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[人胚肺成纤维细胞复制性衰老和早老过程中衰老相关基因表达及端粒酶活性的变化]

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

作者信息

Yang J P, Zhang W J, Jing C X, Wu C P, Ji W D, Yang L Q, Zhuang Z X

机构信息

Occupational health branch, Shenzhen Taike detection Co. Ltd , Shenzhen 518074, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2017 Jan 20;35(1):15-18. doi: 10.3760/cma.j.issn.1001-9391.2017.01.004.

Abstract

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. The HELFs were divided into young cells (22 population doubling levels, 22PDL) , mid-aged cells (35PDL) and replicative senes-cent cells (49PDL) and premature senescent cells induced by H(2)O(2)(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 Foxo1, Foxo3, Pdx1, apoA-I and MMP1 was per-formed by RT-Q-PCR separately. The mRNA level for Foxo1, Foxo3, apoA-I and Pdx1 was decreased separately during cellular replicative senescence compared to that in the young-stage cells with statistical signifi-cance (<0.05). The expression of MMP1 was up-regulated 5.1-fold obviously (<0.05). In premature senes-cence, the mRNA level was only decreased for Foxo1, Foxo3 and apoA-I, but up-regulated 2.3-fold and 6.2-fold for Pdx1 and MMP1 respcetively vs 22PDL significantly (<0.05). The telomerase activity in young cells was not detected, and it increased in mid-aged cells and replicative senescence stages during cellular replicative se-nescence as compared to 22PDL with statistical significance (<0.05). The telomerase activity in premature se-nescence was highly active. The expression for genes related with senescence has differences be-tween replicative and premature senescence and hydrogen peroxide modifies their expression levels. The telomer-ase activity has been going up with increased PDLs.

摘要

检测人胚肺成纤维细胞(HELFs)在体外细胞复制性衰老和过氧化氢诱导的早衰过程中端粒酶活性的变化以及与衰老相关的氧化应激反应基因的表达。将HELFs分为年轻细胞(22个群体倍增水平,22PDL)、中年细胞(35PDL)、复制性衰老细胞(49PDL)以及H₂O₂诱导的早衰细胞(早衰,PS)。在细胞复制性衰老和早衰过程中,通过ELISA法检测端粒酶活性。分别通过RT-Q-PCR检测与衰老相关的氧化应激反应基因Foxo1、Foxo3、Pdx1、载脂蛋白A-I和基质金属蛋白酶1(MMP1)的mRNA水平。与年轻阶段细胞相比,细胞复制性衰老过程中Foxo1、Foxo3、载脂蛋白A-I和Pdx1的mRNA水平分别显著降低(<0.05)。MMP1的表达明显上调5.1倍(<0.05)。在早衰过程中,仅Foxo1、Foxo3和载脂蛋白A-I的mRNA水平降低,而Pdx1和MMP1的mRNA水平分别比22PDL显著上调2.3倍和6.2倍(<0.05)。年轻细胞中未检测到端粒酶活性,在细胞复制性衰老过程中,中年细胞和复制性衰老阶段的端粒酶活性与22PDL相比增加,具有统计学意义(<0.05)。早衰过程中端粒酶活性高度活跃。与衰老相关的基因表达在复制性衰老和早衰之间存在差异,过氧化氢会改变它们的表达水平。端粒酶活性随群体倍增水平的增加而升高。

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