Study On The Thrombopoietic Effects And Mechanism Of Total Saponins From Sanguisorba Officinalis L

Objective:To investigate the thrombopoietic effects of total saponins from sanguisorba officinalis and the effects on three important targets:TPO/TPOR-. IL-3/IL-3R、SCF/c-kit, which clears the mechanism of total saponins from sanguisorba officinalis’ thrombopoietic effects.Methods:Studied the thrombopoietic effects and mechanism of total saponins from sanguisorba officinalis L(TSSO) by both in vitro and in vivo experiments. In vivo tests included:Mouse were injected to cyclophosphamide to establish the animal model of thrombocytopenia. The TSSO(0.5、0.1mg/kg) was administrated. then the peripheral blood and the coagulation time was measure on the forth day, seventh day and ninth day. In vitro tests included:(1)The proliferation capacity of Baf3/Mpl cells were determinated by the methods of MTT.(2) The proliferation capacity of Baf3cells were determinated by the methods of CCK8.(3)The proliferation capacity of32D cells were determinated by the methods of CCK8.(4) The differentiation capacity of32D cells were determinated by the methods of Giemsa staining.(5)The effects of TSSO on the expression level of TPO receptor in Baf3/Mpl cells were determinated using RT-PCR.(6) The effects of TSSO on the expression level of c-kit in32D cells were determinated using RT-PCR.(7) The effects of TSSO on the expression level of c-kit in HEL cells were determinated using fluorescence probe method.Results:In vivo study showed that:TSSO can increase the number of the WBC, RBC and PLT. In vitro study showed that:(1)The proliferation of Baf3/Mpl cells were significantly suppressed in the absence of TPO. When cultured with TPO the Baf3/Mpl cells were significantly promoted. TSSO (10μg.ml-1) also increase the proliferation when cultured with TPO, and better than TPO alone. TSSO promotes alone proliferation of Baf3/Mpl cells.(2) The proliferation of Baf3cells were significantly suppressed in the absence of IL-3. When cultured with IL-3the Baf3cells were significantly promoted. TSSO (10μg.ml-1) also increase the proliferation when cultured with IL-3, and better than IL-3alone. TSSO promotes alone proliferation of Baf3cells, and presents the time dependent.(3)The proliferation of 32D cells were significantly suppressed in the absence of IL-3. When cultured with IL-3the32D cells were significantly promoted. TSSO (10μg.ml-1) also increase the proliferation when cultured without IL-3and presents the time dependent.(4)TSSO (40μg.ml-1) increase alone the number of the polyploidy megakaryocyte when exposed to32D cells for four days. And significantly increase alone the differentiation of32D cells and the number of the polyploidy megakaryocyte when exposed to32D cells for nine days.(5)TSSO(10μg.ml-1) increases alone the expression level of TPOR in Baf3/Mpl cells for48hours.(6) TSSO(10μg.ml-1) increases alone the expression level of c-kit in32D cells for48hours.(7) TSSO(20μg.ml-1) increases alone the expression level of c-kit in HEL cells for48hours.Conclusions:TSSO has its effect on promoting thrombopoiesis, and the mechanism is correlated to the up-regulation of TPO receptor and c-kit.