Effect Of Total Saponinis Of Panax Ginseng On The Expression Of Erythropoietin Receptor In Hematopoietic Cells And K562 Cell

Total saponins of panax ginseng (TSPG) is one of the main effective ingredient extracted from Panax Ginseng which plays a vital role in"invigorating qi and promoting haematopoietic".Our lab previous study showed that: TSPG can promote erythro-progenitor cells (BFU-E, CFU-E) differentiation and proliferation, and promoting K562 leukemia cell apoptosis, inhibite their proliferation, but its molecular mechanism remains unclear. erythropoietin receptor plays an important role in the haematopoietic cell proliferation and differentiation,and K562 cells anti-apoptosis and differentiation into erythroid cells, receptor bind with Eerythropoietin product signal transduction ( STAT5), finally launch related gene transcription. In order to discover the molecular biology mechanism that which promote erythroid hemocytopoiesis and inhibit the K562 cell proliferation . this study use umbilical cord hematopoietic cells and K562 as experiment object,add diversity concentration TSPG in culture system to induce it for 24h, investigate TSPG effecton EpoR expression of both. Now two part of the brief as follows:1.TSPG effect on the EpoR expression of umbilical cord hematopoietic cells: Separate umbilical cord mononuclear cells , subgroup and induced with different concentrations TSPG (0,25,50,75,100 mg / l) for 24 h, use flow cytometry,Cell ELISAdetect the quantity of EpoR on cell surface; cytoplasm ELISA detect the cytoplasm quantity of EpoR use Immunocytochemical stain to observe the positive rate of cell that express EpoR in smear that pre-induced by TSPG and post- induced . The results showed that: 50 mg/l group of compare with the 0 mg / l control group, the EpoR on cell mambrane surface increased significantly, the quantity of EpoR in cytoplasm has not changed, the positive rate of cell that express EpoR induced by TSPG in smear fall down than before induction. Conclusion: TSPG can induce the quantity of EpoR on umbilical cord hematopoietic cell mambrane decrease, the quantity of EpoR in cytoplasm has not increase,this maybe the one of mechanism that ginseng promote hematopoietic cell proliferation and induce it differentiate into erythroid cells.2. TSPG effect on the EpoR expression of K562 cells: culture K562 cells in vitro induced by different concentrations TSPG (0, 50, 100,200,300,500 mg/l) 24 h, use flow cytometry, cell ELISA detect the quantity of EpoR on cell surface; cytoplasm ELISA detect the quantity of EpoR in cytoplasm; laser confocal microscopy observe fluorescence image of K562 cell that induced by TSPG and not. The results showed that:up than 200 mg / l group than 0 mg / l group,the quantity of EpoR on the surface of the cell membrane significantly reduce, the quantity of EpoR in cytoplasm increase accompany with the TSPG concentration increasing, the fluorescence intensity on cell membrane of K562 cell that induced by TSPG than uninduced weaken. Conclusion: TSPG can induce the EpoR on the K562 cell membrane transferred into the cytoplasm, promote the quantity of EpoR on membrane to decrease.