[Comparison of the effect of Angelica polysaccharide, platelet-derived growth factor and thrombopoietin on megakaryocytopoiesis].
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
APS, PDGF and TPO have similar effect in stimulating proliferation and inhibiting serum-free-culture induced apoptosis of M-07e cells.
Mo
Shu
S
Department of Hematology/Oncology, Chongqing Children's Hospital, Chongqing Medical University, Chongqing 400014, China.
Yu
Jie
J
Yang
Mo
M
Li
Zhi-guang
ZG
Li
Gui-xia
GX
Zhang
Lei
L
D029969:Angelica / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles / Q000494:pharmacology*; D002232:Carbocyanines / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D008533:Megakaryocytes / Q000187:drug effects* / Q000502:physiology; D009930:Organic Chemicals / Q000494:pharmacology*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D034061:Thrombopoiesis; D013926:Thrombopoietin / Q000494:pharmacology*
D001562:Benzimidazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D010982:Platelet-Derived Growth Factor; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; C030750:polysam 1; D013926:Thrombopoietin; D053148:Caspase 3