Study On The Biphasic Regulation Mechamism Of IRPS On The Basis Of Transcription Factors T-bet And GATA3

In order to clarify the inner mechanism which the biphasic regulation effects of IRPS, confirm the key factor of the biphasic regulation and provide theory foundation for the two-ways regulation effects of Chinese medicine, we established the immunosuppressive and immuno-hyperfunction rat model with cyclophosphamide. Using those rats models, we further studied the effect that IRPS on different immune status rats from the aspects that transcription factors, cytokines, and the body’s immune at different time points.The five experiments were done.Experiment 1: After rats were feed with different doses of IRPS, we confirmed that the optimal dosage of IRPS affecting rats was 60mg/kg, by measuring the spleen cell proliferation, the activity of NK cell and the content of serum Ig G. And the study made preparations for the whole experimental study.Experiment 2: In order to establish the immune-suppression and immunohyperfunction animal model, rats were injected intraperitoneally with different doses of cyclophosphamide(Cy) in different ways at different time. And then the rats model were evaluated from the aspects of immune organs, immune cells, cytokines and antibody. The results showed that immuno-suppression in rats could be induced by Cy 125mg/kg or 100mg/kg after 6h immunization and immuno-hyperfunction rats could be induced by the method that 20 mg/kg of Cy was injected 1 time before 3d of immunization OVA.Experiment 3: Rats in different immune status were given 60mg/kg IRPS by gavage, lymphocyte proliferation, the activity of NK cell and the content of serum Ig G were detected on 6d and 12 d after immunization. The results showed that IRPS displayed bidirectional regulation on the proliferation of B lymphocyte cells that different immune status and had unobvious effect on the proliferative of T lymphocyte cells in immune-normal and immuno-suppressive rats on the 6d, on the 12 d of immunization, IRPS mainly played a role in immunopotentiation and could enhance the immunological function that immune-normal and immuno-hyperfunction rats. For immuno-suppressive rats, IRPS played to the role of bidirectional regulation, at the same time there were the effects of immunopotentiation.Experiment 4: Rats in different immune status were given 60mg/kg IRPS by gavage, the contents of Th1/Th2 cytokines and inflammatory cytokines which different immune rats were detected at different time point. The results showed that IRPS could relieve decreasing trend of Th1/Th2 cytokines that immuno-hyperfunction rats, have certain inhibitory effect on the increasing trend of Th1/Th2 cytokines that immune-suppression rats and have dual immunomodulatory activity for the secretion of inflammatory cytokines that different immune rats. All these results indicated that IRPS could play an different role on the expression of cytokines depending on different immune status of rats.Experiment 5: Rats in different immune status were given 60mg/kg IRPS by gavage, the expression of m RNA that transcription factor T-bet and GATA3 in different immune rats were detected at different time point. The results showed that IRPS had a significant biphasic regulation effects on the ratio of T-bet/GATA3 that immune-suppression rats, could reduce the ratio of T-bet/GATA3 that immunenormal rats and could also reduce ratio of T-bet/GATA3 that immuno-hyperfunction in advanced stages of testing.The experimental results indicated that IRPS had a biphasic regulation effects on the rats that immuno-suppressive and immuno-hyperfunction rats by affecting the secretion of cytokines and the expression of transcription factors, and could enhance immune function that immune-normal rats.And at the same time, our results were also important reference data for the further study that possible targets of IRPS affecting body.