祛痰、化瘀和祛痰化瘀方对氧化低密度脂蛋白诱导血管内皮细胞增殖及凋亡功能的影响

摘要

目的:观察祛痰、化瘀和祛痰化瘀方对ox-LDL损伤HUVEC增殖及凋亡功能的影响.方法:建立HUVEC ox-LDL损伤模型,采用MTT检测法、AnnexinV-FITC/P1双染结合流式细胞术和Western blot检测法,观察方药对HUVEC增殖率、凋亡率及与增殖、凋亡相关蛋白VEGFR、P66、P53、Bcl-2、BAX表达的影响.结果:在细胞增殖方面,祛痰化瘀方能增加HUVEC吸光度,祛痰方、祛痰化瘀方可上调HUVEC VEGFR的表达水平,ox-LDL作用下4种方剂均可增加HUVEC吸光度并上调VEGFR表达水平.在细胞凋亡方面,祛痰方、化瘀方和祛痰化瘀方均能降低P66的表达水平;ox-LDL作用下,HUVEC Bcl-2表达降低,BAX、P66表达升高;祛痰化瘀方等4种方剂对早期、晚期及总凋亡率都有极大改善,且均可降低BAX的表达水平;祛痰方组、化瘀方组和祛痰化瘀方可降低P66的表达水平,祛痰化瘀方可提高Bcl-2的表达水平,Ox-LDL及4种方剂均对P53的表达水平无明显影响.结论:祛痰、化瘀和祛痰化瘀方及基础方均可在不同程度上通过促进增殖、抗凋亡达到对抗HUVEC氧化损伤的作用,其中祛痰化瘀方的效果最显著.%Objective:To observe the effect of Qutan-Fang,Huayu-Fang,and Qutan-Huayu-Fang on proliferation and apoptosis of Human umbilical vascular endothelial cells induced by ox-LDL.(HUVECs) in oxidative injury.Methods:HUVEC were divided into normal group,model group,Basic-Fang group,Qutan-Fang group,Huayu-Fang group and Qutan-Huayu-Fang group,which treated with ox-LDL(100mg/ml).Proliferation of HUVECs were detected by MTT.Apoptosis of HUVECs were analysed by AnnexinV-FITC/PI staining with flow cytometry.And the expression of VEGFR,P66,P53,Bcl-2,BAX were detected by Western blot.Results:Compared with normal group,Qutan-Huayu-Fang can promote the light absorption value of HUVEC,and Qutan-Fang,Qutan-Huayu-Fang can increase the expression of VEGFR.All four prescriptions can increase the light absorption value on HUVEC and raising the expression of VEGFR stimulated with ox-LDL.Compared with normal group,Qutan-Fang,Huayu-Fang,Qutan-Huayu-Fang can reduce the expression of P66 of HUVEC.Stimulated with ox-LDL,the expression of Bcl-2 was reduced,and the expression of BAX 、P66 were increased.All four prescriptions can improve the early apoptotic rate,the late apoptotic rate,and the total apoptotic rate.These four prescriptions can reduced the expression of BAX.Qutan-Fang,Huayu-Fang,Qutan-Huayu-Fang can reduce the expression of P66 stimulated with ox-LDL.Qutan-Huayu-Fang can increase the expression of Bcl-2 of HUVEC.Conclusion:Qutan-Fang,Huayu-Fang,Qutan-Huayu-Fang can effectively reduce the oxidative injury on HUVECs by improve their proliferation and inhibiting their apoptosis.The effect of Qutan-Huayu-Fang was the most significant in all four prescriptions.