Effects Of Angelica Polysaccharides On Adhesion Molecules Expression And Adhesive Function In Murine Bone Marrow Stromal Cell

Bone marrow stromal cell (BMSC) is an important component of bone marrow microenvironment.It can secrete various cytokines and adhere to hematopoietic cells directly to regulate their proliferation,differentiation mobilization and homing. BMSC's superficial adhesion molecule involve in direct contactation between BMSC and hematopoietic cell.It is important that their integrality of construction and function protect haematopoietic stability.Adhesion molecule conclude integrin family,selectin family, immunoglobulin gene super family and so on.Intercellular adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are the main adhesion molecules of immunoglobulin gene super family.They mediate adhesion between BMSC and hematopoietic cell and assist hematopoietic cell to locate microenvironment.They can also involve in forming multimer of hematopoietic cell and stromal cell and mobilizing hematopoietic stem progenitor cells (HSPC) to peripheral blood. Recently,adhesion molecules of ICAM-1 and VCAM-1are hot research spots.There are thousands of years that Angelica is an important Chinese crude drug to nourish blood and promote blood flow.Angelica polysaccharides(APS) is one of the main chemical composition in it.The past researches discovered that APS could obviously promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-MK in health and anaemic mice. APS also could promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-Mix in the human being.APS could mobilize HSPC from bone marrow into peripheral blood. And APS in combination with rhG-CSF may cooperate with each other. HSPC mobilized by APS can reconstruct long-term hematopoiesis in hematopoiesis failure mice. Now there are many reports about the effect of BMSC in mobilization of HSPC,but now there is no report about the effect of APS on the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro. This study provide a new idea for improving bone marrow microenvironment and promoting mobilization of HSPC.In this research experimental animal model is BALB/c mice.There are two parts in this research:1. Effect of different cell culture density and serums on growth of mice BMSC in vitro. 2. Effect of APS on cell proliferation,the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro.Our research mainly study that APS improve hematopoietic microenvironment and participate in mobilizable mechanism of HSPC.This findings will provide theoretical and experimental evidence for exploiting and utilizing Angelica.1. Study on the effect of different cell culture density and serums on growth of mice BMSC in vitroObjective: To study the effect of different cell culture density and serums on growth of mice BMSC in vitro.Methods: BMSC were cultured in DMEM/F12 containing different cell density and serums. The rate of adhered cells, cell viability by trypan blue test,proliferation of BMSC by MTT and cell morphological structure under light microscope were observed.Results: The cultured BMSC proliferated well in DMEM/F12 containing 10%~20% serums and the density of (1~4)×106/mL.It proliferated better in horse serum than fetal bovine serums (P<0.05).Its types were also different in bovine serum,fetal bovine serums and horse serum (P<0.05).Conclusion: There is influence on the growth of mice BMSC cultured in different cell density and serums.2. Study on effects of APS on adhesion molecules expression and adhesive function in murine BMSCObjective: To explore the effect of APS on BMSC's proliferation,the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule -1 (VCAM-1) in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC.Methods: Observe the effect of APS in digestion time of the murine BMSC under inverted microscope,cell proliferation and the binding capacity of murine bone marrow mononuclear cell to BMSC by MTT assay and the expressions of ICAM-1 and VCAM-l of murine BMSC by flow cytometry.Results: 1) With 100, 200 mg/L APS, the digestion time of murine BMSC has obviously become shorter compared with control group. 2) The cultured cells proliferated well in DMEM/F12 containing 50 mg/L and 100 mg/L APS. 3) APS of 200 mg/L and 300 mg/L decreased the expression of ICAM-l, APS of 50,100,200 and 300 mg/L APS decreased the expression of VCAM-1.The expression of ICAM-l and VCAM-1 also decreased by intraperitoneal injection of APS.4) With 100,200 and 300 mg/L of APS, the adhesive rate was inferior than control group.Conclusion: APS can promote BMSC's proliferation, but it decreases the digestion time of murine BMSC,the expression of ICAM-l and VCAM-1 and the adhesive rate of murine bone marrow mononuclear cell to BMSC.To sum up, APS can promote BMSC's proliferation and decreases the expression of ICAM-l and VCAM-1. It will provide experimental evidence for exploiting and utilizing Angelica that APS is further researched in improving bone marrow microenvironment and promoting mobilization of HSPC.