Effect Of Sinomenine On TLR2/TLR4-MyD88/TRIF Signaling Pathway In DC2.4 Cells

Objective:Study on the effect of sinomenine on TLR2 and TLR4 receptor of DC cells and the influence of MyD88 dependence signal pathway or MyD88 non-dependence signal pathway,and then clear the inhibition mechanism of sinomenine on DC cell activation.Thus infer sinomenine in the treatment of rheumatoid arthritis,the key point in the process of development of new drugs for the treatment of rheumatoid arthritis and to provide evidence of pharmacology and medical aspects.Methods:1,using Western Blot method to detect DC2.4 cell's the expression of TLR2 and TLR4,MyD88and TRIF;2,real-time qPCR fluorescence quantitative method to detect DC2.4 cell's expression of TLR2 m RNA,TLR4 mRNA,MyD88 mRNA and TRIF mRNA.Results:1,through the experiment,we found that the expression of TLR2 and TLR4 and MyD88 and TRIF protein were significantly increased in DC2.4 cell model group compared with the blank group,which were stimulated by LPS,while the related proteins expression in sinomenine group have varying degrees of decline after the intervention of drug;2,LPS can increasese the expression of proteins and corresponding mRNA about TLR2 and TLR4,MyD88 and TRIF in DC2.4 cell.While the related mRNA decreasess in inomenine drug treatment group.Such as the model group is 3.50 times than the blank group about the 2^-?35??35?CtCt value of TLR2mRNA expression.While the sinomenine drug treatment group was 1.40 times than the blank control group,the model group is 1.47 times than the blank group about the TLR4 mRNA expression,the sinomenine drug treatment group was 0.90 times than the blank control group.About the 2^-?35??35?CtCt value of MyD88 mRNA expression quantity,the model group is 2.41 times the number of that blank group,the number of the sinomenine drug group MyD88 m RNA is 1.08 times that of the blank group.About the 2^-?35??35?CtCt value of TRIF mRNA expression quantity,the model group is 6.67 times up on the blank group's value,while the sinomenine drug group is 1.43 times that of the blank group.Conclusion:The experimental result indicates:TLR2 and TLR4 in the pathogenesis of RA may be stimulated by specific antigen at the same time,then activating the downstream's two kinds of MyD88 dependence's and MyD88 not dependence's signaling pathways,thus activating the antigen presenting cells-DC.The abnormal activation of DC cell and the abnormal activation of T cells are the core of the development of RA disease pathogenesis.And the sinomenine could inhibit TLR2/TLR4-MyD88/TRIF signaling pathways,and inhibite DC cell's activation and play a pharmacological effects for the treatment of RA.