| Objective:To establish a mouse aplastic anemia model by hydracetin, X-ray and cyclophosphamide combination. To evaluate the function of the nutrients mixture to promote hematopoietic stem cell proliferation and hemoglobin synthesis in aplastic anemia (AA) mouse.Method:100 BALB/c type mice were fed and divided randomly into three groups: normal control group, AA model group and the different dose of the nutrients mixture groups. The mouse AA model was established by hydracetin, X-ray and cyclophosphamide combination. The normal control group was established by lead shielding irradiation alone and the saline injection. After the seventh day, mice were fed by nutrients mixture in a dose-dependent manner (1445.55, 963.7, 674.59 mg / kg ? d). The mice were killed by cervical dislocation and then were determined by blood, bone marrow, mitochondria of hematopoietic cells, hematopoietic stem cell colony formation and liver, spleen pathological changes after 45 days. Results:1 On the 7th day, mice fell into a decline including gloomy tousled fur, slow acting, less eating and blunt reaction compared to the control group. Furthermore, the status of mice was improved when the nutritious composition was increased in each group. But the mice were deteriorated in the aplastic anemia group. Meanwhile, the weight of mice was decreased in the aplastic anemia group and increased in the different nutritious groups(*p<0.05). 2 Compared to the control group, the peripheral hemogram of mice in the model group showed a remarkable decrease (*p<0.01). The peripheral blood cells were increased significantly in each of the high, middle, and low nutritious composition group compared to the aplastic anemia group (*p<0.01). Peripheral blood smear indicated that neutrophil was decreased more obviously than that of the whole whit cell population, but the ratio of the lymphocytes was increased relatively. However, there was no significant difference between the each nutritious supplying group and the control group (*p<0.05). The erythropoietin (EPO) of blood in AA was higher than the control group, and was negatively correlated with RBC and Hb. The EPO was a significant reduction by compensatory role (*p<0.01), with a dose dependent.3 The number of nucleated cells in bone marrow exhibited decreased significantly compared with controls (*p<0.01). The nucleated cells of bone marrow in nutrients mixture group were significantly higher than that of AA model (*p<0.05). Transmission electron microscope showed that the number of mitochondria in similar cells was markedly increased in nutrients mixture groups, comparing with AA group, (* p <0.01).4 Colony formation assay showed that the colony formation exhibited decreased significantly compared with controls. The colony formation rates of nutrients mixture groups were increased markedly, comparing with AA group (* p <0.01), with a dose dependent.5 Liver, spleen biopsy showed that the liver tissue hydroplasia obviously and the lobular structure was less clear, the liver sinusoids and central venous were expanded and congestive; the spleen tissue was loose and the number of spleen body decreased. The liver tissue edema was reduced and lobular structure was clear in nutrients mixture groups, comparing with AA group. Furthermore, the number of splenic body was also increased in nutrients mixture groups.Conclusion:1 The method to establish aplastic anemia model suggested that the pathological changes of peripheral blood and bone marrow were persistent and stable, and consistent with clinical situation of aplastic anemia. This method was simple,and had a high achievement ratio. It could be used to screen relative drug.2 Nutrients mixture can play a crucial role. It improves the bone marrow damage of AA mice, promotes blood cell production and hematopoietic stem cells proliferation, as well as increases the number of mitochondrial. Moreover, the nutrients mixture recovers liver, spleen damage of AA mice.
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