Impact of lentivirus-mediated siRNA interfering MyD88 on rat alveolar macrophage

Objective To detecte rat alveolar macrophage function after using RNA interference technology,to interfere expression of MyD88 on rat alveolar macrophage by constructing recombinant lentivirus that express rat myeloid differentiation factor 88(MyD88) siRNA.Methods Designed and constructed 3 plasmid expressing MyD88 siRNA,then respectively co-transfected them into HEK- 293T cells with plasmid expressing MyD88.Western blotting detected the expression of MyD88 in order to filter the most effective siRNA plasmid.Constructed recombinant lentivirus by Gateway and determined the titer,then transfected them into endotoxin-stimulated rat alveolar macrophage and the uninfected cells were used as control group,finally determined concentration of IL-1βand IL-6. Results Successfully filtered the most effective siRNA plasmid and constructed recombinant lentivirus. The titer of recombinant lentivirus was 2.0×10~6 TU/ml.IL-1βand IL-6 of NR8383 cell were significantly reduced after infecting recombinant lentivirus compared with control group.The difference has statistical significance.Conclusion Lentivirus-mediated RNAi can effectively suppress MyD88 expression and significantly reduce the release of cytokine in rat alveolar macrophage,which may provide a possibility that treat rat lung transplantation-related obliterative bronchitis(OB) by targeting antigen-presenting cells(APC) in vivo.