Preliminary Research TLRs/MAPK Signaling Pathway In Role Of Sinomenine Induced Rats ImDC

Background Dendritic cell plays a vital role in transplantation rejection.It may be an important approach to avoiding organ transplant rejection by utilizing immature dendritic cell to induce donor-specific immunologic tolerance.And TLRs and MAPK play a very important role in the maturation of dendritic cells.a large number of studies showed that Sinomenine has immunosuppressive properties.It aims to clarify the action mechanism of Sinomenine in inducing im DC which provides a reliable experimental evidence for applying it in renal transplantation in clinical work.Objective Western blot is analysis for the expression of TLRs,p-ERK1/2,p38 MAPK and p-JNK in Sinomenine induced murine DCs,The study will to investigate the effect of Sinomenine on the differentiation and function of dendritic cells.And understanding of the molecular mechanisms of Sinomenine on immune function,provides the experimental basis for the application of Sinomenine in immune tolerance of organ transplantation.Methods In vitro culture wisatr rat bone marrow-derived precursor cells,observing the morphological differences of DCs were treated with or without Sinomenine under microscope.CD phenotype characteristics of DCs subjected to flow cytometry.with LPS treated for DCs maturation, flow cytometry analysis for LPS-induced DCs TLRs expression with or without Sinomenine.DCs were transfected with or without Sinomenine,then stimulate by LPS,the ERK1/2,p38 MAPK and JNK phosphorylation and non phosphorylation were measured by western blot.Results 1,There is no morphological features of DCs maturation under Sinomenine treatment by Microscope,and DCs phenotype CD86, CD80 were in low expression states.2,The results of flow cytometry indicated SIN + LPS group significantly down-regulate the DCs surface expression of TLR2(84.6±0.34%)、TLR3(83.3±0.14%)、TLR4(83.8±0.16%),Compared to the LPS group of TLR2(94.35±0.16%)、TLR3(97.55±0.16%)、TLR4(94.6±0.12%),The Western blot results showed that SIN significantly inhibit the expression of LPS-induced protein phosphorylation MAPK in DCs,including ERK1/2、p38MAPK、JNK(p<0.05).Conclusions Sinomenine significantly inhibited the expression of TLRs and phosphorylation of ERK1/2, p38 MAPK and JNK in dendritic cells,suggesting Sinomenine suppresse TLRs and MAPK signal pathways and inhibit the DCs of matruation,inducing immune tolerance.