Low dose of ouabain protects injury of spiral ganglion neurons in vitro / 临床耳鼻咽喉头颈外科杂志

ABSTRACT

OBJECTIVE@#To evaluate the protective effect of low dose ouabain on injury of cultured spiral ganglion neurons evoked by trophic factors deprived and to explore the mechanism.@*METHOD@#Spiral ganglion neurons were cultured in vitro for 7 days, and then exposed to Neurobasal medium + B27 supplement, Neurobasal medium only or Neurobasal medium + 10 nmol/L ouabain, respectively. After 48 h exposed to different medium, spiral ganglion neurons were stained by FITC labeled Annexin-V and PI, then apoptosis index were calculated using fluorescent microscope and flow cytometry, respectively. In addition, spiral ganglion tissues were cultured for 48 h to evaluate dendrite growth of spiral ganglion neurons in each group. Immunocytochemistry were performed to detect the level of Bcl-2 in each group at 6 h and 12 h.@*RESULT@#Spiral ganglion neurons exposed to Neurobasal medium +10 nmol/L ouabain have a similar apoptosis index compare with that of Neurobasal medium + B27 supplement, but a much lower apoptosis index than that of Neurobasal medium only. In addition, dendrite growth of spiral ganglion neurons exposed to Neurobasal medium +10 nmol/L ouabain was much longer than that of Neurobasal medium only. Bcl-2 level increased in spiral ganglion neurons exposed to Neurobasal medium + 10 nmol/L ouabain at 6 h.@*CONCLUSION@#Low dose of ouabain protects injury of spiral ganglion neurons evoked by trophic factors deprived in vitro. This effect may mediated by increasing the level of Bcl-2.