Tert-butylhydroperoxide induces aging of Sca-1<sup>+</sup> hematopoietic stem and progenitor cells by regulating cell cycle

周玥; 姜蓉; 王萍; 杨斌; 姚欣; 刘典锋; 王亚

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Tert-butylhydroperoxide induces aging of Sca-1⁺ hematopoietic stem and progenitor cells by regulating cell cycle

更新时间：2026-01-09

- Tert-butylhydroperoxide induces aging of Sca-1⁺ hematopoietic stem and progenitor cells by regulating cell cycle
- Journal of Army Medical University Vol. 33, Issue 18, Pages: 1893-1896(2011)
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  CLC： R329.28;R339.38;R914.3
- Published：2011
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周玥,姜蓉,王萍,杨斌,姚欣,刘典锋,王亚.三丁基过氧化氢通过调控细胞周期诱导Sca-1+造血干/祖细胞衰老[J].陆军军医大学学报(原第三军医大学学报),2011,33(18):1893-1896.

　Zhou Yue,Jiang Rong,Wang Ping,Yang Bin,Yao Xin,Liu Dianfeng,Wang Yaping.Tert-butylhydroperoxide induces aging of Sca-1+ hematopoietic stem and progenitor cells by regulating cell cycle[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1893-1896.
周玥,姜蓉,王萍,杨斌,姚欣,刘典锋,王亚.三丁基过氧化氢通过调控细胞周期诱导Sca-1+造血干/祖细胞衰老[J].陆军军医大学学报(原第三军医大学学报),2011,33(18):1893-1896. DOI：

　Zhou Yue,Jiang Rong,Wang Ping,Yang Bin,Yao Xin,Liu Dianfeng,Wang Yaping.Tert-butylhydroperoxide induces aging of Sca-1+ hematopoietic stem and progenitor cells by regulating cell cycle[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1893-1896. DOI：

摘要

目的探讨三丁基过氧化氢（t-BHP）诱导Sca-1

造血干/祖细胞（HSC/HPC）衰老的细胞周期调控机制。方法免疫磁性分选法分离纯化小鼠Sca-1

HSC/HPC后分为对照组（常规培养细胞），衰老模型组（用t-BHP复制细胞体外衰老模型）。通过衰老相关β-半乳糖苷酶（SA-β-Gal）染色、细胞周期测定和造血祖细胞混合集落（CFU-Mix）培养观察t-BHP诱导Sca-1

HSC/HPC衰老的生物学作用； RT-PCR检测衰老相关基因p16

INK4a

、p19

Arf

、p53、p21

Cip1/Waf1

mRNA的表达。Western blot检测细胞周期调控蛋白P16

INK4a

、P21

Cip1/Waf1

、CyclinD1、Cyclin E、CDK4、CDK2的表达。结果免疫磁性分选法分离纯化的Sca-1

HSC/HPC纯度可达（87.33±1.25）%。衰老组细胞SA-β-Gal染色阳性率为（57.9±4.2）% ，高于对照组[（9.1±2.4）%，P0.05

]

，G

期细胞比例为（87.53±4.03）%，高于对照组[（72.26±3.40）%，P0.05

]

，生成造血祖细胞混合集落（CFU-Mix）数为（1.5±1.2）/10

，低于对照组[（9.8±2.1）/10

，P0.05

]

。与对照组比较，衰老组细胞p16

INK4a

、p19

Arf

、p53、p21

Cip1/Waf1

mRNA及P16

INK4a

、P21

Cip1/Waf1

、CyclinD1蛋白表达上调（P0.05），Cyclin E、CDK4、CDK2蛋白表达下调（P0.05）。结论 t-BHP能有效诱导Sca-1

HSC/HPC衰老，其机制可能与调控细胞周期调控分子的表达有关。

Abstract

Objective To study the cell cycle regulation mechanism underlying t

ert-butylhydroperoxide (t-BHP)-induced aging of Sca-1

hematopoietic stem and progenitor cells (HSC/HPC). Methods Sca-1

HSC/HPC were isolated and purified by magnetic activated cell sorting (MACS) and divided into control group and aging model group. Sca-1

HSC/HPC in control group were routinely cultured. An in vitro aging model of Sca-1

HSC/HPC was induced by t-BHP. Biological role of t-BHP in inducing aging of Sca-1

HSC/HPC was observed with aging-associated β-galactosidase (SA-β-gal) staining

cell cycle assay and culture of colony-forming units in mixed HPC. Expressions of aging-related p16

INK4a

p19

Arf

p53

p21

Cip1/Waf1

mRNA and P16

INK4a

P21

Cip1/Waf1

CyclinD1

CyclinE

CDK2 and CDK4 protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting

respectively. Results The purity of Sca-1

HSC/HPC isolated by MACS was 87.33%±1.25% in aging model group. The positive staining rate and the ratio of Sca-1

HSC/HPC were higher in aging model group than in control group (57.9%±4.2% vs 9.1%±4.2%

87.53%±4.03% vs 72.26%±3.4%

P0.05).The number of colony-forming units in mixed HPC was less in aging model group than in control group [(1.5±1.2)/10

vs (9.8±2.1)/10

P0.05

]

. The expression levels of p16

INK4a

p19

Arf

p53

p21

Cip1/Waf1

mRNA

and P16

INK4a

P21

Cip1/Waf1

CyclinD1 protein were higher while those of Cyclin E

CDK4

CDK2 protein were lower in aging model group than in control group（P0.05）. Conclusion t-BHP can effectively induce the aging of Sca-1

HSC/HPC by regulating the expression of cell cycle regulatory molecules.

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Tert-butylhydroperoxide induces aging of Sca-1+ hematopoietic stem and progenitor cells by regulating cell cycle

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摘要

Abstract

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references

Tert-butylhydroperoxide induces aging of Sca-1⁺ hematopoietic stem and progenitor cells by regulating cell cycle