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纸质出版: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:
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
0
/G
1
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
4
,低于对照组[(9.8±2.1)/10
4
,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。
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
4
vs (9.8±2.1)/10
4
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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