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Improvation of Ginsenosides Rg1 on Bone Marrow Function and Mechanism in Bone Marrow Depression Mice by Cyclophosphamide

Author: XuShangFu
Tutor: YuLiMei
School: Zunyi Medical College,
Course: Pharmacology
Keywords: ginsenosides Rg1 mouse bone marrow haemopoietic stem cell mesenchyme stem cell endothelium progenitor cell calcium sensitive receptor peripheral blood
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 26
Quote: 2
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Abstract


Objective To study the effects of ginsenosides Rg1(Rg1) on bone marrow(BM) founction in BM depression mice by cyclophosphamide(CY).Calcium-sensing receptor(CaSR) was regarded as a main target,and the potential mechanism of Rg1 was investigated.Methods Male mice were injected CY 150 mg/kg for three days by peritoneal injection(i.p.) for establishing BM depression model.Animals were individed into five groups.They are control,model,high dose Rg1(RH) group and BM depression with low and high dose Rg1(MRL and MRH) groups.Rg1 was administrated according to 7.5 and 15 mg/kg/d for seven days by i.p.The BM cell number was counted by counting plate.The percentage of different cell subpopulation was detected by flow cytometry.The histopathology of of femoral bone sections were examined by H.E.staining.The level of CaSR and mitogen-activated protein kinase (MAPK) mRNA were determined by RT real-time PCR.The protein of CaSR was analyzed by immunofluorescence(IF) staining.The contents of stem cell factor(SCF), stromal cell derived factor 1(SDF-1) and interleukin-3(IL-3) were determined in BM intercellular fluid and serum of peripheral blood(PB) by ELISA methods.Results BM cells were significantly decreased in the model group compared with in the contral group(P＜0.01),but not change in BH group.BM cells were significantly increased in MRH group compared with model group.The percentages of Lin~-Sca-1~+c-kit~+(LSK),Sca-1~-CD44~+CD166~+ and lymphoid system cells(CD3~+) were lower in model than in contral group,as well as CD133~+flk-1~+ in BM.The percentage of MSC of RH group was higher in PB yet.The percentage of CD11b~+ cells was increased PB in model animal compared with contral group.And the percentage CD11b~+ cells were further increased by MRH compared with model group. The contents of SCF and SDF-1 in BM intercellular fluid and SCF,SDF-1 and IL-3 in serum of PB were remarkably outed down in the model group(P＜0.01) compared with contral group.But the contents of SCF,SDF-1 and IL-3 were higher in MRH group than in model group(P＜0.01),as well in PB.In model group,were higher than in contral group.Then,MRL and MRH all decreased the level of CaSR and MAPK mRNA(P＜0.05).Conclusion Rg1 could affect normal BM function at some of degree.RH was not change BM cell number in normal mouse.At the same time,the proportions of CD133~+flk-1~+ in BM and Sca-1~-CD44~+CD166~+ cells in PB,granulocyte and monocyte in PB were increase in normal group.The results hint that Rg1 may increase endothelial progenitor cell(EPC) and mesenchymal stem cell(MSC) in BM and PB of normal mice,respectively.The proportion of LSK,Sca-1~-CD44~+CD166~+ and CD133~+flk-1~+ cells in BM and HSC,MSC and lymphoid system cells in PB all were decreased.These suggested that cell proliferation,differeation and mobilizing of LSK,Sca-1~-CD44~+CD166~+ and CD133~+flk-1~+ cells emerged obvious disfunction in BM with BM depression.The mechanism was related in upregulation of CaSR and MAPK mRNA level and the expression of CaSR protein,and downregultion the contents of SCF and SDF-1 in BM intercellular fluid and SCF,SDF-1 and IL-3 in serum PB.In BM depression state induced by CY,ginsenosides Rg1 could improve BM function,increase BM cell number,and heighten the proportion of LSK, Sca-1~-CD44~+CD166~+ and CD133~+flk-1~+ cells,and improved the morphological changes.Rg1 asle may mobilize BMSC moving to PB.And then,not only the protortions of LSK and Sca-1~-CD44~+CD166~+ cells were significantly increased,but also granulocyte and monocyte.These results suggested that Rg1 may improve the proliferationg,differeation and mobilizing of HSC to PB.The mechanism of Rg1 on recovering BM function involved in downregultion of CaSR and MAPK mRNA level and the expression of CaSR protein,and upregulation the contents of SCF,SDF-1 and IL-3 in BM intercellular fluid and in serum of PB.

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