| Blood deficiency (BD) is a common TCM syndrome in clinic, which is often caused by insufficient origin of blood enriching, excessive blood losing, kidney-essence deficiency, etc. Si-Wu-Tang (SWT) that consists of Rehmanniae Radix, Angelica Radix, Paeoniae Radix, and Chuanxiong Rhizoma is a well-known Chinese formula widely used in clinic for enriching blood, activating blood, regulating menstruation, etc. But up to now, the functional mechanism of SWT has not been totally understood. This doctoral thesis focuses on the experiments in vivo and in vitro to study the radiation effect on protein expressions of mouse bone marrow cell, the blood enriching function of Si-Wu-Tang and its active monomers on blood deficiency mice induced by irradiation, and the effect of the medicines on the protein expression of bone marrow cell and mRNA expression of hematopoiesis related cytokines of the mice. The experiment also researches the effects and mechanisms of Si-Wu-Tang, paeoniflorin and ligustrazine on proliferation and protein expression of human bone marrow stromal cell respectively. The observations could provide a view in better understanding the hematopoietic molecular mechanisms of irradiation injured bone marrow and that of blood enriching function of Si-Wu-Tang and its active monomers. The experiments can be summarized as the following. 1. The study of blood enriching function of Si-Wu-Tang and its active monomers on blood deficiency mice induced by radiation. With the methods of peripheral hemogram measurement and myeloid colony cell culture, the blood enriching functions of Si-Wu-Tang, paeoniflorin, ligustrazine, ferulic acid and fructose on blood deficiency mice induced by 3.5Gy 60Co irradiation were studied. The results demonstrated: (1) After 3.5Gy 60Co γ-ray irradiation, peripheral leucocyte count of C57BL/6J mice declined significantly, the colony number of CFU-GM, CFU-E, BFU-E and CFU-mix cultured in vitro decreased significantly, and the thymus and spleen index declined significantly. It is postulated that after irradiation, the hematopoietic function of mouse bone marrow was inhibited and the immune organs and immune function of the mice were injured; (2) Si-Wu-Tang and its active monomers, paeoniflorin, ligustrazine, ferulic acid and fructose, could all promote the increase of peripheral leucocyte numbers and hematopoietic progenitor cell colony numbers of bone marrow of blood deficiency mice. Here, SWT has most continual role. The formula and fructose have most significant effect, and paeoniflorin takes second place; (3) Fructose and ligustrazine could respectively promote recovery of atrophic thymus and spleen induced by irradiation of mice and SWT had same effect for recovery of both organs. It can be concluded that SWT and its active monomers have the function of enriching blood and promoting immune organ weight recovery to blood deficiency mice induced by irradiation. Fructose and paeoniflorin may be the main active compositions of SWT that exert blood enriching function and fructose and ligustrazine may be the main active compositions of SWT that improve immunological function. 2. The effect of radiation on protein expression of mouse bone marrow cell and the effects of SWT, paeoniflorin and ligustrazine on the expression change With the proteomic approaches, such as two-dimensional electrophoresis (2-DE), gel dyeing, image analysis, in-gel digestion, mass spectrometry, and database search, the research investigated the effect of radiation on protein expression of mouse bone marrow cell and the effect of SWT, paeoniflorin and ligustrazine on protein expression of blood deficiency mice induced by irradiation. The experiment results showed that (1) (2) Radiation could induce many proteins of mouse bone marrow cell to change. Including (i) radiation directly or indirectly acted on the signal molecules in signal pathway of hematopoietic growth factors. Through inhibiting protein and mRNA of ras, MAPKK mRNA and LSP1 expressions and increasing Grb14 and HCP expression of mouse bone marrow cell, the ra...
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