Abstract:

BACKGROUND: Recently, interleukin-17 (IL-17) is proved to be a pro-inflammatory cytokine, which produced by CD4+helper T cells subtype Th 17 cells. IL-17 is expressed in T cells, and IL-17 receptor is expressed in subtotal cells and tissues.
OBJECTIVE: To explore the synergetic effects of IL-17 binding IL-17 receptor (IL-17RA, IL-17RC) and tumor necrosis factor α on expression of inducible nitric oxide synthase (iNOS) promoter in the synovium as well as synthesis of nitric oxide.
METHODS: mRNA of human knee joint cartilage and synovial tissue was extracted. RNA interference expression vector of IL-17RA and IL-17RC were constructed together, and transfected into 293T cells. IL-17 receptor-deficient expression cell model was established under the G418 selection pressure. The recombinant eukaryotic plasmids containing iNOS promoter were transfected into wild type 293T cells and 293T cells deficient in IL-17RA expression respectively. Through the synergetic induction of IL-17 and tumor necrosis factor α, expression of iNOS promoter and activity of iNOS were observed by detection of luciferase and nitric oxide concentrations.
RESULTS AND CONCLUSION: ①Full-length cDNA from human knee joint cartilage and synovial tissue iNOS promoter and eukaryotic expression plasmids pGL3-iNOS-p luciferase were obtained successfully. ②Cell model of IL-17RA expression deficiency was established. ③In IL-17RA-deficient expression cells, the expression of iNOS promoter and activity of iNOS were significantly reduced. ④IL-17RC specific shRNA eukaryotic expression vector of shRNA-IL-17RC was successfully constructed. These findings suggest that IL-17 via its receptor IL-17RC cooperated with iNOS promoter to obviously promote gene expression of iNOS and nitric oxide production.