The purpose of our study was to explore the regeneration effect of the ultrasound (US)-targeted microbubble destruction-mediated eukaryotic coexpression vector (pIRES2-enhanced green fluorescent protein [EGFP]/hepatocyte growth factor [HGF]) with EGFP and HGF gene system on facial nerve injury in rats. Forty rats were randomly divided into 4 groups after the models of facial nerve injury were established: A, phosphate-buffered saline (PBS) group; B, HGF and microbubble (HGF + MB) group; C, HGF and US (HGF + US) group; and D, HGF + US + microbubble (HGF + MB/US) group. Gene and protein levels of HGF were detected by quantitative real-time reverse transcriptase-polymerase chain reaction and Western blot, respectively. The expression of pEGFP in facial nerve trunks was examined by laser scanning confocal microscope; HGF gene and protein expression were significantly higher in D group compared with those of the other groups (P < 0.05). The expression of pEGFP was the strongest in D group (P < 0.05). These data indicate that US-targeted microbubble destruction effectively transfects the HGF gene into target tissues and has a significant effect on an injured facial nerve, thus providing a new strategy for gene therapy in facial nerve injury.