平阳霉素诱导人血管瘤内皮细胞凋亡过程中PARP-1和Cyt-C的表达及意义  

The expression and significance of PARP-1 and Cyt-C in the process of pingyangmycin inducing hemangioma endothelial cell to apoptosis

作　　者：葛平[1] 蒋菲[2] 屠军波[2] 虎小毅[2] 马瑞朝[2] 贺琳英[1] 杨壮群[1] 

机构地区：[1]西安交通大学第一附属医院烧伤整形科,陕西西安710061 [2]西安交通大学附属口腔医院口腔颌面外科

出　　处：《中国美容医学》2012年第10期1760-1764,共5页Chinese Journal of Aesthetic Medicine

基　　金：2011年度陕西省科技计划科研项目(2011K14-01-06);2007年国家自然科学基金(30700954)

摘　　要：目的:探讨平阳霉素诱导人血管瘤内皮细胞凋亡过程中PARP-1、Cyt-C的表达及可能机制。方法:在体外培养人血管瘤内皮细胞,设立两组:A组为平阳霉素组,其中加入平阳霉素(200μg/ml),诱导血管瘤内皮细胞凋亡,B组为空白对照组。对两组血管瘤内皮细胞通过倒置显微镜、电镜观察其形态学变化,采用流式细胞仪检测平阳霉素诱导血管瘤内皮细胞的凋亡率。利用基因芯片技术筛选出差异表达的基因,从中选定PARP-1及Cyt-C表达基因,对其进行RT-PCR验证。结果:平阳霉素(200μg/ml)可以诱导体外培养的人血管瘤内皮细胞凋亡,加药24h后,在倒置显微镜和电镜下观察到明显的细胞凋亡的形态学改变,流式细胞仪检测凋亡率可达13.11%。空白对照组血管瘤内皮细胞倒置显微镜下观察无明显形态学变化,流式细胞仪检测凋亡率为3.56%。利用基因芯片技术筛选出两组差异基因4752个,其中2543个上调,2209个下调。应用DAVI D软件对鉴定出来的差异基因进行生物信息学分析,从中选出与线粒体凋亡途径直接相关的蛋白表达基因:PARP-1、Cyt-C。对其进行RT-PCR检测得到的CT值显示:平阳霉素药物组相对于空白组PARP-1及Cyt-C蛋白基因呈高表达状态。结论:平阳霉素在体外可以通过多种途径诱导血管瘤内皮细胞发生凋亡,其中PARP-1及Cyt-C参与了线粒体途径调控的凋亡。Objective To discussing the influence of the PYM inducing Hemangioma endothelial cells to Apoptosis on the expression levels of PARP-1,Cyt-C and its possible mechanism. Methods Culturing Hemangioma Endothelial Cell in Vitro, setting two groups： A group is PYM group, adding PYM（200ug/ml）, to induce Hemangioma Endothelial Cell to apoptosis. B group is the Blank control group. Observe the morphological changes of these two groups using inverted microscope and electron microscope, and detect the apoptosis rate of Hemangioma Endothelial Cell induced by PYM using flow cytometry. Then screen of differentially expressed genes using gene chip technology, and select PARP-1 and Cyt-C among the genes, and do RT-PCR test on them. Results Pingyangmycin （2001Jg/ml） can induce Hemangioma Endothelial cells that cultured in vitro to apoptosis, and 24 hours after dosing, we can find obvious morphological changes of apoptosis, observed via inverted microscope and electron microscope ,and the apoptosis rate is high up to 13.11% detected by flow cytometry. The blank control group on the other side, no obvious morphological changes are observed, and the apoptosis rate is 3.56%. We filter out differentially expressed genes between two groups are 4752 using genes chip technology. Among them ,2543 genes are highly expressed and 2209 are expressed in lower level.Use DAVID sofwareto do Bioinformatic Analysis of the differentially expressed genes which are identified. And select the genes directly related to the the mitochondrial pathway of apoptosis protein genes： PARP-1, Cyt-C. Its CT value obtained from RT-PCR test shows that PARP-1 and Cyt-C protein genes were highly expressed in PYM group comparing to the blank group. Conclusion Through a variety of channels Pingyangmycin in vitro can induce Hemangioma endothelial cell to apoptosis. And PARP-1 and Cyt-C are involved in the regulation of the mitochondrial pathway of apoptosis

关 键 词：血管瘤内皮细胞 凋亡 平阳霉素 PARP-1 Cyt—C 

分 类 号：Q813.1[生物学—生物工程]