Abstract
Objective To construct lentiviral vectors targeting rat interleukin-1 receptor type I（IL- 1 RI） gene by RNA interference （RNAi） and to study its effects on chondrocytes in vitro. Methods Effec- tire sequence of siRNA targeting IL-1RI gene was designed and determined and 239T cells were were trans- fected with pGCL-GFP vector containing the inserted DNA sequence. The lentivirus containing the effective sequence of siRNA targeting IL-1RI gene with concentration of 1 × 107 TU/ml was obtained after supernatant of 293T cells were collected and concentrated. The recombinant lentivirus was used to infect IL-l-stimulated rat chondrocytes previously with multiple of infection （MOI） 20 and 50 seperately in vitro and the expression of IL-1RI and matrix metalloproteinase （MMP） -1 was detected by using Western blotting 3 days later. Re- suits The best MOI for lentivirus infecting chondrocytes was 50. Lentiviruses demonstrated a high （above 70% ） infection efficiency of chondrocytes. The expression of IL-1RI and MMP-1 was both repressed signifi- cantly in rat chondrocytes by RNAi. Conclusion The recombinant lentivirus shows significantly inhibitory effects on IL-1RIand MMP-1 （the downstream protein） in rat chondrocytes. Key words: Lentivirus ; Interleukin-1 ; RNA interference ; Chondrocyte ; Matrix metallo- proteinase-1
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