Fang Gui-Lun, Gao Rui-Lan, Lin Xiao-Jie, Jin Jin-Mei
Institite of Hematology, Affiliated Hospital, Zhejiang University of Traditional Chinese Medicine, Hangzhou 310006, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2007 Aug;15(4):776-9.
The study was purposed to investigate the effects of the panaxadiol saponin (PDS) from Ginseng on proliferation and differentiation of human CD34(+) cells from human bone marrow. Highly purified CD34(+) cells were isolated from human bone marrow by using the Dynal CD34 Cell Selection System (Dynal, Norway). The cells were exposed to PDS at various concentrations in both agar semi-solid culture of CFU-Mix and suspension culture of myeloid and erythroid cells in order to observe the effects of PDS on proliferation of CD34(+) cells. The cells were marked with 4 kinds of monoclonal antibody in related with their differentiation toward to myeloid and erythroid lineages, then examined by flow cytometry (FACS) after being incubated with PDS for 14 days. The results showed that the number of CD34(+) cells was 1.0 +/- 0.15% out of marrow nuclear cells after being purified by Dynal beads system. The enrichment of CD34(+) cells reached to 86.8 +/- 2.8%. The best efficiency in promoting proliferation of CD34(+) cells in vitro was obtained when the concentration of PDS was 25 mg/L, the formation of CFU-Mix colonies significantly increased by 56.3 +/- 3.5% over those of no-PDS control (p < 0.01). The results from suspension culture revealed that myeloid cells elevated in a dose-dependent manner with a peak increasing rate of 35.6 +/- 3.2%, and erythroid cells significantly increased by 22.3 +/- 2.1% over those of no-PDS control (all p < 0.01). After incubation with PDS for 14 days, number of CD33(+) cells increased in a dose-dependent manner with a peak increasing rate at 50 mg/L. CD71(+) cells reaching the peak were at 25 mg/L, while G-A(+) cells were increased by 7.2 +/- 1.3% (p < 0.01) at 10 mg/L, but the number of CD15(+) cells was not found to be changed before and after treating with PDS. It is concluded that PDS not only enhance the proliferation of CD34(+) cells, but also induce differentiation of CD34(+) cells toward to myeloid and erythroid lineages. PDS may play the roles as like hematopoietic growth factor, or provide synergistic effects on growth factor in the regulation of hematopoiesis.
本研究旨在探讨人参二醇皂苷(PDS)对人骨髓CD34(+)细胞增殖和分化的影响。采用Dynal CD34细胞分选系统(挪威Dynal公司)从人骨髓中分离出高度纯化的CD34(+)细胞。将细胞置于不同浓度的PDS中,分别进行CFU-Mix琼脂半固体培养以及髓系和红系细胞的悬浮培养,以观察PDS对CD34(+)细胞增殖的影响。用4种与髓系和红系分化相关的单克隆抗体标记细胞,与PDS孵育14天后,通过流式细胞术(FACS)检测。结果显示,经Dynal磁珠系统纯化后,CD34(+)细胞占骨髓有核细胞的比例为1.0±0.15%,CD34(+)细胞的富集率达到86.8±2.8%。当PDS浓度为25 mg/L时,体外促进CD34(+)细胞增殖的效果最佳,CFU-Mix集落形成比无PDS对照组显著增加56.3±3.5%(p<0.01)。悬浮培养结果显示,髓系细胞呈剂量依赖性升高,峰值增加率为35.6±3.2%,红系细胞比无PDS对照组显著增加22.3±2.1%(均p<0.01)。与PDS孵育14天后,CD33(+)细胞数量呈剂量依赖性增加,在50 mg/L时达到峰值增加率。CD71(+)细胞在25 mg/L时达到峰值,而G-A(+)细胞在10 mg/L时增加了7.2±1.3%(p<0.01),但CD15(+)细胞在用PDS处理前后数量未发现变化。结论是,PDS不仅能增强CD34(+)细胞的增殖,还能诱导CD34(+)细胞向髓系和红系分化。PDS可能起到造血生长因子的作用,或在造血调控中对生长因子提供协同效应。