The Mechanism Of Saponin Of Pearsonothuria Graeffei On Hematopoiesis In Myelosuppression Mice

The saponins are the major secondary metabolites, a kind of triterpenoids in the body wall of sea cucumber. In this study, the saponin of Pearsonothuria graeffei was first used to investigate acute toxicity in mice, 30 days oral toxication and deleterious effect on the growing development in rats, for the aim of finding its characteristic, target organ, and harm caused by long-time intake.According to the function characteristics and median lethal dose(LD50), the saponin of P. graeffei dissolved by 0.5% carboxymethylcellulose with distilled water was used to determine the hematopoiesis and mechanisms in bone marrow of myelosuppression mice which were induced by triple intraperitoneal injection of cyclophosphamide(Cy). Results are shown as follows:The LD50 of saponin of P. graeffei is 1.08 g/kg in male mice and 1.1g/kg in female mice. The saponin of P. graeffei has no remarkable effect on the contents of blood urea nitrogen(BUN), creatinine(Cr), aspartate aminotransferase(AST) and alanine transaminase(ALT) of serum in mice which were given a single saponins of P. graeffei, which indicated that there was no obvious damage to kidney and liver in mice. The rats grew normally and had shiny pelage during the 30 days oral toxication experiment. Also, the saponin of P. graeffei increased the number of white blood cell(WBC) and platelet(PLT) in peripheral blood, which indicated the saponin of P. graeffei had the hematopoiesis function, and the saponin of P. graeffei had no obvious damage to the liver, kidney and cardivascular and so on according to investigating the biochemical index of blood and urine in rats.The hematopoiesis function was studied by cell and tissue culture. It was proved that the saponin of P. graeffei could enhance the number of peripheral blood WBC, red blood cell(RBC), PLT, hemoglobin(Hb), reticulocyte(RET) and bone marrow nucleated cell(BMNC), promote the proliferation and differentiation of hemopoietic stem cells (HSC), increase the number of CFU-GM(P<0.01), CFU-E(P<0.05), and CFU-Meg(P<0.01) and stimulate secretion of sorts of cellular factors to promoting hematopoiesis function of bone marrow in mice.The effects of saponin of P. graeffei on cell cycle and apoptosis in mice.were investigated using flow cytomety and RT-PCR. The results showed that the saponin of P. graeffei promoted hematopoiesis by regulating cell passing S phase to G2/M phase, increasing the mRNA expression of cyclinB1, bcl-2(P<0.01) and bcl-xl(P<0.01), enhancing cell mitosis, supressing cell apoptosis。The mRNA expressions of GM-CSF, EPOR and TPO were investigated by RT-PCR. The results showed that the saponin of P. graeffei promoted hematopoiesis function according to evidently increasing mRNA expression of GM-CSF(P<0.01), EPOR (P<0.05), TPO(P<0.01) and the synthetize and secretion of EPO, GM-CSF and TPO, promoting EPO, GM-CSF or TPO binding to its receptor effectively.The safety evaluation of saponin of P. graeffei was first investigate, which provides certain scientific proof while applying to clinical tests, nourishment and health care products in the future. The promoting hematopoiesis and mechuanism of saponin of P. graeffei was studied on the basis of security application, which provided theoretical basis on the development and high-value use of saponin of P. graeffei.