Preventive effect of plasmid-mediated sphingosine kinase1 gene transfer on intestinal adhesion after operation

Objective To observe the expression of plasmid sphingosine kinase1（SPK1） in a rat model of intestinal adhesion and the protective effect of SPK1 gene transfer on intestinal adhesion in rats after operation. Methods Mesothelial cells isolated from rats were cultured and transfected with pcDNA3-SPK1.Expression of SPK1 protein was detected by Western blot and activity of SPK1 was assayed by ATP incorporation.Transfer efficiency of plasmid vector in mesothelial cells was evaluated using green fluorescent protein（GFP） as a marker.A rat model of cecum brush burn-induced adhesion was established to evaluate the preventive effect of plasmid-mediated SPK1 gene transfer on intestinal adhesion after operation. Results Plasmid pcDNA3-SPK1 effectively transfected the damaged mesothelium and mediated the SPK1 gene expression.Plasmid pcDNA3-SPK1 gene transfer decreased the intestinal adhesion from 80% to 30% in the rat model of intestinal adhesion after operation. Conclusion Plasmid SPK1 can effectively prevent intestinal adhesion in rats after operation.