Effects Of Rg1with Human Cord Blood Stem Cells Transplantation On Hematopoiesis Function Recovery In Radiation Damaged SCID Mice

Objective To study the effects of ginsenosides Rgl (Rgl) with human cord blood stem cells transplantation on hematopoiesis function recovery in radiation damaged SCID mice.Methods The contents of hUCB-MNCs and haemopoietic stem cells were detected by flowcytometry. Severe radiation damage SCID modle was preparaed by sublethal dose X ray administer radiation. The hUCB-MNCs were implantation by vena caudalis of mice. Hemogram of peripheral blood (PB) was determined using auto-omplete blood cytoanalyze. Some of cell subpopulation was detected by flow cytometry. Free intracellular calcium concentration ([Ca2+]i) was detected by fluorescence photometry. The anti-human cell nucleolus antibody-FITC labeled hUCB-MNCs were observed in myeloid tissue field planting and distribution by fluorescence microscope.Results NMCs quantities were (8.62±5.45)×107cells from every cord blood. The content of CD34T cells were0.65-2.48%in hUCB-MNCs. After sublethal dose X ray irradiated using medical linear accelerator, histological section of femoral bone in SCID was stained by HE staining. Myeloid tissue disorganization was obviouse, and the number of karyocyte was decreased. General condition was bad in mice. All mice were death in a week in model and calcium sensing receptor blocking CdCl2groups. After hUCB-MNCs transplantation, PB hemogram results show that the number of platelet was down to lowest at radiation damage14d, then began to slowly recover at21d in PB of hUCB-MNCs group mice. The number of neutrophil was down to lowest at radiation damage7d, then keeps to14d and began to slowly recover at21d. The number of white blood cells, lymphocytes and monocytes all were down to lowest at radiation damage7d, and began to recover at14d. But neutrophils, white blood cells and lymphocyte transiently recovered to pre-irradiation level, then again decreased. After the number of red blood cell transiently descend, and again recovery to normal level. But the number of neutrophils, platelets, white blood cells, lymphocytes and monocytes did not uncomplete recovered to pre-irradiation damage level. Compared with model group, the pemillage of Lin-Sca-1+c-kit+and CD133+flk-1+cells were significantly increased in PB and bone marrow (BM). The percentage of CDllb+, Gr-1+cells also was increased. On the base of hUCB-MNCs transplantation, Rgl low and high dose was administrated. Specaily in Rgl high dose group, the number of neutrophil, platelet and white blood cells all recovered to nomal level at cells transplantation in radiation damage mice. Lymphocytes and white blood cells began to recover at14d and21d, respectively. Compared with MNCs group, above indexes less descended, especially in Rgl high dose, the recovery of neutrophil and platelet were better, and duration of recovery was obviously in advance. Bone marrow karyocytes were distinctly increased. All mice were well survival. The pemillage of LSK in BM and CD133+flk-1+cells were higher than MNCs and control groups. The percentage of CD11b+cells was higher than model group. The percentage of Gr-1+cells was higher than model and MNCs groups.The percentage of Ter-119+cells was highest in model mice, and Rgl groups were higher than MNCs group. Bone marrow karyocytes were higher in Rgl high dose than in MNCs group. Effects of CaSR agonist GdCl3on PB hemogram, the pemillage of LSK and CD133+flk-1+cells were similar to Rgl high dose group in BM and PB. The levels of [Ca2+]i were increased by concentration dependent method with Rgl0.1-3μmol/L in bone marrow karyocytes from irradiation damage mice with hAMSCs transplantation.At hAMSCs transplantation4w, anti-human cell nucleus antibody-FITC labeled cells were observed that diffused distribution in myeloid tissue under fluorescence microscope. Except for relatively less MAB1281-FITC positive cells in Rgl low dose, there are not obviouse difference among MNCs, Rgl high dose and GdCl3group.Conclusion Rgl obviously improved recovery of hematopoiesis function in irradiation damage SCID mice with hUCB-MNCs transplantation. Rgl significantly decurtated recovery duration of neutrophil and platelet reaching normal level, and so blood cells. Effect of Rgl high dose were similar to CaSR agonist GdCl3on the recovery of hematopoiesis function in irradiation damage SCID mice. This effect is related to increasing [Ca2+]i level in bone marrow karyocyte, and to heighten proportion of HSCs and EPCs.