Zhang Xian-Ping, Wang Qian-Xing, Chen Bin, Wei Qiangi, Xu Chun-Yan, Jiang Rong, Wang Jian-Wei, Wang Ya-Ping
Laboratory of Stem Cell and Tissue Engineering, Department of Histology and Embryology, Chongqing Medical University, Chongqing 400016, China.
Zhongguo Zhong Yao Za Zhi. 2013 Feb;38(3):407-12.
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.
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).
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.
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.
观察当归多糖（ASP）对小鼠造血干细胞（HSCs）中活性氧（ROS）生成、总抗氧化能力（T-AOC）及p16 mRNA水平表达的影响，以探讨ASP在体内延缓HSCs衰老的潜在机制。
将C57BL/6J小鼠随机分为正常组、衰老组及上述两组经ASP处理组。小鼠均接受X射线（3.0 Gy/8 F）照射以建立衰老模型。正常及衰老ASP干预组小鼠通过灌胃给予ASP，而正常组和衰老组在X射线照射期间给予等体积生理盐水。通过磁珠分选法分离小鼠HSCs并进行体外培养。采用衰老相关β-半乳糖苷酶（SA-β-Gal）染色检测衰老的HSCs。利用细胞周期分析和CFU-Mix培养评估HSCs的自我更新能力和集落形成能力。分别通过流式细胞术分析和免疫荧光评估HSCs中ROS的生成。采用化学比色法检测T-AOC。通过实时定量PCR（qRT-PCR）测定p16的表达。
与未照射的正常组相比，外源性X射线照射诱导HSCs衰老。X射线照射衰老组HSCs的生物学特性如下：SA-β-Gal阳性细胞百分比、G1期比例和ROS生成显著增加，p16 mRNA水平表达也上调。HSCs的集落形成能力和T-AOC降低。与未用ASP处理的衰老组相比，ASP可显著降低HSCs中SA-β-Gal阳性细胞百分比、G1期比例和ROS生成，并下调HSCs中p16 mRNA水平的表达。此外，与未用ASP处理的衰老组相比，ASP可显著提高HSCs的T-AOC和集落形成能力。
X射线（3.0 Gy/8 F）可诱导小鼠HSCs衰老。ASP可延缓衰老HSCs的衰老，这可能部分归因于对氧化损伤的抑制和p16 mRNA表达的下调。
