人脐静脉内皮细胞衰老过程中细胞膜钠泵α亚单位基因表达的变化  

Expression of cell membrane sodium pumpαsubunit genes in aging of human unbilical vein endothelial cells

作　　者：张亭[1] 任延平[1] 张明娟[2] 黄若文[1] 

机构地区：[1]西安交通大学第一附属医院老年心内科,710061 [2]西安交通大学第二附属医院心内科

出　　处：《中华老年心脑血管病杂志》2012年第9期971-976,共6页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(30971222);陕西省科技攻关项目(2008K13-01)

摘　　要：目的探讨体外培养的人脐静脉内皮细胞(HUVEC)衰老过程中,细胞膜表面钠泵α亚单位各异构体基因表达的改变。方法提取原代HUVEC,体外传代培养,第1代起分组干预。实验分对照组(未干预)和干预组(D-半乳糖10 g/L干预),按细胞传代的不同代数,观察第2、5和8代细胞形态,于不同群体倍增数(PD)行β-半乳糖苷酶染色初步判定细胞衰老,利用免疫细胞荧光染色法、实时定量PCR法和Western b1ot法检测各组细胞膜表面钠泵α1、α2、和α3亚单位mRNA及蛋白的表达水平。结果钠泵α1、α2和α3亚单位mRNA及蛋白表达水平随PD增加而降低(P<0.01);与对照组比较,干预组同一PD钠泵α2和α3亚单位mRNA和蛋白表达降低(P<0.01),而2组α1亚单位表达差异无统计学意义(P>0.05)。结论细胞膜表面钠泵α亚单位mRNA及蛋白表达水平的失衡,可能是HUVEC复制性衰老的分子机制之一;D-半乳糖诱导可能构建HUVEC亚急性衰老模型。Objective To study the expression of cell membrane sodium pump α subunit is genes in aging of in vitro cultured HUVEC. Methods Extracted and in vitro subcultured pri HUVEC were divided into non galactose treatment （control group） and galactose treat group. Morphology of the second, fifth and eighth generations of HUVEC was observed. Agi HUVEC was identified according to their population doublings（PD） with β-galactosidase stai- ning. Expression of cell membrane sodium pump subunits mRNA and protein was detected in two groups by RT-PCR and Western blot respectively with immunofluorescence staining. Results The expression levels of cell membrane sodium pump α1, α2 and α3 subunits mRNA and protein decreased with the increasing PD（P〈0.01）. The expression levels of cell membrane sodium pump α2 and α3 units mRNA and protein were significantly lower in galactose treatment group than in control group（P〈0.01）, No significant difference was found in cell membrane sodium pump α1 subunit expression level between the two groups（P〉0.05）. Conclusion Unbalanced expression of cell membrane sodium pump α subunits mRNA and protein may be one of the molecular mecha- nisms underlying the aging of HUVEC. D-galactose can induce subacue aging model of HUVEC.

关 键 词：脐静脉 内皮细胞 细胞衰老 细胞膜 RNA 信使 钠钾交换ATP酶 基因表达 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]