Effect Of Astragaloside Ⅳ On The Proliferation And Cell Factor Expression In HBMSC

Objective：Astragaloside IV is the monomer composition of Astragalus membranaceus,that is tonifying qi of traditional Chinese medicine. This experiment was to investigate theeffect of Astragaloside IV on the proliferation and the expression of cell factor in human bonemarrow mesenchymal stem cells(hBMSCs) and to confirm the mechanism.Methods：The isolation,purification and training of hBMSCs from bone marrow bypercoll isodensity centrifugation and adherent method. To observe the growth andmorphological changes of the primary culture and subculture hBMSCs by invertedmicroscope. The CD44, CD71, CD45, CD34surface marker of hBMSCs were detected byflow cytometer. To determinate the proliferation index(OD value) of hBMSCs with differentconcentration (0,20,40,80,160,320mg/mL) Astragaloside IV for72hours were evaluated byMTT, choosing the third generation hBMSCs in logarithmic phase. To select the best ofpromoted concentration as a reference for follow-up test the concentration of drugintervention. The next tests select concentration with160mg/mL of Astragaloside IVaccording to experimental result earlier stage. The third generation hBMSCs in logarithmicphase is divided into control group and160mg/mL drug group. The expressions of SCF,VEGF, SDF-1, GM-CSF mRNA were measured by Real-time PCR, after the hBMSCs wereevaluated with160mg/mL Astragaloside IV for72hours.Results：The cells grew by static adherence, cell morphology was spindle shape andranged like net or swirls. The hBMSCs were isolated successfully. The surface marker CD44,CD71of mesenchymal stem cells were high expression,.and the surface marker CD34, CD45of Hematopoietic stem cells were negative. Astragaloside IV at20,40,80,160,320mg/mLsignificantly increased the proliferation of hBMSCs (P<0.05), especially the group of160mg/mL had the most obvious effect. After the drug reaching a certain concentration, humanbone marrow mesenchymal stem cells no longer increases with the drug concentration.Astragaloside IV could stimulated the expressions of SCF, VEGF, SDF-1mRNA, butGM-CSF mRNA were not change significantly.Conclusions： Astragaloside IV could increase the proliferation of hBMSCs,Astragaloside IV could improve the expressions of the SCF, VEGF, SDF-1mRNA,that relate to proliferation mechanism.