Stimulating effect of catechin, an active component of Spatholobus suberectus Dunn, on bioactivity of hematopoietic growth factor

Background Hematopoietic growth factor （HGF） is indispensable to hematopoiesis in the body. The proliferation and differentiation of hematopoietic cells must rely on the existence and stimulation of HGF. This study investigated the effect of catechin, an active component extracted from Spatholobus suberectus Dunn （SSD）, on bioactivity of granulocyte-macrophage colony-stimulating activity （GM-CSA）, burst-promoting activity （BPA） and megakaryocyte colony-stimulating activity （MK-CSA） in spleen condition medium （SPCM） of mice to clarify the hematopoietic mechanism of catechin and SSD. Methods Spleen cells of mice were separated and spleen condition medium （SPCM） was prepared from spleen cell culture. Bone marrow cells of mice were separated and cultured in a culture system including 10% （v/v） SPCM （induced by catechin in vivo or ex vivo） for 6 days. Granulocyte-macrophage colony forming units （CFU-GM）, erythrocyte burst-colony-forming units （BFU-E） and megakaryocyte colony-forming units （CFU-Meg） formation were employed to assay the effects of different treatment on the bioactivity of GM-CSA, BPA and MK-CSA in SPCM. Results SPCM induced by 100 mg/L catechin ex vivo could promote the growth of CFU-GM, BFU-E and CFU-Meg, which indicated that catechin could stimulate the production of GM-CSA, BPA and MK-CSA in SPCM. SPCM prepared at the fourth day of spleen cell culture showed the best stimulating activity. The bioactivity of GM-CSA, BPA and MK-CSA in the SPCM prepared after intraperitoneally injecting catechin into mice was also increased. The number of CFU-GM, BFU-E and CFU-Meg gradually increased as the dose of catechin increased and the time of administration prolonged. CFU-GM, BFU-E and CFU-Meg of the high-dose catechin group were significantly higher than those of the control group （P〈0.01） and reached the maximum at the seventh day after administration. Conclusions This study suggests that catechin extracted from the active acetic ether part of Spatholobus suberectus Dunn can regulate hematopoiesis by inducing bioactivity of GM-CSA, BPA and MK-CSA in SPCM of mice. This may be one of the mechanisms for the hematopoietic-supportive effect of catechin and Spatholobus suberectus Dunn.

Effects of Chinese Herbal Compound on the Blood Physiological Indices and Cytokines of Myelosuppressive Mice

[ Objective ] The research aimed to study the effects of Chinese herbal compound on the blcod physiological indices and cytokines of myelosuppressive mice. [ Metho] Myelasuppressive mice model was established by intraperitoneal injection of 80 mg/kg cyclophosphamide. Chinese herbal compound was composed of Houttuynia cordata, Taraxacum mongolicum, Citrus reticulata peel, Atractylodes chinensis, Paeonia sterniana, Atractylodes macrocephala and Angelica sinensis. The effects of Chinese herbal compound and Astragalus polysaccharide at different doses on the blood physiological indices and hematopoietic growth factors of myelosuppressive mice were discussed. [ Result] Myelosuppressive mice medel was successfully established. The total white blood cell count, total nentrephile granulocyte count, total lymphocyte count, total platelet count and the contents of serum interleukin-6 and erythropoietin in myelosuppressive mice were significantly decreased. The total erythrocyte count, the contents of hemoglobin and imerleukin-3 were decreased, without significant difference. The blood physiological indices and the contents of interleukin-3 and erythropoietin in myelosuppressive mice could be improved by intragastric administration of Astragalus polysaccharide and Chinese herbal compound at different doses for 3 days or 7 days. The effect of 20 g/kg Chinese herbal compound was the best after administration for 3 days, and the effect of 10 g/kg Chinese herbal compound was the best after administration for 7 days. The total white blcod cell count, total neutrophile granulocyte count, percentage of neutrephile granulocyte, total lymphocyte count, total count of middle cells, percentage of middle cells, total platelet count, contents of interleukin-3 and erythropoietin in myelosuppressive mice could be extremely significantly improved by intragastric administration of 10 g/kg Chinese herbal compound for 7 days（ P 〈 0. 01 ） , and the reduction of red blood cell count, hematecfit, the contents of hemoglobin and interleukin-6 induced by cyclophesphamide could be inhibited （P 〈 0.01 ）. [ Conclusion ] Chinese herbal compound could improve the hematopoietic function of myclosuppressivc mice induced by cyclophosphamide and its effect was better than Astragalus polysaccharide. 20 g/kg Chinese herbal compound reacted fast and the reaction of 10 g/kg Chinese herbal compound was slow ,but its efficacy was lasting.

Hematopoietic-supportive effect of (2S, 3R)-ent-catechin on marrow-depressed mice

Hematopoiesis is an active process of cell proliferation, differentiation andrelease. It is the process during which hematopoietic stem cells (HSCs) proliferate anddifferentiate to mature blood cells under the effect of hemetopoietic growth factors (HGFs) incertain hematopoietic microenvironment. HSCs are sources of hematopoiesis of a body that canself-renew, differentiate to blood cells of every lineage and maintain the constancy of them. As themajor tissue of hematopoiesis bone marrow is filled with all kinds of blood cells in variousdevelopmental stages. Under the normal conditions, the ordered proliferation and differentiation ofhematopoietic stem cells/hematopoietic progenitor cells ( HSC/HPC) depend on the regulation ofcytokine network. In present, Spatholobus suberectus Dunn (SSD), a traditional Chinese herb medicinethat has been used to invigorate the blood circulation for thousands years, is widely used torebuild blood after chemotherapy and radiotherapy and to treat hematopoietic diseases. Previousreports from our laboratory indicated that (2S,3R)-ent-catechin, epi (2S, 3R )-ent-catechin, gallic( 2S, 3R )-ent-catechin, et al extracted from SSD all could stimulate the proliferation of HPC inmarrow depressed mice, among which (2S, 3R)-ent-catechin obtained from spatholobus genus for thefirst time and whose content was the maxim showed the best effect. In this study, firstly, theeffect of (2S,3R)-ent-catechin on the quality and quantity of HSCs were assayed by detectingCD_(34)^+ expression. Secondly, we investigated the variation of cytokine (GM-CSF and IL-6)sero-level in mice treated with (2S,3R)- ent-catechin by ELISA. And IL-6 mRNA and GM-CSF mRNAexpressions induced by (2S,3R)-ent-catechin were detected simultaneously by RT-PCR technique inorder to clarify its mechanism.