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当归多糖对衰老模型大鼠骨髓造血功能的影响及其机制
张岩岩 李静 贾道勇 张梦思 徐春燕 景鹏伟 李成鹏 王璐 王顺和 王亚平*
解剖学报 ›› 2014, Vol. 45 ›› Issue (6) : 785-792.
当归多糖对衰老模型大鼠骨髓造血功能的影响及其机制
Effect of angelica polysaccharide on hematopiesis in aging rats and its biological mechanism
目的 探讨当归多糖（ASP）对衰老模型大鼠骨髓造血功能的影响及其机制。方法 6~8周雄性SD大鼠40只，随机分为正常组（n=10）、ASP正常组（n=10）、衰老模型组（n=10）和ASP衰老模型组（n=10）。药物注射完成第2天，取眼球血检测外周血常规，取股骨测定每根股骨骨髓单个核细胞（BMNCs）总数；CCK8测定BMNCs增殖能力；流式细胞术检测BMNCs增殖周期与活性氧簇（ROS）含量；造血祖细胞混合集落（CFU-Mix）培养检测BMNCs形成集落能力；衰老β半乳糖苷酶（SA-β-Gal）染色观察衰老BMNCs百分率；酶学法检测细胞总抗氧化能力（T-AOC）；Western blotting检测P53和P21蛋白表达；激光扫描共焦显微镜检测P53蛋白表达及定位。 结果 与衰老模型组比较，ASP衰老模型组外周血红细胞、血小板和白细胞总数下降得到抑制；股骨BMNCs细胞数升高；增殖能力提高；形成CFU-Mix数增加；SA-β-Gal染色阳性BMNCs百分率显著降低； G1期细胞比例降低，S期细胞比例升高，细胞内ROS含量显著降低； T-AOC升高: P53和P21表达显著上调。 结论 ASP能延缓或拮抗D-半乳糖致大鼠骨髓造血细胞衰老，其机制可能与ASP抑制氧化应激，下调p53/p21通路有关。
Objective To explore the effect and mechanism of angelica sinensis polysaccharide（ASP）on hematopiesis in aging rats and to provide theoretical and experimental evidences for effective natural medicine of antiaging or protecting hematopoietic function. Methods Male SD rats（n=40）aging from 6 to 8 weeks old were randomly divided into normal group(NG, n=10)，ASP normal group(ANG, n=10), aging model group(AMG, n=10) and ASP aging model group(AAMG, n=10). After 2 days of the treatment, the eyeball blood was collected for the peripheral blood routine test; The femur was taken to count the number of each femur bone marrow mononuclear cells (BMNCs). The proliferation of BMNCs was detected by CCK-8. The distribution of cell cycles and reactive oxygen species (ROS) levels were analyzed by flow cytometry(FCM). The capability of colony formation was examined by CFU-Mix cultivation. The ratio of the SA-β-Gal staining positive BMNCs was counted. The total antioxidant capacity of the cell was detected by enzymatic assay. The aging related proteins of P53 and P21 were detected by Western blotting. Results Compared with the aging model group, in the ASP aging model group,the decrease in the amounts of the peripheral blood RBC, PLT and WBC was evidently inhibited; the proliferation of the BMNCs was enhanced;the colony formation of CFU-Mix was markedly increased; the percentage of SA-β-Gal, the ratio of G1 stages and the product of ROS positive cells were significantly reduced; the ratio of S stages was markedly promoted; the total antioxidant capacity of the cell was significantly promoted; the expression of the aging related protein of P53 and P21 was evidently down-regulated.
Conclusion ASP can protect hematopoietic function from decline by antagonize D-galactose that induced oxidative stress, suggesting the mechanism of ASP protecting hematopiesis may regulate p53/p21 pathway.
当归多糖 / 衰老模型 / 骨髓单个核细胞 / 免疫印迹法 / 大鼠
Angelica polysaccharide / Aging model / Bone marrow mononuclear cell / Western blotting / Rat
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当归多糖调控血液干细胞衰老的机理研究
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