The effect of recombination adenovirus expressing nuclear factor-κBp65-specific small interference RNA in vivo
Honghong Wang
Abstract
Honghong Wang
Abstract
Objective To observe the inhibitory effect of the recombinant adenovirus expressing nuclear factor-κB (NF-κB)p65-specific small interference RNA (siRNA) in knee joints of rats. Methods The recombinant adenovirus was injected into knee joints of SD rats to optimize the dose of the recombinant adenovirus. The staining of β-gal was used to determine the tissue specificity of the recombinant adenovirus, the effect of recombinant adenovirus on activation of NF-κB was measured with electrophoretic mobility shift assay ( EMSA), and Western blotting was used to measure the effect of the recombinant adenovirus on the expression of NF-κB in chondrocytes and synovial cells. Results The optimal dose of the recombinant adenovirous in SD rat joints was 1.6 × 108 pfu/ml. Staining of β-gal indicated that the recombinant adenovirus could be transfected into the cartilage and synovium. EMSA and Western blotting revealed that the recombinant adenovirus could be expressed in chondrocytes and synovial cells, reduce the activation of NF-κB and inhibit the expression of NF-κBp65. Conclusion The recombinant adenovirus expressing NF-κBp65-specific siRNA can reduce the activation of NF-κB, and inhibit the expression of NF-κBp65 in knee joints of rats. Key words: NF-κB; siRNA; Adenovirus
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Objective To observe the inhibitory effect of the recombinant adenovirus expressing nuclear factor-κB (NF-κB)p65-specific small interference RNA (siRNA) in knee joints of rats. Methods The recombinant adenovirus was injected into knee joints of SD rats to optimize the dose of the recombinant adenovirus. The staining of β-gal was used to determine the tissue specificity of the recombinant adenovirus, the effect of recombinant adenovirus on activation of NF-κB was measured with electrophoretic mobility shift assay ( EMSA), and Western blotting was used to measure the effect of the recombinant adenovirus on the expression of NF-κB in chondrocytes and synovial cells. Results The optimal dose of the recombinant adenovirous in SD rat joints was 1.6 × 108 pfu/ml. Staining of β-gal indicated that the recombinant adenovirus could be transfected into the cartilage and synovium. EMSA and Western blotting revealed that the recombinant adenovirus could be expressed in chondrocytes and synovial cells, reduce the activation of NF-κB and inhibit the expression of NF-κBp65. Conclusion The recombinant adenovirus expressing NF-κBp65-specific siRNA can reduce the activation of NF-κB, and inhibit the expression of NF-κBp65 in knee joints of rats. Key words: NF-κB; siRNA; Adenovirus
Key concepts: Recombinant DNA, Molecular biology, Blot, Chemistry, Adenoviridae, Transfection, Biology, Gene