Xiao Bin, Xu Yue, He Han, Jiang Qian-Li, Li Su-Yi, Shu Hui-Ying, Liang En-Yu, Yi Zheng-Shan, Ye Jie-Yu, Huang Lin-Fang, Liu Chang, Meng Fan-Yi, Yang Mo
Institute of Hematology, Jinan University, Guangzhou 510632, Guangdong Province, China; Department of Hematology, Nanfang Hospital, Southern Medical Uniersity, Guangzhou 510515, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Oct;21(5):1243-7. doi: 10.7534/j.issn.1009-2137.2013.05.032.
This study was aimed to assess the effect of Astragalus Polysaccharide (ASPS) on in-vitro hematopoiesis. CFU-GM assays were used to determine the effect of ASPS and thrombopoietin (TPO) on granulocytic-monocyte progenitor cells. The CFU assays were also used to investigate the effect of ASPS on the proliferation of HL-60 cells.HL-60 cells were cultured with serum-free RPMI 1640 medium and treated with or without of different concentrations of ASPS. After 72 h incubation, the number of cells were counted.In addition, the caspase-3 and JC-1 expression was determined by flow cytometry with Annexin V/PI double staining. The results showed that ASPS (100, 200 µg/ml) and TPO (100 ng/ml) significantly promoted CFU-GM formation in vitro. Various concentrations of ASPS and TPO also promoted the colony formation of HL-60 cells, the largest effect of ASPS was observed at a concentration of 100 µg/ml. There were no synergistic effects between TPO and ASPS on cellular proliferation. The results also showed that ASPS significantly protected HL-60 cells from apoptosis in condition of serum-free medium culture, suppressed caspase 3 activation, and reduced the cell apoptosis. It is concluded that ASPS can significantly promote the formation of bone marrow CFU-GM and the proliferation of HL-60 cells, the optimal concentration of ASPS is at 100 µg/ml. In the absence of serum inducing apoptosis, ASPS also significantly reduced the apoptosis of HL-60 cells via suppressing the activation of caspase-3.
本研究旨在评估黄芪多糖（ASPS）对体外造血的影响。采用CFU-GM检测法来确定ASPS和血小板生成素（TPO）对粒系-单核系祖细胞的作用。CFU检测法还用于研究ASPS对HL-60细胞增殖的影响。HL-60细胞用无血清RPMI 1640培养基培养，并分别用不同浓度的ASPS处理或不处理。孵育72小时后，对细胞进行计数。此外，通过Annexin V/PI双染流式细胞术检测caspase-3和JC-1的表达。结果显示，ASPS（100、200μg/ml）和TPO（100 ng/ml）在体外显著促进CFU-GM的形成。不同浓度的ASPS和TPO也促进了HL-60细胞的集落形成，在100μg/ml浓度下观察到ASPS的最大作用效果。TPO和ASPS在细胞增殖方面没有协同作用。结果还表明，在无血清培养基培养条件下，ASPS能显著保护HL-60细胞免于凋亡，抑制caspase 3激活，并减少细胞凋亡。结论是，ASPS能显著促进骨髓CFU-GM的形成和HL-60细胞的增殖，ASPS的最佳浓度为100μg/ml。在无血清诱导凋亡的情况下，ASPS还通过抑制caspase-3的激活显著减少HL-60细胞的凋亡。
