黄芪多糖对粒单核系造血细胞的抗凋亡作用  

Anti-apoptotic Effect of Astragalus Polysaccharide on Myeloid Cells

作　　者：肖彬[1,2] 徐月[1,2] 何涵[2] 江千里[2] 李素毅[2] 舒慧英[3] 梁恩瑜[2] 易正山[2] 叶洁瑜[2] 黄林芳[4] 刘昶[4] 孟凡义[2] 杨默[2] 

机构地区：[1]暨南大学血液病研究所,广东广州510632 [2]南方医科大学南方医院血液科,广东广州510000 [3]重庆医科大学附属儿童医院血液科,重庆400014 [4]中国医学科学院药用植物研究所,北京100193

出　　处：《中国实验血液学杂志》2013年第5期1243-1247,共5页Journal of Experimental Hematology

基　　金：杨默教授的国家自然科学基金(编号为81270580);广东省教育厅人才引进基金;南方医院院长基金(重点);孟凡义教授和杨默教授的广东省教育部省部产学研基金;江千里博士的国家自然科学青年基金(编号为30901367)的支持

摘　　要：本研究探讨黄芪多糖(astragalus polysaccharide,ASPS)体外对骨髓粒单核前体细胞的增殖作用和对HL-60细胞株凋亡的影响及其可能的作用机理。观察不同浓度(0,50,100,200μg/ml)的黄芪多糖和TPO(100 ng/ml)对骨髓集落形成单位CFU-GM和HL-60集落生成的影响。用无胎牛血清(或1%)的改良型RPMI 1640培养液诱导HL-60细胞凋亡,加入或者不加入黄芪多糖共培养72 h后,进行细胞计数并用Annexin V/PI双标记流式细胞术检测凋亡指标Caspase-3和JC-1的表达,并与正常组比较。结果表明,黄芪多糖(100,200μg/ml)与TPO在体外能明显促进人骨髓细胞集落CFU-GM形成,同时黄芪多糖(50,100μg/ml)也能促进HL-60细胞集落生成且其最大作用浓度为100μg/ml,未发现其与TPO具有协同促进增殖的作用。无胎牛血清的RPMI 1640培养液能降低HL-60细胞的增殖,诱导细胞凋亡。HL-60细胞经黄芪多糖作用72 h后,与对照组相比细胞计数明显上升,由19×104个上升到34×104个(P<0.05),caspase-3表达相对于对照组明显下降,分别由10.5%,14.1%降至7.2%(P<0.05),7.8%(P<0.05)。结论:黄芪多糖和TPO均能促进骨髓CFU-GM和HL-60细胞集落的生成;在无胎牛血清RPMI 1640培养液诱导细胞凋亡的情况下,黄芪多糖显著减少HL-60细胞的凋亡,保护HL-60细胞。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 progeni- tor 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 cytometvy with Annexin V/PI double staining. The results showed that ASPS （ 100, 200 Izg/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 tzg/ml. There were no synergistic effects between TPO and ASPS on cellular proliferation. The results also showed that ASPS significantly pro- tected 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 txg/ml. In the absence of serum inducing apop- tosis, ASPS also significantly reduced the apoptosis of HL-60 cells via suppressing the activation of caspase-3.

关 键 词：黄芪多糖 粒单核造血细胞 HL-60 CFU-GM 细胞凋亡 

分 类 号：R979.1[医药卫生—药品]