平阳霉素诱导人脐静脉血管内皮细胞HUVEC凋亡的作用机制  

Apoptotic Relevant Mechanics of Human Umbilical Vein Endothelial Cells Induced by Pingyangmycin

作　　者：佟笑[1] 夏思文[2] 康宗辉[2] 胡献惠[2] 薛向阳[3] 黄益灯[1] 

机构地区：[1]温州医科大学第一临床学院,325000 [2]中国人民解放军第118医院耳鼻咽喉头颈外科,温州325000 [3]温州医科大学分子病毒与免疫研究所,325000

出　　处：《医学研究杂志》2016年第5期86-90,共5页Journal of Medical Research

基　　金：全军十二五基金资助项目(面上项目)(CWS1IJ256);南京军区重大专项基金资助项目(15ZD010)

摘　　要：目的检测不同浓度平阳霉素(pingyangmycin,PYM)作用24h对人脐静脉血管内皮细胞HUVEC细胞的生长抑制及凋亡的影响,探讨平阳霉素诱导HUVEC细胞凋亡最佳浓度及有关作用机制。方法用不同浓度的平阳霉素作用HUVEC细胞24h,以细胞增殖-毒性8检测试剂盒(cell counting kit-8,CCK-8)检测比较0.1、10、100、1000μg/ml平阳霉素作用24h后对HUVEC细胞的抑制作用,用膜联蛋白V-异硫氰酸荧光素/碘化丙啶细胞凋亡检测试剂盒(Annexin-V FITC/PI apoptosis kit)检测不同浓度PYM对HUVEC细胞凋亡作用的影响,蛋白印迹法(Western blot)检测不同浓度平阳霉素作用HUVEC细胞后内质网应激蛋白GRP78和CHOP的表达。结果 CCK-8结果显示随平阳霉素浓度的升高,对HUVEC细胞的抑制作用越明显(P<0.05);然平阳霉素药物浓度100μg/ml和1000μg/ml组相比,差异无显著性统计学意义(P>0.05)。流式细胞检测显示0.1、1、10、100μg/ml平阳霉素作用24h后细胞凋亡率分别为5.37%±1.63%、12.70%±1.57%、24.70%±6.99%、23.60%±0.80%,坏死率分别为4.87%±2.09%、7.32%±3.88%、39.10%±6.18%、66.30%±0.40%,随药物浓度增加而增高,过高浓度细胞坏死占主导,和对照组相比,差异具有统计学意义(P<0.05);Western blot法结果显示,平阳霉素各浓度作用24h后促进内质网应激蛋白GRP78和CHOP的表达。结论平阳霉素在体外能明显抑制HUVEC细胞生长并促进其凋亡,当平阳霉素浓度达到100μg/ml时,细胞以坏死为主;平阳霉素可能通过内质网应激(endoplasmic reticulum stress,ERS)途径诱导HUVEC细胞凋亡。Objective To investigate the viability and apoptosis effect of Pingyangmycin （ PYM ） on cultured human umbilical vein endothelial cells（HUVEC） in vitro and the possible mechanism. Methods CCK -8 reagents was performed to detect the inhibition of HUVEC with 0.1,1,10,100,1000μg/ml of PYM for 24h. The apoptosis and necrosis of HUVEC were detected through flow Cytometry. Western blot assay were used to determine GRP78 ,CHOP by 0.1,1,10,100μg/ml of PYM for 24h. Results CCK - 8 reagents showed that HUVEC were inhibited more significantly with increasing PYM concentration （ P 〈 0.05 ）. Annexin V and propidium iodide （PI） stainning show that the apoptosis rates of HUVEC induced by 0.1,1,10,100μg/ml of PYM for 24h were 5.37% ± 1.63% , 12.70% ± 1.57% ,24.70%±6.99% ,23.60%±0.80% , necrosis rate were 4.87% ± 2.09% ,7.32% ± 3.88% ,39.10% ± 6.18% .66.30%±0. 40%. The higher concentration may lead necrosis of cells. Western blot analysis showed that PYM prompted Endoplasmic reticulum stress proteins GRP78,CFIOP express. Conclusion PYM can inhibit HUVEC cells in vitro effectively in a concentration - dependent manner. The reason that PYM induce the apoptosis of HUVEC cells might be through endoplasmic reticulum stress.

关 键 词：血管瘤 平阳霉素 糖调节蛋白78 C/EBP同源蛋白 内质网应激 

分 类 号：R762[医药卫生—耳鼻咽喉科] R979.14[医药卫生—临床医学]