| Objects:To observe the influence of Buxue Yimu Pill on hematopoiesis and immune system in mice with qi and blood deficiency,and to explore the mechanism of qi and blood double supplement,which is of great significance for its clinical application,research and development.Methods:Four weeks healthy 60 male kunming mice were randomly divided into 6groups according to the random number table: normal control group,model control group,positive control group of traditional Chinese medicine,positive control group of western medicine,and low and high dose group of nourishing blood and benefiting mother pill(10 mice in each group).Except the normal control group,all the other groups adopted "orbital blood collection and hemorrhea,and intraperitoneal injection of cyclophosphamide every other day" to replicate the double-deficiency model of qi and blood for 9d.After successful modeling,the blank control group and the model control group were given normal saline.The positive control group was given siwu decoction(0.24g/ person),the positive western medicine group was given ferrous lactate oral liquid(0.2m L/person),and the blood and yimu pill group was given low and high doses of blood and yimu pill liquid.Low-dose gavage was 1.82g/kg·-1 while high-dose gavage was 7.28g/kg·-1.Continuous gavage for 10 days.After the last administration,the morphological signs of mice were observed,and the indexes of peripheral blood,related viscera index and the number of nucleated cells in bone marrow were measured.Hematoxylin-eosin staining(HE staining)was used to detect spleen damage.Whole blood was collected by 1.5m L EP tube.The serum was centrifuged after standing at room temperature for 3h.-80℃ refrigerator storage.One serum was used for the determination of metabolomics,while the other serum was used for the determination of erythropoietin(EPO),granulocyte-macrophage colony stimulating factor,tumor necrosis factor-alpha(tnf-alpha),interleukin-2(il-2)by ELISA.Results:1.Morphological observation of each group of mice before the last dose.Compared with the normal group,most mice in the model group were curled up and inactive.Its fur was withered and dull and yellow.The physiological state of the mice treated with buxue yimu pill was not significantly different from that of normal mice.2.The weight-bearing swimming test of mice showed that the weight-bearing swimming time of the mice after drug administration was significantly different from that of the model group(p < 0.05),and there was no significant difference from that of the normal group(p > 0.05).3.Peripheral blood imaging results showed that the WBC,RBC,HGB and PLT Numbers of mice in the model group were significantly different from those in the normal group(p < 0.05).However,there was a significant difference between the peripheral blood of mice treated with buxue yimu pill and the model group(p < 0.05).4.Compared with model group,give the female pill in mice blood interest the blood lactic acid,blood lactic acid,BLA)content and blood urea nitrogen(blood urea nitrogen,BUN)content exist significant difference(p < 0.05).5.The number of nucleated cells in the model group was significantly different from that in the normal group(p < 0.01).However,there was no significant difference in the number of nucleated cells between the mice treated with buxue yimu pill and the mice in the normal group,and there was a significant difference with the mice in the normal group(p < 0.05).6.The results of HE staining of the spleen of mice showed that the spleen of the normal group had clear medulla,thicker cortex and higher lymphocyte density.In the model group,the cortical medulla was not clear and the cells were sparse.The red and white boundaries of spleen and medulla were clear and the cell density was higher in mice with low and high doses of buxue yimu pill.7.Metabonomics,according to the results of blood benefit female pill dosage group and positive medicine group were between model group and normal group,which shows that after treatment with drugs,three groups of mice serum metabolite levels tend to be normal development,also found that compared with positive group,blood benefit female bolus dose group on the metabolites level recovery ability strong positive contrast group and positive medicine group and western medicine positive group of western medicine has good effect to restore to the normal groups.8.There was no significant difference in serum gm-csf and il-2 levels in mice of blood-enriching and mother-nourishing pills compared with the normal group(p >0.05),but significantly increased compared with the model group(p < 0.05).Serum levels of EPO and TNF-alpha in mice of buxue yimu pill were significantly lower than those in the model group(p < 0.05),and there was no significant difference compared with those in the normal group(p > 0.05).Conclusion:1.Buxue yimu pill can improve the peripheral blood image of mice with dual deficiency of qi and blood,improve the signs of dual deficiency of qi and blood,repair the spleen tissue of mice,and play its role in the treatment of dual deficiency of qi and blood.2.GC-MS metabolomics technology combined with metabolic pathway regulation was used to analyze the potential biomarkers in the serum of mice,and the changes of these metabolites were involved in the body’s fatty acid metabolism,amino acid metabolism,energy metabolism,etc.After the treatment of nourishing blood and nourishing mother pill,the level of some metabolites was closer to the normal group.3.The mechanism of the effect of nourishing blood and yimu pill on treating qi and blood dual deficiency syndrome may be related to the stimulation of hematopoietic function of bone marrow and the regulation of immunity.By improving the content of gm-csf and EPO in serum,bone marrow hematopoietic can be stimulated.The therapeutic effects of TNF-a and IL-2 in serum were studied. |
