Efficacy of mIL-18BP and mIL-4 adenoviral coexpression gene therapy for cyclooxygenase-2 and inducible nitric oxide synthetase in murine collagen-induced arthritis / 中华风湿病学杂志

ABSTRACT

Objective A recombinant adenoviral vector containing mIL-18BP and mIL-4 fusion gene(AdmIL-18BP/mIL-4) was constructed and used to investigate the role of mIL-18BP and mIL-4 in medula-ring the expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthetase (iNOS) and their inducing products PGE2, NO in murine collagen-induced arthritis. Methods Male DBA-1/BOM mice were used in this study. Mice with CIA were intra-articularly injected with 107 pfu/6μl of AdmIL-18BP/mIL-4.Intra-articular injections of AdLacZ or PBS were used as controls. The mRNA expression of COX-2, iNOS in synovial tissue was analyzed by semi-quantitative RT-PCR. Expression of COX-2 and iNOS protein was estimated by Western blot method. The production of PGE2 and NO in synovia was detected by competitive ELISA and enzyme reduction of nitrate. Results The expression of COX-2, iNOS mRNA in routine synovial tissue of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group (0.15 vs 0.42,P<0.01 ; 0.05 vs 0.77, P<0.01) and PBS group (0.15 vs 0.65, P<0.01; 0.05 vs 0.64, P<0.01 ). And the protein expression of COX-2, iNOS from AdmIL-18BP/mIL-4 treatment group was also obviously lower than that of AdLacZ group (0.08 vs 0.92, P<0.01; 0.11 vs 1.00, P<0.01) and PBS group (0.08 vs 0.77, P<0.01; 0.11 vs 0.84, P<0.01 ). The PGE2 and NO production in synovia of AdmIL-18BP/mIL-4 treatment group was significantly lower than that of AdLacZ group [(0.68x0.06) vs (2.58±0.21)ng/mL, P<0.01; (23.4+2.5) vs (60.0±11.3)μmol/L, P<0.01 ] and PBS group [(0.68±0.06) vs (2.57±0.20)ng/mL, P<0.01; (23.4+2.5) vs (60.3±13.4)μmol/L, P<0.01]. Conclusion These data indicat that local over-expre-ssion of mIL-18BP and mIL-4 can down-regulate COX-2, iNOS and their induced product PGE2, NO in CIA mice. The combination treatment with mIL-18BP and mIL-4 is a promising therapeutic target for RA.