Objective: We test a recombinant plasmid pEGFP-iNOS encoding human inducible nitric oxide synthase(iNOS) transient expression in rabbit periodontal ligament cells,this may contribute to transfer iNOS gene to animal periodontal tissue in vitro.
Methods: Rabbit periodontal ligament cells were transfected with pEGFP-iNOS by means of lipofectamine media methods. Transient transfection were evaluated by fluorescent microscopy.After transfected 12hours and 48hours, the transcription of iNOS gene was tested by quantitative real-time PCR and the expression product of pEGFP-iNOS was identified by Western-blotting.We used blank plasmid-transfected group and non-transfected group as controls.
Results: The expression of green fluorescence protein was detected after transfected 12hours.The transcription of iNOS mRNA and expression of iNOS protein in cells transfected with pEGFP-iNOS were detected by real-time PCR and Western-blotting.
Conclusions: 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.