Abstract:

Objective This article tests a recombinant plasmid pEGFP -iNOS encoding human inducible nitric oxide synthase （iNOS） transient expression in rabbit periodontal ligament cells, and this may contribute to transfer iNOS gene to animal periodontal tissue in vivo. Methods Rabbit periodontal ligament cells were transfected with pEGFP -iNOS by means of lipofectamine media methods. Transient transfection were evaluated by fluorescent microscope. After transferring 12 h and 48 h, the transcription of iNOS gene was tested by quantitative real-time PCR and the expression product of pEGFP-iNOS was identified by Western blot. Blank plasmid-transfected group and non-transfected group were used as control. Results The expression of green fluorescence protein was detected in 12 h after transferring. The transcription of iNOS mRNA and expression of iNOS protein in cells transfected with pEGFP -iNOS were detected by real -time PCR and Western blot. Conclusion The iNOS gene was transfected successfully, the exogenetic iNOS mRNA can be correctly transcribed and expressed in rabbit periodontal ligament cells. In vivo gene transfer of iNOS to rabbit periodontal tissue is feasible.

Key words: nitric oxide, inducible nitric oxide synthase, rabbit periodontal ligament cells, transient transfection