[Angelica sinensis polysaccharides delay aging of hematopoietic stem cells through inhibitting oxidative damge].
[Angelica sinensis polysaccharides delay aging of hematopoietic stem cells through inhibitting oxidative damge].
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发表时间：
2013-02
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通讯作者：
Xian-ping Zhang;Qianqian Wang;Bin Chen;Q. Wei;Chun-yan Xu;R. Jiang;Jian-wei Wang;Ya-ping Wang
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作者：
Xian-ping Zhang;Qianqian Wang;Bin Chen;Q. Wei;Chun-yan Xu;R. Jiang;Jian-wei Wang;Ya-ping Wang
目的观察当归多糖（ASP）对小鼠造血干细胞（HSCs）活性氧（ROS）产生、总抗氧化能力（T-AOC）及p16基因表达的影响，探讨ASP延缓HSCs衰老的机制。方法将C57 BL/6 J小鼠随机分为正常组、衰老组和ASP治疗组。用X线（3.0戈伊/8 F）照射小鼠，造成直立老化模型。正常和衰老ASP干预组小鼠灌胃给予ASP，正常和衰老组在X射线照射的同时给予等体积的NS。采用磁性细胞分选法分离小鼠肝星状细胞并进行体外培养。衰老相关的β-半乳糖苷酶（SA-β-Gal）染色用于检测老化的HSC。通过细胞周期分析和CFU-Mix培养评价HSC的自我更新和集落形成能力。流式细胞术检测细胞内ROS的产生，免疫荧光法检测细胞内ROS的产生。采用化学比色法测定总抗氧化能力。实时定量PCR（qRT-PCR）检测p16蛋白的表达。结果外源性X射线照射诱导的HSCs老化与未照射的正常组比较。X射线照射后衰老组HSC生物学特征为：SA-β-Gal阳性细胞百分率、G1期细胞比例、ROS产生量明显增加，p16 mRNA表达上调。HSC集落形成能力和T-AOC降低。ASP能显著降低衰老组HSC中SA-β-Gal阳性细胞百分率、G1期细胞比例和ROS产生量，下调p16 mRNA表达。此外，与未加ASP的衰老组相比，ASP能显著提高HSC的T-AOC和集落形成能力。结论X射线（3.0戈伊/8 F）可诱导小鼠HSC老化。ASP可延缓HSCs的衰老，其机制可能与抑制HSCs氧化损伤和下调p16 mRNA表达有关。
OBJECTIVE The effect of angelica sinensis polysaccharides (ASP) on the production of reactive oxygen specie (ROS), the capability of total anti-oxidant (T-AOC), and the expression of p16 in mRNA level in mice hematopoietic stem cells (HSCs) were observed to explore the underlying mechanism that ASP delay aging of HSCs in vivo. METHOD C57BL/6J mice were randomly divided into normal group, aging group, and the above groups treated with ASP. Mice were uniformly explored in X-ray (3.0 Gy/8 F) to erect model of aging. Normal and aging ASP intervention groups mice were treated with ASP by intragastric administration, while normal and aging groups were treated with equal-volume NS during X-ray irradiation. Mice HSCs were isolated by magnetic cell sorting and cultured in vitro. Senescence-associated beta-galactosidase (SA-beta-Gal) staining was used to detect aging HSCs. Cell cycles analysis and CFU-Mix cultivation were used to evaluate the capability of self-renewing and colony forming in HSCs. The production of ROS in HSCs was evaluated by flow cytometry analysis and immunofluorescence assess, respectively. T-AOC was detected by chemical colorimetric method. The expression of p16 was determined by real-time quantitative PCR (qRT-PCR). RESULT Exogenous X-ray irradiation induced HSCs aging was compared with normal group without irradiation. Biological feature of HSCs in aging group with X-ray irradiation as follows: The percentage of SA-beta-Gal positive cells, the ratio of G1 stages and the production of ROS were significantly increased , the expression of p16 in mRNA level was also upregulated. The capacility of colony forming and T-AOC in HSCs were decreased. ASP could significantly decrease the percentage of SA-beta-Gal positive cells, the ratio of G1 stages and the production of ROS in HSCs, and downregulate the expression of p16 in mRNA level in HSCs contrast to aging group without ASP treatment. In addition, ASP could remarkably increase T-AOC and the capacility of colony forming in HSCs compared with aging group without ASP treatment. CONCLUSION X-ray (3.0 Gy/8 F) could induce mice HSCs aging. ASP could delay senescence HSCs aging which maybe partly ascribed to the inhibition of oxidative damage and the downregulation of p16 mRNA expression.