Abstract
Although the compatibility of Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) has favorable effect on promoting hematopoiesis in traditional Chinese medicine (TCM), the main active components and pharmacological mechanism are unknown. We investigated the five active components and its mechanisms in vitro and in vivo. Five active components of Astragalus glycosides (AST), Formononetin (FRM), Ferulic acid (FRA), Calycosin (CAL), and Calycosin-7-glucoside (CLG), which could be absorbed in intestinal tract, were detected in this study. The peripheral blood, hematopoietic growth factors (HGFs), and hematopoietic progenitor cells (HPCs) colony were observed to evaluate the effect of these five active components promoting hematopoiesis. Furthermore, hematopoietic stem cell (HSC) proliferation, aging, cycle, and related proteins were detected to explore the mechanism of these five components promoting HSC proliferation. i) The in vivo experiments showed that the combination of the five active components could remarkably increase the number of RBCs, WBCs, PLTs, and content of Hb in peripheral blood and the area of bone marrow hematopoietic tissue, as well as thrombopoietin (TPO), erythropoietin (EPO), granulocyte-macrophage colony stimulating factor (GM-CSF), and colony of CFU-GM, CFU-MK, CFU-E, and BFU-E in serum. Each of these five components promoted the recovery of RBCs and Hb, and increased TPO, CFU-MK, and CFU-E. All components except for AST increased the CFU-GM. FRA increased the number of WBCs, the area of bone marrow hematopoietic tissue, and BFU-E. FRA and AST promoted PLT recovery. FRA and CAL improved the content of GM-CSF. FRA, CAL, and CLG improved the content of EPO. ii) The in vitro experiments showed that FRA, FRM, and AST significantly promoted cell proliferation, reduced the positive rate and G0/G1 cells, and increased G2/M + S cells and the expression of cyclin D1 and CDK4 proteins in aging HSCs. Furthermore, the combination of five components had the best effect. Taken together, the five active components of AST, FRM, FRA, CAL, and CLG were the main pharmacodynamic substances of the AR-ASR compatibility, which promoted hematopoiesis. The combination of them had a synergistic effect. The mechanism of promoting hematopoiesis may be relevant to regulating cyclin-related proteins, promoting cell cycle transformation, and promoting HSC proliferation.
Highlights
The compatibility of ASR compatibility (AR) with ASR is commonly used to cure disease in traditional Chinese medicine
In comparison with the BLK group, blood assay showed a significant decrease in white blood cells (WBCs), PLT, red blood cells (RBCs), and Hb (Figures 4a-d), and the area of bone marrow hematopoietic tissue diminished in the model group (Figures 4e, f)
We explored the effect of all drugs on HGFs, and the results showed that compared with the model, the GM-CSF level was remarkably increased in some drug-treated groups (FRA, CAL, active components combination group (ACC), and AA; Figure 5A), the EPO level was dramatically increased in some drug-treated groups (FRA, CAL, CLG, ACC, and AA; Figure 5B), and the TPO level was dramatically increased in all drug-treated groups (Figure 5C)
Summary
The compatibility of AR with ASR is commonly used to cure disease in traditional Chinese medicine. DBT can promote erythropoiesis, and its main active components include astragalus saponin III, AST, CAL, FRM, and FRA (Zhang et al, 2012). The compatibility of AR and ASR (1:1) had the strongest effect (Zhang et al, 2017) and could promote active components [FRA, FRM, AST, CAL, and CLG (calycosin7-glucoside)] dissolution and absorption in isolated small intestine in rats (Li et al, 2017). It is suggested that the active components (FRA, FRM, AST, CAL, and CLG) may be the main effective substances of the compatibility of AR and ASR that promote hematopoiesis. In order to reveal the main pharmacodynamic substances and characteristics of the compatibility of AR-ASR on hematopoiesis, first, we studied the promoting hematopoietic action of five main components and their compatibility in bone marrow hematopoietic inhibition mice. Based on the compatibility of five main active components of AR-ASR promoting hematopoietic effect, the aging model of HSCs was further adopted to study the effect of five components and their compatibility on promoting the proliferation of aging HSCs
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