| Objective: Dahuangzhechongwan(DHZCW) has effects of promoting blood circulation to remove blood stasis, invigorating deficiency gradually, treating the deficiency of emaciated abdominal stuffiness, skin dry and no nourishment, darkish for eyes, and drying blood in body. In clinic, DHZCW were widely used to treat diseases correlation with immunization disorder, such as that of the liver, malignancy, chronic malignancy, dryness syndrome, arthropathy, skin disease, hyperplasia of mammary glands, endometriosis et al. The pharmacology of this prescription have been widely researched, such as resisting clotting, haemalysis, thrombolytic, improving hemorheology, protecting liver, resisting liver from fibrosis, bating nephrosis and inhibiting the fiber proliferation of kidney. So the purpose of this research is to study the immune regulation mechanism of DHZCW with modern immune technology and molecular biology knowledge. This research can reveal the pharmacology of DHZCW and provide theoretical base for reasonable medication in clinical. By the study, new thoughts and methods for the research on the essential of blood-activating and stasis-γemoving prescriptions can be provided.Methods: In order to reveal the immunological mechanism of DHZCW, The effect of DHZCW on the immune organ and molecule of normal mice and immune dysfunctional mice were studied. Just as following : (1)To study the effect of DHZCW on ponderance index and structure of thymus and spleen of normal and immune dysfunctional mice by detecting the structure of immune organ by optical microscope and electronical microscope, (2)the effect of DHZCW on double-positive CD4+CD25+T lymphocyte subpopulation in the peripheral blood and the cell suspension by flow cytometry. (3)To study the effect of DHZCW on the concentration of IL-4, IFN-γ in peripheral blood and spleen cell culture supernant in each group mice by ELISA.(4)To study the expression of IL-4mRNA,IFN-γ mRNA in thymus and spleen tissue and spleen cells of each group mice were detected by the hybrindization in situ respectively.Results: (1)The effect of DHZCW on the ponderance index and the structure of thymus and spleen of normal mice and immune dysfunctional mice: Obviously shrink and regression in the thymus and the spleen appear in the immunospression mice groups. The structure were blur and the cells in immune organ decreased and diffluenced, chromat becomed fade. Given different dosages DHZCW differently lessen the variety of these mice. The indicate that DHZCW affected the immune organs. And the thymus and the spleen of ponderance index showed no effection in each group.(2)the effect of DHZCW on the subtype of T lymphocyte cell in the peripheral blood and the cell suspension: the rate of CD4+CD25+T cell in the peripheral blood and the cell suspension of the immunsuppression group was higher than the normal group, DHZCW had no effect on the mice in the normal group, but had reduced on the mice in theimmunosuppression group.(3) The concentration of IL-4,IFN-γ in peripheral blood and spleen cell culture supernant by ELISA appeared: no difference in normal groups. IFN-γ apparently decreased, while IL-4 apparently increased in hyperimmunity model groups. After DHZCW, the effects of IFN- Y apparently increased markedly, while IL-4 decreased. Both IFN- Y and IL-4 apparently decreased compare to the normal mice in immunsuppression groups, there was obvious difference reduced in IFN- Y contents and no difference in IL-4 after treatments.(4)The results of in situ hybrindization showed: no difference in normal group, and increased expression of IFN- Y mRNA no difference in IL-4mRNA of immune organs in immunsuppression mice. For the hyperimmunity, DHZCW can enhance the expression of IFN-γ mRNA and reduce IL-4mRNA.Conclusion: The study did experiment research on the immunomodulation of DHZCW from the aspect of formal structure, immuological cell, and protein. DHZCW can improve the stucture of immune organ, it has the immune function of adjustion . The regulation was carri... |
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