不同浓度Netrin-1对人血管内皮祖细胞增殖凋亡的影响  

Effect of different concentrations of Netrin-1 on proliferation and apoptosis in endothelial progenitor cells

作　　者：艾玲[1] 彭忠英[1] 钟少平[1] 

机构地区：[1]浙江省嘉兴市妇幼保健院产科,314015

出　　处：《中华全科医学》2015年第5期713-715,F0003,共4页Chinese Journal of General Practice

基　　金：浙江省嘉兴市科技局基金资助项目(2012AY1073-1;2012AY1073-5;2012AY1073-6;2013AY21050-5);2012年浙江省医药卫生科技计划项目(2012KYB209)

摘　　要：目的观察不同浓度Netrin-1对人血管内皮祖细胞(Endothelial progenitor cells,EPCs)增殖凋亡的调节作用,探究其在胎盘形成过程中的作用。方法对EPCs进行培养,以递增浓度的Netrin-1(0、0.1、1、10、100 ng/ml)预处理细胞24 h后,应用噻唑兰(MTT)比色法、吖啶橙/溴化乙啶(AB/EO)染色法检测经不同浓度的Netrin-1预处理细胞24 h后EPCs的凋亡情况,采用实时定量PCR及western印迹分别检测Netrin-1mRNA及蛋白变化情况。结果在培养的EPCs中加入不同剂量的Netrin-1干预EPCs 24 h后,EPCs Netrin-1mRNA表达量均升高,但其表达量与Netrin-1的干预剂量无明显量效关系(P>0.05)。经不同浓度Netrin-1干预后,EPCs Netrin-1蛋白表达水平未见明显变化。联合MTT及AB/EO检测表明Netrin-1对EPCs细胞增殖无影响,但EPCs细胞凋亡率随Netrin-1浓度增加而降低,当Netrin-1刺激浓度达100 ng/ml时,EPCs总凋亡率下降至8.26%,较空白组(0 ng/ml)86.12%凋亡率下降明显(P<0.01)。结论 Netrin-1能降低EPCs细胞凋亡率,Netrin-1可能通过影响EPCs凋亡参与胎盘形成过程,起到维持妊娠的作用。Objective To explore the role of different dose of Netrin-1 on regulating the proliferation and apoptosis in endothelial progenitor cells（ EPCs）,and explore the probable function during placentation. Methods EPCs were cultured and treated with different doses of neogenin（ 0 ng / ml,0. 1 ng / ml,1 ng / ml,10 ng / ml,100 ng / ml） for 24 h. Cell viability was measured by MTT assay. EPCs cell apoptosis were assessed by Acridine orange / ethidium bromide（ AB / EO） staining. And a study of Netrin-1 expression in EPCs was performed by Real-time PCR and Western blot. Results Immediately after 24 hour of treatment with Netrin-1,the expression amount of Netrin-1 mRNA in EPCs increased with the accrescence of concentration,but there were no significant dose effect relationship between the expression amount and the intervention dose of Netrin-1（ P〉0. 05）. But the change of Netrin-1 protein in EPCs was seen obvious change by using Western blot. Furthermore,the result of MTT and AB / EO staining showed that proliferation of EPCs was independent of Netrin-1. Meanwhile,cell apoptosis decreased with the accrescence of concentration. The apoptosis rate significantly increased to 8. 26% at 100 ng / ml Netrin-1 and 86. 12% without Netrin-1（ P〈0. 01）. Conclusion Netrin-1 can reduce cell apoptosis in EPCs,Netrin-1 may play a important role in placentation with regulating cell apoptosis of EPCs then maintain pregnancy.

关 键 词：NETRIN-1 血管内皮祖细胞 增殖 细胞凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R714[医药卫生—基础医学]