Abstract

A herbal decoction (Danggui Buxue Tong, DBT) is a traditional Chinese medicine formula comprising Danggui (Radix Angelicae) and Huang Qi (Radix Astragali), which has been used for promotion of “blood production” for centuries and have shown favorable effects on thrombocytopenia. Previously, we have showed that it has significant effects on promoting hematopoiesis and thrombopoiesis (Yang M et al. J Ethnopharmacol, 2009). Many chemical components have been identified in DBT. Among them is the polysaccharides fraction of Angelica sinensis and Radix Astragali. Our study has demonstrated that Angelica Polysaccharide (APS) promotes hematopoiesis and platelet recovery in a mouse model (Liu et al, BMC Complement Altern Med, 2010). In this present study, we further assess the effect of Astragalus Polysaccharide (ASPS) on hematopoiesis and thrombopoiesis in-vivo and in vitro. A myelosuppression mouse model (4-Gy-irradiated) was treated with ASPS (10 mg/kg/day) and TPO (1 mg/kg/day). Peripheral blood cells from ASPS, TPO and vehicle-treated samples were counted on days 0, 7, 14 and 21. Using the colony-forming unit (CFU) assays, we determine the effects of ASPS on the hematopoietic stem/progenitor cells and megakaryocytic lineages. Analyses of Annexin V, Caspase-3, and Mitochondrial Membrane Potential were also conducted in HL-60 cells and megakaryocytic cell line M-07e. The effects of ASPS on cells treated with Ly294002, a PI3K inhibitor and the effect of ASPS on the phosphorylation of AKT were further studied. Results showed that ASPS, like TPO, significantly enhanced the recovery of WBC and platelet count, and bone marrow CFU-GM and CFU-MK formation (n=8) in this model. Morphological examination of bone marrows also showed that ASPS treatment significantly increased the recovery of hematopoietic stem/progenitor cells and megakaryocytic series. We further analyzed the in vitro effect of ASPS on CFU-MK and CFU-GM formation. ASPS (50-100 ug/ml) enhanced TPO (50 ng/ml) -induced CFU-MK (p=0.05, n=4), and CFU-GM formation (p=0.04, n=4). However, ASPS alone (1-500 ug/ml) did not show significant effect on CFU-MK and CFU-GM proliferation (n=4). Moreover, the anti-apoptotic effect of ASPS in HL-60 and M-07e cells were also demonstrated by using Annexin-V, Caspase-3, and JC-1 assays. Addition of Ly294002 alone increased the percentage of cells undergoing apoptosis. However, additional of ASPS to Ly294002-treated cells reversed the percentage of cells undergoing apoptosis. Furthermore, addition of ASPS significantly increased the phosphorylation of AKT. Here, we showed that ASPS, the polysaccharide from the root of Radix Astragali, has hematopoietic and thrombopoietic activities in a mouse model. This effect is likely to result from the stimulation of cell proliferation through the activation of PI3K/AKT pathway, the activation of which prevents cells from undergoing apoptosis.

Disclosures:

No relevant conflicts of interest to declare.

Author notes

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