过氧化氢诱导细胞早衰中基因表达及端粒酶活力的变化  

Changes of the expression for genes related with senescence and the telomerase activity during cellular replicative and premature senescence in human embryonic lung fibroblasts

作　　者：杨建平 张文娟[2] 荆春霞[2] 吴赤蓬[2] 纪卫东[3] 杨淋清[4] 庄志雄[4] 

机构地区：[1]深圳市太科检测有限公司,518074 [2]暨南大学医学院公共卫生与预防医学系 [3]中山大学第一附属医院转化医学中心 [4]深圳市疾病预防控制中心

出　　处：《中华劳动卫生职业病杂志》2017年第1期15-18,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金（81473014、81001259、81272350、81371217、81273127、81172710）;暨南大学医学院公共卫生与预防医学高水平大学建设项目（JNUPHPM2016003）

摘　　要：目的探讨人胚肺成纤维细胞复制性衰老及过氧化氢诱导早衰过程中，氧化应激应答基因表达谱及端粒酶活力的变化。方法按传代情况将人胚肺成纤维细胞分为年轻细胞（22 population doubling levels，22PDL）组、中年细胞（35PDL）组、复制性衰老细胞（49PDL）组和氧化应激诱导的早衰组Ps。通过荧光定量PCR检测各组细胞中衰老相关的氧化应激应答相关基因（Foxol、Foxo3、Pdxl、apoA-I、MMPl）的mRNA表达水平，分析过氧化氢对基因表达的影响。用ELISA方法检测不同衰老阶段细胞的端粒酶活力变化。结果在人胚肺成纤维细胞复制性衰老过程中，与年轻细胞组比较，Foxol、Foxo3、apoA-I及Pdxl的mRNA表达均逐渐降低，而复制性衰老细胞组MMPl表达明显升高至年轻细胞组的5．1倍，差异有统计学意义（火0．05）；与年轻细胞组比较，过氧化氢诱导的早衰阶段中Foxol、Fox03和apoA-I表达降低，Pdxl和MMPl表达明显升高，分别为年轻细胞组的2．3倍和6．2倍，差异有统计学意义（P〈0．05）。年轻细胞组未检测到端粒酶活力，与年轻细胞组比较，中年及复制性衰老细胞组端粒酶活力增强；过氧化氢诱导的早衰阶段端粒酶活力明显升高，差异均有统计学意义（P〈0．05）。结论人胚肺成纤维细胞复制性衰老与早衰的基因表达存在差异，并受氧化应激修饰；细胞端粒酶活力随增龄呈升高趋势。Objective To detect the alterations of telomerase activity and the expression for oxidative stress responsive genes related with senescence during cellular replicative senescence and hydrogen peroxide- induced premature senescence in human embryonic lung fibroblasts（HELFs） in vitro. Methods The HELFs were divided into young cells （22 population doubling levels, 22PDL）, mid-aged cells （35PDL） and replicative senescent cells （49PDL） and premature senescent cells induced by H2O2 （premature senescence, PS）. The telomerase activity was detected by ELISA assay during cellular replicative and premature senescence. The mRNA level of oxidative stress responsive genes related with senescence for Foxol, Foxo3, Pdxl, apoA-I and MMP1 was performed by RT-Q-PCR separately. Results The mRNA level for Foxol, Foxo3, apoA-I and Pdxl was decreased separately during cellular replicative senescence compared to that in the young-stage cells with statistical significance （P〈0.05）. The expression of MMP1 was up-regulated 5.1-fold obviously （P〈0.05）. In premature senescence, the mRNA level was only decreased for Foxol, Foxo3 and apoA-I, but up-regulated 2.3- fold and 6.2-fold for Pdxl and MMP1 respcetively vs 22PDL significantly （P〈0.05）. The telomerase activity in young ceils was not detected, and it increased in mid-aged cells and replicative senescence stages during cellular replicative senescence as compared to 22PDL with statistical significance （P〈0.05）. The telomerase activity in premature senescence was highly active. Conclusion The expression for genes related with senescence has differences between replicative and premature senescence and hydrogen peroxide modifies their expression levels. The telomerase activity has been going up with increased PDLs.

关 键 词：细胞衰老 过氧化氢 基因表达 

分 类 号：R3416[医药卫生—基础医学]