A Comparative Study on the Effects of Different Processing Methods of Asini Corii Colla on Blood Cell Components and Related Cytokines in Mice with Blood Deficiency Syndrome
Objective To investigate the hematopoietic effects and mechanisms of Asini Corii Colla with different processing methods on blood-deficient mice.Specifically,we aim to evaluate the blood-nourishing properties of Asini Corii Colla extracted using high-pressure steam extraction compared to the traditional atmospheric pressure extraction method.Methods Seventy female C57BL/6 mice were randomly divided into the following groups:normal control group(Normal),model group(Model),Asini Corii Colla processed by atmospheric pressure method low dosage group(aL),Asini Corii Colla processed by atmospheric pressure method high dosage group(aH),Asini Corii Colla processed by high-pressure steam low dosage group(AL),Asini Corii Colla processed by high-pressure steam high dosage group(AH),and positive control group treated with vitamin B12(VB12).Except for the normal group,all other groups were induced to develop blood deficiency syndrome in mice by combining acetylphenylhydrazine(APH)with cyclophosphamide(CTX).After the treatment,whole blood,kidney,and femur tissue samples were collected from the mice.The peripheral blood parameters of the mice were determined using a hematology analyzer.The levels of hematopoietic cytokines in the mouse serum were measured by ELISA.The protein levels related to hematopoietic function in the mouse serum and kidney tissue were determined by Western blot.Hematoxylin and eosin(HE)staining was performed to observe the pathological changes in the bone marrow tissue.Immunohistochemistry was used to evaluate the protein expression levels of granulocyte colony-stimulating factor(G-CSF)and granulocyte-macrophage colony-stimulating factor(GM-CSF)in the bone marrow tissue.Results Asini Corii Colla processed by different techniques significantly increased the levels of peripheral blood leukocytes,erythrocytes,hemoglobin,and lymphocytes in mice with blood deficiency.It also decreased the mean corpuscular volume and mean corpuscular hemoglobin concentration in peripheral blood,effectively improving the symptoms of reduced hematopoietic cells and increased adipose tissue in mice with blood deficiency.It increased the number of nucleated cells in the bone marrow,enhanced the protein levels of G-CSF and GM-CSF in the bone marrow,and increased the levels of erythropoietin(EPO),interleukin-3(IL-3),and GM-CSF in the serum.It also increased the expression of EPO in the kidneys and decreased the levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the serum.Compared with Asini Corii Colla processed by traditional atmospheric pressure extraction,the Asini Corii Colla produced by high-pressure steam extraction exhibited more significant effects in increasing blood cell counts in peripheral blood,increasing the number of nucleated cells in the bone marrow,enhancing the protein levels of G-CSF and GM-CSF in the bone marrow,increasing the levels of erythropoietin(EPO)and G-CSF in the serum,and decreasing IL-6 levels.Conclusion Asini Corii Colla processed by different techniques demonstrated significant blood-supplementing effects in mice with blood deficiency,with the high-pressure steam extraction technique showing stronger effects than the traditional atmospheric pressure extraction technique.The mechanism involved the upregulation of EPO,IL-3,GM-CSF,and G-CSF as positive regulators of hematopoiesis,increased expression of EPO in the kidneys,enhanced protein expression of GM-CSF and G-CSF in the bone marrow,as well as the downregulation of TNF-α and IL-6 as negative regulators of hematopoiesis.
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