Study Of The Immunosuppressive Effect Of Gossypin On Mouse Th1/Th2 Cells

The immune system is an important disease defense system in an organism.Normally,the immune system detects pathogens entering the body and can distinguish pathogens from normal tissue.The body’s high immunity can lead to"anaphylactic"reactions and autoimmune diseases."Anaphylactic"reaction refers to the allergen after contact with the body,causing strong irritation,runny nose,sneezing,itching and other symptoms.Many common substances in life can become allergens,causing the body to produce"allergic"reactions,such as pollen,catkins,animal dander and other things can become allergens.Autoimmune diseases are a class of diseases caused by the autoimmune system mistakenly attacking its own cells.Common autoimmune diseases include pernicious anemia,atrophic gastritis,autoimmune glomerulonephritis and others.Immunosuppressants are a class of chemical or biological substances that reduce the degree of tissue damage by inhibiting the body’s production of excessive immunity.At present,immunosuppressants are used clinically for a wide range of diseases,including rejection after organ transplantation and the treatment of autoimmune diseases such as dermatomyositis,scleroderma,multiple sclerosis,myasthenia gravis,demyelinating disease,and primary adrenal cortex atrophy.Drugs such as prednisone(PDN),cyclophosphamide(CTX)and methotrexate(MTX)have been used in clinical practice,but the toxicity of immunosuppressive drugs has become one of the obstacles to the clinical treatment of related diseases,so it is urgent to find an immunosuppressive drug with few toxic side effects and a wide range of sources.China’s natural plant sources are very rich,and a large number of studies have shown that drugs derived from natural plants can act on immune cells and play a role in suppressing or promoting the immune system,thereby playing an effect on the regulation of the immune system.In this study,a traditional Chinese medicine monomer with potential immunosuppressive function,Gossypin,was screened through preliminary screening.The specific research content is as follows:the effective dose range of Gossypin on mouse splenic lymphocytes was screened by CCK8 experiment,and the appropriate concentration was selected for mouse splenic lymphocytes proliferation experiment.The inhibitory effect of Gossypin on related cytokines secreted by Th1 and Th2 cells was detected by ELISA,and the inhibitory effect of Gossypin on cytokine mRNA was detected by RT-PCR experiments.Flow cytometry detected the effect of Gossypin on the distribution of ConA-induced T cell subtypes CD4~+T cells and CD8~+T cell subsets.Western Blot investigated the inhibitory effect of Gossypin on JAK-STAT and NF-κB pathway proteins.By establishing delayed-type hypersensitivity(DTH)mouse model and ovalbumin(OVA)mouse model,the swelling degree of the auricle,the pathological tissue section of the auricle of the mice were detected,the thymus and spleen index,cell proliferation,cytokines and the secretion changes of related antibodies were detected,and the inhibitory effect of Gossypin on cellular immunity and humoral immunity response was verified.The results showed that Gossypin could inhibit the proliferation of related splenic lymphocytes,and its inhibition effect was dose-dependent.It can inhibit the levels of cytokines IL-2,IFN-γsecreted by Th1 cells and cytokines IL-4 and IL-6 secreted by Th2 cells,and its inhibitory effect is by inhibiting the synthesis of cytokine mRNA.To a certain extent,Gossypin can inhibit CD4~+and CD8~+T cells subsets in T cells at experimental concentrations,have a significantly inhibitory effect on CD4~+/CD8~+ratio,and inhibit the activation of JAK-STAT and NF-κB pathway-related proteins.After adding Gossypin to the DTH model,the degree of swelling of the ear pinna,the degree of auricle tissue lesions and the thymus and spleen index of mice were gradually improved,and the levels of cytokines IL-2,IFN-γ,IL-4 and IL-6 secreted by Th1 and Th2 cells were significantly inhibited.The addition of Gossypin in the OVA mouse model significantly inhibited the production of IgG antibody and IgG antibody subtypes IgG1 and IgG2a,as well as the secretion of related cytokines IL-2,IFN-γ,IL-4 and IL-6.In summary,the effect of Gossypin on Th1 and Th2 cells in mice was comprehensively studied in vitro and in vivo,which can reduce the number of splenic lymphocytes and the secretion of Th1and Th2 cytokines,inhibit the activation of JAK-STAT and NF-κB pathways,and prevent related protein synthesis,indicating that Gossypin can act on cellular immunity and humoral immunity at the same time to exert immunosuppressive effects.