （Pa）on calcium ion density in injury ecv304 induced by hydrogen peroxide.MethodsECV304 cells were cultured in vitro, the oxidative injury was built by hydrogen peroxide (H2O2）,cells were divided into five experimental groups:1.normal,2.oxidative damage group,3.high density Pariphyllin（1 μg/ml）,4.medium density Pariphyllin （100 pg/ml）and 5.low density Pariphyllin（10pg/ml）.Laser confocal microscopy(LCM) was used to observe quiet state and dynamic change of the density of calcium ion (Ca2+).Results The LCM showed that in damaged group, the external Ca2+ concentation was the highest,but in pretreated groups,it was decreased in dose dependent manner（0.01）.H2O2 elicited an transient peak of ［Ca2+］i and then fell to a subsequent sustained higher plateau.Pretreatment with pariphyllin（1 μg/ml）for 10 minute significantly prevented the transient increase and sustained the increase of ［Ca2+］i.ConclusionPariphyllin has protective action against oxidative damage of ECV304 induced by H2O2, its mechanism may be related to decreasing the cell apoptosis and inhibiting calcium ion-decreased mediated signal transduction . 　　Key words： Pariphyllin; HUVEC; Apoptosis; Ca2+; Atherosclerosis 　　内皮细胞损伤是动脉粥样硬化(Atherosclerosis,AS)发生的关键环节，目前占主导地位的“损伤-