| Background and Objective:Aplastic anemia, an unusual hematologic disease, is the paradigm of the human bone marrow failure syndromes. It can be attributed to "consumptive disease" or "emia" according clinical syndrome, although there is no name of "aplastic anemia" during traditonal Chinese medicine (TCM). Now more and more researchers pay attention to its pathogenesis of "Kidney deficiency" and "haemostasis". For many years, Yi Shen Hua Yu decoction has been used for the promotion of "blood production" and "tonifying kidney" and has shown favorable effect on hematopoiesis, which pharmacological mechanism may be considered through improving hemopoietic microenvironment, modulating the positive hematopoietic factors and suppressing the apoptosis of hematopoietic stem cell. However the existing research does not explain all of the mechanism on hematopoiesis.It is close of relationship between entrails in basic theories of Traditonal Chinese Medicine such as the kidney stores the essence, essence birth marrow, marrow birth blood and marrow filling brain. It is close relationship during "emia", "nervous system disease" and kindey deficiency. So hematopoietic system disease may share the same pathogeny of kidney deficiency with nervous system disease.5-Hydroxytryptamine (5-HT, Serotonin) which is an indolamine compounds, plays an important role between central and peripheral organs. Since Nemecek et al.firstly reported that 5-HT has positive effect on the growth of vascular smooth muscel cells, then many researchers confirmed its growth promoting effects on numerous types of cell including fibroblasts, nerve cells, neuroglial cells, osteoblasts, mesangial cells, epithelial cells and megakaryocyte. Other reserches indicated that central regulation of hemopoietic stem cell is mediated by different neurotransmitter mechanisms:erythropoiesis depends on the state of serotoninergic structures, while granulocytopoiesis is regulated both by the dopamine-and serotoninergic systems. In 1996, Yang et al reported that serotonin exerted mitogenic stimulation on megakaryocytepoiesis mediated by 5-HT2R, recently they demonstrated that serotonin could significantly expanse of human cord blood CD34+cells to early stem/progenitors and multilineage committed progenitors (BFU/CFU-E,CFU-GM,Mk-CFU), enhance the colony-forming unit-fibroblast (CFU-F) formation, and antiapoptosis of megakaryoblastic M-07e cells in vitro.On the other hand, it has been found that herbal of activating blood and reinforcing kidney medicine can modulate the 5-HT secretion of central nervous system and improve the learning and memory function of vascular dementia rat model, as well as its favorable effects on hematopoiesis of anemia patients, so we infer that one mechenism of Yi Shen Hua Yu decoction (YSHY) on hematopoiesis might through regulate the 5-HT secretion of periphery blood and modulating the expression of 5-HT receptors on bone marrow mononuclear cells. We have successfully make the acute myelosuppression rat modle and have proved that YSHY decoction could improve hematopoiesis, relieve bone marrow stromal edema and elevate the concentration of plasma 5-HT in myelosuppression rat modle. Now we further investigate the role of YSHY decoction on megakaryocytopoiesis through 5-HT2AR pathway, in order to nourish the mechanism of our traditional Chinese medicin on megakaryocytopoiesis.Methods:32 SD rats were randomly divided into normal control group, model control group, Yi Shen Hua Yu decoction (YSHY) group and YSHY decocotion+ketanserin (ketanserin 2mg/kg.d) group. There were 8 rats in each group.5-Fluorouracil and Busulfan were used to make the acute myelosuppression rat modle according reference. Intraperitoneal injection at the dosage of 150mg/kg were given to the rats at days 0, busulfan was administrated by gastric perfusion at days 5, days 12, and days 19, then Yi Shen Hua YU decoction was adminestrated by gastric perfusion daily for 16 days at the dose of 0.2ml/10g(2g/ml), with or without ketanserin(2mg/kg), while the model and normal control group were treated with drenched sodium chloride at the same dose and volume, then bone marrow mononuclear cells (BMMC) were counted and CFU assays were used to determine the ettects of Yi Shen Hua YU decoction on the megakaryocyteic progenitor cells and other lineages. Last analyses the expression of CD41 and 5-HT2AR were conducted in bone marrow monocyte by flow cytometry.Result:(1) The number of BMMC count:YSHY decocotion could significantly elevate the count of BMMC in model rats, and this effect could be suppressed by ketanserin in vivo.(2)Mk-CFU and mix-Mk-CFU assays:YSHY decoction could elevate the count of megakaryocyte colony forming unit (MK-CFU) and mix-megakaryocyte colony forming unit (mix-MK-CFU) in model rats, and this effect could be suppressed by ketanserin in vivo.(3)GM-CFU assays:YSHY decoction could elevate the count of Granulocyte monocyte colony forming unit (GM-CFU), while this effect could not be suppressed by ketanserin in vivo.(4) Flow cytometry analysis:YSHY decoction could upregulate the CD41,5-HT2AR expression on the BMMC of model rats, while ketanserin only suppressed CD41 expression in vivo.Conclusion:(1) YSHY decoction might promote megakaryocytopoiesis on myelosuppression rat model through enhancing 5-HT2AR expression of BMMC. (2) ketanserin might selectivly suppress the effect of YSHY decoction on megakaryocytopoiesis. |
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