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《Pharmacology and Clinics of Chinese Materia Medica》 2016-06
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The research of Bufadienolides on immune function of mouse lymphocyte

Wei Xiaolu;Yang Jian;Wang Hongjie;He Jing;Bian Baolin;Institute of Chinese Materia Medica.China Academy of Chinese Medical Sciences;  
Objective: Observing the influence of bufadienolides effects on immune related cytokines in vitro and vivo,analyzing the immune system regulation,providing the pharmacological and pharmacodynamics evidence to clarify its immune mechanism. Methods: We aseptically isolated the mice spleen lymphocyte in vitro,prepared the lymphocyte suspension,and set different concentration groups. MTT method was used to detect mice spleen lymphocyte proliferation separately indured by concanavalin A( Con A) and lipopolysaccharide( LPS). Medicine was preincubated the lymphocytes for 6 h,then Con A or LPS separately was added to induce the lymphocyte activation,we used flow cytometry on cell supernatant to detect the cytokines of GM-CSF,TNF-α,IFN-γ,IL-1α,IL-2,IL-4,IL-5,IL-6 and IL-10,and evaluated its immunological effect. Set up animal model of As PC-1 pancreatic cancer in nude mice,after giving medicine for a certain time,collected the mouse serum,used flow cytometry to detect same cytokines in serum,and evaluated its immune effct in vivo. Results: Bufadienolides synergistically with Con A can stimulated the lymphocyte secreting INF-γ,GM-CSF,IL-1α,IL-4,IL-5 and IL-6,while singly can stimulate TNF-α,IL-2,IL-4 and IL-5 secretion. With LPS synergistic effect only stimulated TNF-α and IL-6,mostly showed the inhibitory effect on INF-γ,GM-CSF,IL-4 and IL-5. The concentrations of 0. 05 and 10μg/ml are the important groups of leading stimulation and inhibitory. The drug's immune effect on pancreatic cancer tumor nude mice in vivo was observed,compared with the positive control: gemictabine and cinobufotalin injection,bufadienolides can enhance more cytokines expressions. The 1mg/kg bufadienolides had protective effect on spleen while 2mg/kg got the same effect on spleen with gemictabine and cinobufotalin injection. Conclusion: Bufadienolides can promote the proliferation of B cells in a certain extent in vitro,inducing many kinds of cytokines secretion,enhancing the body's immune function,but the mechanism of humoral immunity and the immune effect in vivo needs further research.
【Fund】: 科技部重大新药创制专项(2013ZX09102025)
【CateGory Index】: R285
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