ABSTRACT
The aim is to study the effect and its mechanism of Astragalus Radix combined with Angelicae Sinensis Radix on the proliferation of hematopoietic stem cells(HSCs) in senescence model. After drug-containing plasma of rats was prepared via intragastric administration, HSCs of mice were cultured in vitro, and then they were divided into blank control group, model group, blank plasma group, Astragalus Radix + Angelicae Sinensis Radix 1∶1 group and 10∶1 group, Angelicae Sinensis Radix plasma group, and Astragalus Radix plasma group. HSCs senescence model was induced by using tert-butyl hydrogen peroxide(t-BHP), and intervened by drug-containing plasma. Cells senescence rate was tested by SA-β-galactosidase staining method; cell cycle distribution was determined by flow cytometry; Cyclin D1, P21, and P53 mRNA were measured with RT-PCR, and Cyclin D1 protein expression was measured by Western blot. Results showed that after being induced by t-BHP, senescence rate of HSCs was increased; cell proliferation ability was decreased; count of G₀/G₁ phase cells was increased; count of G₂/M+S phase cells was reduced; Cyclin D1 expression was down-regulated while P53, P21 expression was up-regulated, which were reversed by Astragalus Radix + Angelicae Sinensis Radix 1∶1 and 10∶1, single Angelicae Sinensis Radix, and single Astragalus Radix plasma. Furthermore, the above effects were most obvious in Astragalus Radix+Angelicae Sinensis Radix 1∶1 group. These results suggested that t-BHP can promote HSCs senescence and reduce cell proliferation ability. Angelicae Sinensis Radix, Astragalus Radix and their combinations can inhibit HSCs senescence, promote HSCs proliferation as well as cell cycle conversion; moreover, the effects of 1∶1 Astragalus Radix+Angelicae Sinensis Radix were strongest. The mechanisms may be related to up-regulating the expression of cell cycle positive regulator, down-regulating the expression of cell cycle negative regulator, thus promoting the cells to enter the proliferation phase from the stationary phase.
ABSTRACT
Objective: To investigate the effects of different proportions of Astragalus and Angelica on the proliferation ability and cell senescence of hematopoietic progenitor cells (HPC) in the mice model of bone marrow hematopoiesis suppression, and to probe the mechanism of Astragalus-Angelica compatibility on promoting hematopoiesis. Methods: ICR male mice were randomly divided into normal group, model group, positive control group of recombinant human granulocyte colony stimulating factor (rhG-CSF), Astragalus group, Angelica group, and different proportion combination groups of Astragalus and Angelica, and the animals in Chinese medicinal herb groups were ig administered, once a day, for 8 d. In the positive control group, rhG-CSF was sc injected on days 6, 7, and 8 of administration. Except for the normal group, the others were received cyclophosphamide (CTX) by ip injection on days 4, 5, and 6 of administration to establish the model of bone marrow hemopoiesis suppression. The mice were killed on day 9 to obtain bone marrow cells for the culture of HPC, and the senescence rate of bone marrow nucleated cell (BMNC) was detected by SA-β-galactosidase staining method. Results: Compared with the model group, Angelica and Astragalus-Angelica with proportions of 5:1, 1:1, and 1:5 could significantly raise colony forming unit-granulocyte (CFU-GM), colony forming unit-megakaryocyte (CFU-MK), colony forming unit-erythroid (CFU-E), and burst forming unit-erythroid (BFU-E) (P < 0.01). Astragalus-Angelica with 10:1 made CFU-MK, CFU-E, BFU-E remarkably increase (P < 0.05, 0.01), and had no effect on CFU-GM. Furthermore, Astragalus-Angelica with 1:1 promoting the formation of HPC was evidently stronger than that of single Astragalus, single Angelica, and other combinations (P < 0.05). Compared with the model group, the senescence positive rate of BMNC was markedly decreased in Astragalus group, Angelica group, and Astragalus-Angelica combination groups (P < 0.01), while being lowest in Astragalus-Angelica with 1:1 proportion (P < 0.05, 0.01). Conclusion: Astragalus, Angelica, Astragalus-Angelica with 10:1, 5:1, 1:1, and 1:5 proportions can prompt the proliferation and differentiation of HPC that bone marrow hematopoiesis is suppressed in mice, inhibit the senescence of bone marrow hematopoietic cell, moreover, the effect of Astragalus-Angelica with 1:1 proportion is the best. It suggests that promoting the proliferation and differentiation of HPC is one of the mechanisms that Astragalus-Angelica prompt hematopoiesis.