Research On Immunological Activity Of Traditional Chinese Medicine Venenum Bufonis In Vitro

Nowadays,studies have confirmed that activity of traditional Chinese medicine Venenum Bufonis has immunity regulated function, anti-virus, anti-tumor, local anaesthesia, boost pressure, et al. Venenum Bufonis is becoming more and more important in clinical application because of its diphasic actions of anti-tumor and immunity regulated function and other actions. But the chemical components of Venenum Bufonis are complicated, immunological active sites and chemcial constitution of active components are not obvious. These cause many difficulties to research the immunological activity of Venenum Bufonis and are especially not beneficial to study the pharmacological mechanism in molecular level, it is difficult to control administration in clinic and pharmacodynamic reproducibility is poor. Recently, the studies of immunological activity of Venenum Bufonis preparations and crude extracts only stayed at the level of detecting the function index of immunologic system, and did not expound the mechanism of immunologic enhancement in molecular level.Thus, in the following experiments, we extracted different solvent component of Venenμm Bufonis, then screened the extracts with immunological activity detection and studied the immunoregulatory effects of Cinobufagin which come from the chloroform solvent component of Venenum Bufonis in vitro.MTT assay was used to detect the proliferation and conversion of the splenic lymphocytes of mice. Metaboilic level and phagotrophic activity of PMΦwere also measured. Results showed that the extracts of Venenum Bufonis VB2,VB4,VB6 combined with Con A could promote the proliferation and conversion of the splenic lymphocytes of mice, and also could enhance the metaboilic level and phagocytosis function of PMΦ; We first confirmed that Cinobufagin has the effect of immunological enhancement. Cinobufagin administered only and combined with Con A or LPS could promote the proliferation and conversion of the splenic lymphocytes of mice, and also could enhance the metabolic capability of MTT and the phagocytosis function of peritoneal macrophage. The anti-tumor activity of Cinobufagin has been reported at home and abroad, so it implied that Cinobufagin may have diphasic actions of anti-tumor and immunity regulated function. The percentage of every CD4,CD8 subgroup of T lymphocytes and S phase cells of the splenic lymphocytes of mice were detected by Flow cytometry. According to the result, we first expounded the mechanism of immunologic enhancement of Venenμm Bufonis in molecular level. It also showed that VB2, VB4, VB6 and CBG had immunoenhancement effects on the immunologic function of mice in vitro which was related to increase the percentage of DP subgroup of T lymphocytes and S phase cells.According to the experiments, the researsh explored the influence of Venenum Bufonis on immunological function in vitro. The results of this study will benefit to the further research on the immunoenhancer of Venenμm Bufonis.