当归多糖对小鼠衰老造血干细胞细胞周期蛋白的调控

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (3): 320-324.

当归多糖对小鼠衰老造血干细胞细胞周期蛋白的调控

张先平¹,王乾兴²,陈斌³,刘俊¹,魏强¹,王建伟¹,王亚平¹

1. 重庆医科大学
2. 遵义医学院生物学教研室遵义
3. 山东省枣庄市台儿庄区中医院

收稿日期:2012-11-29 修回日期:2013-01-18 出版日期:2013-03-05 发布日期:2013-03-05
通讯作者: 王亚平 E-mail:ypwangcq@yahoo.cn
基金资助:
当归多糖调控血液干细胞衰老的机理研究;人参总皂苷及单体Rd、Re、Rg1抗放射所致骨髓抑制作用中对HSC自我更新能力的影响

Angelica sinensis polysaccharides regulate aging of mice hematopoietic stem cell through cell cycle protein

Received:2012-11-29 Revised:2013-01-18 Online:2013-03-05 Published:2013-03-05

摘要/Abstract

摘要： 目的观察当归多糖（ASP）对小鼠造血干细胞（HSC）细胞周期调控蛋白表达的影响，探讨ASP调控HSC衰老的可能机制。方法C57BL/6J小鼠随机分为对照组、衰老组、ASP干预对照组和ASP干预衰老组，衰老组采用X线全身均匀照射，建立小鼠HSC衰老模型；ASP干预衰老组在照射期间给予ASP灌胃；对照组和ASP干预对照组分别给予NS和ASP灌胃。免疫磁珠分离HSC，β-半乳糖苷酶（SA-β-Gal）染色和混合集落培养（CFU-Mix）观察HSC生物学特性变化；流式细胞术分析细胞周期；Western blot检测 P16、P21、CDK2、CDK6、CyclinD及CyclinE表达。结果与对照组比较，X线能显著增加衰老对照组HSC SA-β-Gal染色阳性率、G1期比例及P16、P21表达；降低CFU-Mix、S期比例及CDK6、CyclinD和CyclinE表达表达。与衰老组比较，ASP能显著抑制衰老HSC SA-β-Gal染色阳性率、G1期比例及P16和P21表达的增加；抑制S期比例、CFU-Mix、CDK6、CyclinD及CyclinE表达的减少；而对CDK2表达无影响。结论 ASP可能通过调节P16、P21、CDK6、CyclinD及CyclinE表达延缓小鼠HSC衰老。

关键词: 当归多糖, 细胞衰老, 造血干细胞, 细胞周期

Abstract: Objective The effect of angelica sinensis polysaccharides (ASP) on the expression of contol cell cycle protein in mice hematopoietic stem cells (HSCs) were observed to explore the underlying mechanism that ASP delay aging of HSCs in vivo. Methods C57BL/6J mice were randomly divided into control group, ASP regulate control group, aging group, ASP regulation aging group. Mice were uniformly explored in X-ray to erect model of aging. ASP regulation aging groups mice were treated with ASP by intragastric administration during X-ray irradiation. While control and ASP regulation control groups were treated with equal-volume NS and ASP by intragastric administration. Mouse HSCs were isolated by magnetic cell sorting and cultured in vitro. Senescence-associated β-galactosidase (SA-β-Gal) staining was used to detect aging HSCs. Cell cycles analysis and CFU-Mix cultivation were used to evaluate the capability of colony forming in HSCs. Cell cycles were detected by flow cytometry. The expressions of p16, p21,CDK2,CDK6, cyclinD and cyclinE were determined by western blot analysis. Results Exogenous X-ray irradiation induced HSCs aging was compared with control group. Biological feature of HSCs in aging group as follows: The percentage of SA-β-Gal positive cells, the ratio of G1 stages and the expression of p16 and p21 protein were significantly increased , the expression of CDK6、CyclinD、CyclinE, the ratio of S stages and the capacility of colony forming in HSCs were decreased. ASP could significantly decrease the number of SA-β-Gal positive cells, the ratio of G1 stages and downregulate the expression of p16 and p21 protein in HSCs contrast to aging control group. In addition, ASP could remarkably increase the ratio of S stages, the capacility of colony forming and upregulate the expression of CDK6、CyclinD、CyclinE in HSCs compared with aging group. There was no significant difference in the protein expression of CDK2 in HSC. Conclusions ASP could delay senescence HSCs aging which maybe partly ascribed to the regulation of o cell cycle control gene.

Key words: Angelica sinensis polysaccharides, Hematopoietic stem cells, Cell aging, Cell cycle

中图分类号:

张先平王乾兴陈斌刘俊魏强王建伟王亚平. 当归多糖对小鼠衰老造血干细胞细胞周期蛋白的调控[J]. 基础医学与临床, 2013, 33(3): 320-324.

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