Study On The Inhibition Of Rat Dendritic Cell Maturation By JAK/STAT ANDNF-κB Signaling Pathway In Sinomenine

Background:In recent years,with the development of surgical organ transplantation technology and gradually mature,allogeneic kidney transplantation has become the current treatment of end-stage renal disease(renal failure)the most effective means.The factors that determine the success of renal transplantation is not only the success of surgery,more importantly,whether the occurrence of acute and chronic rejection.Sinomenine is mainly used for clinical treatment of various rheumatism and arrhythmia are familiar,and in recent years with its in-depth study,found that it also has anti-inflammatory and immune suppression Pharmacological effects.Therefore,to explore the role of sinomenine in the field of transplantation immunology will help us to lay the foundation for the clinical treatment of organ transplant rejection.Purpose:By detecting the activity or expression level of NF-κB、 JAK and STAT in dendritic cells derived from rat-derived rats.To investigate the effects of sinomenine on the differentiation,maturation and function of rat-derived dendritic cells through JAK / STAT and NF-κB signaling pathways,and to study its mechanism of immunosuppressive for its clinical treatment of organ transplant rejection Provide laboratory basis.Method:1、The rat bone marrow-derived precursor cells were cultured and isolated to obtain im DC(divided into conventional induction group and sinomenine induced group);2、 The phenotype of im DC cells was identified: A.Morphological observation of DC cells under microscope B.The phenotype of DC was analyzed by flow cytometry;3、The expression of NF-κB in DC nuclei was measured by immunohistochemistry(PS staining)(im DC group、im DC/SIN group、im DC/LPS group、im DC/LPS+SIN group);4、The changes of DC DNA and NF-κB binding activity were measured by electrophoretic mobility shift assay(EMSA)(im DC group、im DC/SIN group、im DC/LPS group、im DC/LPS+SIN group);5 、 Western blot was used to detect the expression of JAK and STAT protein(im DC group、im DC/SIN group、im DC/LPS group、im DC/LPS+SIN group).Results:1、Characteristics of DC after sinomenine treatment:(1)The morphology of dendritic cells under microscope was not characteristic of mature dendritic cells;(2)The low expression of CD80 and CD86 in dendritic cells were detected by flow cytometry;The expression of CD80 and CD86 in the sinomenine group was lower than that in the normal induction group,and the diffeence was statistically significant(p <0.05);2 、 Immunohistochemistry NF-κB was expressed in cytoplasm and nucleus of im DC,but the expression of NF-κB was weaker than that of other groups,and the diffeence was statistically significant(p <0.05);3、The activity of NF-κB in the sinomenine group was significantly lower than that in other groups;4、The expression of JAK1,STAT3 and p STAT3 in DCs of SIN group was significantly lower than that in other groups,and the diffeence was statistically significant(p <0.05)。Conclusion : Sinomenine can inhibit DC maturation,In the course of its action,the expression of JAK1、STAT3、p STAT3 protein,the expression of NF-κB and the activity of NF-κB decreased,It was suggested that sinomenine could inhibit the maturation of DC cells by inhibiting JAK / STAT and NF-κB signaling pathway.