Mo Shu, Yu Jie, Yang Mo, Li Zhi-guang, Li Gui-xia, Zhang Lei
Department of Hematology/Oncology, Chongqing Children's Hospital, Chongqing Medical University, Chongqing 400014, China.
Zhonghua Er Ke Za Zhi. 2008 Jan;46(1):45-8.
To investigate the effect of Angelica polysaccharide (APS), platelet-derived growth factor (PDGF) and thrombopoietin (TPO) on the proliferation and apoptosis of human megakaryocytic cell line M-07e.
Cell count and the viability testing of M-07e cells (trypan blue exclusion assay) were performed at 24 hours, 48 hours and 72 hours after treatment with APS, PDGF or TPO. Three apoptosis related flow cytometric assays including Annexin V, Caspase-3 and JC-1 were performed to determine apoptotic rate of each group at 72 hours after the treatment.
After the incubation, the number of M-07e cells in the APS, PDGF and TPO group increased and the viabilities of the three groups were significantly higher than the control group (P < 0.05). The dead cells in the APS, PDGF and TPO group were (19.41 +/- 7.59)%, (21.38 +/- 7.25)% and (18.77 +/- 8.00)%, respectively by flow cytometry using Annexin V method, which were significantly lower compared to the control group (34.33 +/- 5.46)%. The expression of the activated caspase-3 in the group of APS, PDGF and TPO were (12.27 +/- 5.18)%, (12.39 +/- 6.26)% and (13.75 +/- 8.25)%, the APS and PDGF group decreased significantly compared to the control group (18.92 +/- 6.09)%. The ratio of total cell deaths in the APS, PDGF and TPO group were (23.64 +/- 6.69)%, (28.00 +/- 10.05)% and (27.99 +/- 8.99)%, the ratio in APS group decreased significantly compared to the control group (39.48 +/- 11.86)% by JC-1 method. Differences between APS and PDGF groups and between APS and TPO groups were not statistically significant.
APS, PDGF and TPO have similar effect in stimulating proliferation and inhibiting serum-free-culture induced apoptosis of M-07e cells.
探讨当归多糖（APS）、血小板衍生生长因子（PDGF）和血小板生成素（TPO）对人巨核细胞系M-07e增殖和凋亡的影响。
用APS、PDGF或TPO处理M-07e细胞24小时、48小时和72小时后，进行细胞计数和细胞活力检测（台盼蓝排斥试验）。在处理72小时后，进行三项与凋亡相关的流式细胞术检测，包括膜联蛋白V、半胱天冬酶-3和JC-1检测，以确定每组的凋亡率。
孵育后，APS、PDGF和TPO组的M-07e细胞数量增加，三组的细胞活力均显著高于对照组（P<0.05）。采用膜联蛋白V法通过流式细胞术检测，APS、PDGF和TPO组的死细胞率分别为（19.41±7.59）%、（21.38±7.25）%和（18.77±8.00）%，与对照组（34.33±5.46）%相比显著降低。APS、PDGF和TPO组中活化半胱天冬酶-3的表达分别为（12.27±5.18）%、（12.39±6.26）%和（13.75±8.25）%，APS和PDGF组与对照组（18.92±6.09）%相比显著降低。采用JC-1法检测，APS、PDGF和TPO组的总细胞死亡率分别为（23.64±6.69）%、（28.00±10.05）%和（27.99±8.99）%，APS组与对照组（39.48±11.86）%相比显著降低。APS组与PDGF组之间以及APS组与TPO组之间的差异无统计学意义。
APS、PDGF和TPO在刺激M-07e细胞增殖和抑制无血清培养诱导的凋亡方面具有相似的作用。
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