The Effects Of Nuclear Factor-Kappa B Antisense Oligdeoxynucleotides On Expression Of Interleukin 8 In Peripheral Blood Mononuclear Cells From Patients With Rheumatoid Arthritis

Rheumatoid arthritis(RA) is a complex autoimmune disease .It ischaracterized by the activation of peripheral blood lymphocytes, chronicinflammation in the joints, progressive destruction of cartilage and bone ,functional disability. NF- B play a key role in the pathogenesis of RA.Among the genes regulated by NF- B are a number of cytokines,chemokines, adhesion molecules and proteases. Nuclear factor-kappa Bantisense oligdeoxynucleotides is a new way to inhabit activity of nuclearfactor-kappaB. Inhibition of NF- B can be a novel target to treatmentRA,and a new way to screen anti-rheumatic drugs. This study was designed to illuminate the expression of interleukin-8 (IL-8) in peripheral blood mononuclear cells (PBMC) from RApatients , and the effects of nuclear factor-kappaB antisenseoligdeoxynucleotides on expression of IL-8 in PBMC from patients withRA,explain the molecular anti- rheumatic mechanism of nuclear factor-kappaB antisense oligdeoxynucleotides. Objective To investigate Nuclear factor-kappaB p 65 antisenseoligdeoxynucleotides (AS ODN) might affect the expression of NF- Bp65 and IL-8 in peripheral blood mononuclear cells (PBMC) from patientswith Rheumatoid Arthritis and explain the molecular anti- rheumaticmechanism of AS ODN of Nuclear factor-kappaB p 65. Method PBMC samples were obtained from 10 patients with RAas defined by the criteria of the American College of Rheumatology .PBMC was isolated , and cultured in RPMI-1640 with and without NF- Bp65 antisense oligdeoxynucleotides ( AS ODN), Misssenseoligdeoxynucleotides (MS ODN) and sense oligdeoxynucleotides (SODN ) . NF- B p65 was determined by western blot analysis . Theexpression of IL-8 mRNA was studied by reverse transcriptionpolymerase chain reaction(RT-PCR).Levels of IL-8 were measured byenzyme-linked immunosorbant assay ( ELISA). Result 1.NF- B p65 AS ODN inhibited NF- B p65 expression innucleus; 2.PBMC from patient with RA which was incubated in mediumalone may express L-8 mRNA and secret chemokine IL-8; 3 . TNFα stimulated up-regulated PBMC from patient with RAchemokine IL-8 secretion , P<0.05; 4.NF- B p65 AS ODN inhibited TNFα -induced PBMC frompatient with RA which expressed IL-8 mRNA and secreted chemokine IL-8, as compared with that treated PBMC from patient with RA which wasincubated in medium alone , TNFα -induced PBMC from patient with RA ,NF- B p65 S ODN and NF- B p65 MS ODN. NF- B p65 AS ODNresulted in down-regulation of IL-8 mRNA and strikingly reduction of IL-8levels, P<0.01;. Conclusions 1.PBMC from patient with RA which was incubated in mediumalone expressed IL-8 mRNA and secreted chemokine IL-8; 2.TNFα up-regulated PBMC from patient with RA IL-8 secretion ; 3.NF- B p65 AS ODN strikingly inhibited TNFα -induced PBMCfrom patient with RA which expressed and secreted IL-8.