Am80抑制血管内皮细胞增殖和血管新生内膜形成的机制研究  

Inhibitory effects of Am80 on proliferation in vascular endothelial cells and neointima hyperplasia of carotid arteries

作　　者：王莹[1] 吕心瑞[2] 

机构地区：[1]河南大学第一附属医院,河南开封475004 [2]河南大学医学院生理教研室,河南开封475004

出　　处：《中国病理生理杂志》2016年第12期2205-2210,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.U1404310;No.81600940);河南大学博士启动基金(No.B2013043)

摘　　要：目的:探讨维甲酸衍生物Am80抑制血管内皮细胞增殖和大鼠颈总动脉内皮剥脱术后新生内膜增生的机制。方法:用不同浓度Am80处理EA-hy926细胞24 h后,应用细胞计数和MTS细胞活力分析检测细胞的增殖情况;将Am80处理的细胞进行PI染色,用流式细胞术检测细胞周期的变化情况;通过real-time PCR方法分析Am80处理后EA-hy926细胞中cyclin B1、P21和基质金属蛋白酶(MMP)-2表达的变化;制备SD大鼠颈总动脉内皮球囊损伤模型,通过HE和免疫组化染色方法,观察Am80对球囊损伤后新生内膜形成的影响。结果:细胞计数和MTS细胞活力分析结果显示,Am80以浓度依赖性方式抑制EA-hy926血管内皮细胞增殖;流式细胞分析显示,Am80可使EA-hy926细胞周期停滞在G2/S期;real-time PCR结果表明,Am80处理后,EA-hy926细胞中P21表达上调,cyclin B1和MMP-2表达水平降低;内皮损伤动物模型结果显示,Am80处理后新生内膜的形成受到显著抑制。结论:Am80可通过促进P21而抑制cyclin B1表达,从而抑制血管内皮细胞增殖和血管新生内膜的形成。AIM：To explore the inhibitory effects of Am80 on the proliferation in the vascular endothelial cells（ VECs） and neointima hyperplasia of the carotid arteries after balloon injury in the rats.METHODS：The proliferation of EA-hy926 cells were detected by cell counting and MTS assay after the cells were treated with various doses of Am80 for 24 h.The cell cycle was analyzed by flow cytometry after the cells were stained with PI.The mRNA expression of cyclin B1,P21 and matrix metalloproteinase（ MMP）-2 in the EA-Hy926 cells was detected by real-time PCR.The changes of neointima hyperplasia in the carotid arteries were observed under microscope with hemotoxylin and eosin（ HE） staining,and the expression of cyclin B1 was examined by the method of immunohistochemistry.RESULTS：The proliferation of EA-Hy926 cells was obviously inhibited in a dose-dependent manner when the cells were treated with various doses of Am80 for 24 h.The cell cycle was arrested at G2/S stage in response to Am80 treatment.The mRNA expression of P21 was increased,however,the mRNA expression of cyclin B1 and MMP-2 was decreased when the cells were treated with Am80 at 4 μmol/L for various times.In addition,the vivo experiment demonstrated that Am80 not only significantly reduced neointimal hyperplasia and the thickness ratio of intima to tunicae media compared with injured group,but also inhibited cyclin B1 expression in the carotid arteries.CONCLUSION：Am80 inhibits the proliferation of VECs and neointima hyperplasia in the carotid arteries after balloon injury by promoting P21 expression and decreasing cyclin B1 expression.

关 键 词：Am80 血管内皮细胞 内膜增生 细胞周期 

分 类 号：R363.2[医药卫生—病理学]