摘要
目的:探索肿节风总黄酮对骨髓基质细胞-巨核细胞共培养体系中巨核细胞分化成熟,以及基质细胞状态的影响。方法:模拟骨髓微环境,建立KM小鼠骨髓基质细胞和巨核细胞共培养体系,实验分为空白对照组、阿糖胞苷模型组、二甲基亚砜(DMSO)对照组、肿节风总黄酮组7.80、3.90、1.95μg/ml剂量组,每组3个复孔。各组培养4天后分离巨核细胞进行涂片、染色、统计多倍体数目,流式细胞术分析巨核细胞CD61表达量,消化培养体系中贴壁的基质细胞,流式细胞术检测其凋亡情况,ELISA检测培养体系中TPO和SDF-1的含量。结果:与阿糖胞苷模型组相比,肿节风总黄酮7.80μg/ml和3.90μg/ml剂量组中多倍体巨核细胞数目显著增加,肿节风总黄酮7.80μg/ml剂量组巨核细胞CD61表达水平明显提高。肿节风总黄酮7.80μg/ml和3.90μg/ml剂量组显著降低基质细胞的凋亡率。肿节风总黄酮7.80、3.90、1.95μg/ml剂量组显著提高培养体系中TPO和SDF-1的含量。结论:肿节风总黄酮可能是通过抑制共培养体系中基质细胞凋亡促进"量"的增加,增进其分泌细胞因子TPO、SDF-1影响"质"的变化,改善骨髓微环境,促进巨核细胞分化成熟和产生更多的血小板。
Objective： To investigate the effect of flavonoids sarcandrae on megakaryocyte differentiation and stromal cell status in co-culture system of bone marrow stromal cells and megakaryocytes. Methods： A co-culture system of bone marrow stromal cells and megakaryocytes of KM mice was established to simulate the bone marrow microenvironment. The experiment was divided into the control group,Ara-C model group,DMSO control group,7. 80 μg/ml,3. 90 μg/ml and 1. 95 μg/ml Flavonoids Sarcandrae groups,3 holes in each group. After 4 days＇ culture,megakaryocytes of co-culture system were separated and stained,and the number of polyploid was counted. The expression of CD61 in megakaryocytes was evaluated by flow cytometry. The apoptosis of bone marrow stromal cells in co culture system was detected by flow cytometry. Contents of TPO and SDF-1 in co culture system were detected by ELISA. Results： Compared with Ara-C group,the number of polyploid megakaryocytes significantly increased at doses of 7. 80 and 3. 90 μg/ml Flavonoids Sarcandrae（ P〈0. 01）,7. 80 μg/ml Flavonoids Sarcandrae significantly increased the expression of CD61 in megakaryocytes（ P〈0. 01）. 7. 80 and 3. 90 μg/ml Flavonoids Sarcandrae significantly reduced the apoptosis rate of stromal cells in co-culture system（ P〈0. 01 and P〈0. 05）. Levels of TPO and SDF-1 in the culture system were significantly increased by 7. 80,3. 90 and 1. 95 μg/ml Flavonoids Sarcandrae（ P〈0. 01）. Conclusion： Flavonoids Sarcandrae promotes the increase of ＂ quantity＂ by the inhibition of stromal cell apoptosis,and promotes the differentiation and maturation of megakaryocytes by promoting the secretion of cytokines TPO and SDF-1,which influences the change of ＂ quality＂. These could improve the bone marrow microenvironment and promote the differentiation and maturation of megakaryocytes to produce platelets.
出处
《中药药理与临床》
CSCD
北大核心
2017年第6期72-76,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金(81260653)
江西省自然科学基金资助项目(20151BAB205078)
关键词
肿节风总黄酮
骨髓基质细胞
巨核细胞
共培养体系
flavonoids sarcandrae（肿节风总黄酮）
bone marrow stromal cell
megakaryocyte
co-culture system
