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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 investigate the effect and mechanism of Angelica polysaccharide (ASP) on the hematopoietic function of bone marrow in the aging model rats. Methods Forty male SD rats aged 6 to 8 weeks were randomly divided into normal group (n = 10), normal group (n = 10), aging model group (n = 10) . On the second day after the injection of the drug, peripheral blood samples were taken from the eyeball blood to measure the total number of bone marrow mononuclear cells (BMNCs) per femur. CCK8 was used to measure the proliferation of BMNCs. Flow cytometry was used to detect the relationship between proliferative cycle and reactive oxygen species (ROS) ); The colonies of BMNCs were detected by CFU-Mix culture. The percentage of senescent BMNCs was observed by aging β-galactosidase (SA-β-Gal) staining. The total antioxidant capacity -AOC); P53 and P21 protein expression was detected by Western blotting; P53 protein expression and localization were detected by laser scanning confocal microscope. Results Compared with aging model group, the total number of peripheral blood erythrocytes, platelets and white blood cells in ASP model group was decreased; the number of femur BMNCs was increased; the proliferation ability was increased; the CFU-Mix number was increased; the percentage of SA-β-Gal positive BMNCs Significantly decreased; the proportion of cells in G1 phase decreased, the proportion of S phase cells increased, the intracellular ROS content was significantly reduced; T-AOC increased; P53 and P21 were significantly up-regulated. Conclusion ASP can delay or antagonize D-galactose-induced senescence in rat bone marrow hematopoietic cells. The mechanism may be related to the inhibition of oxidative stress by ASP and the down-regulation of p53 / p21 pathway.