Construction Of DJ-1 Gene Lentivirus And Transfection Of Cochlear Spiral Ganglion Cells

Objectives:The animal model for Spiral Ganglion Cells(SGCs)with varying degrees of damage was established by damaging Spiral Ganglion Cells(SGCs)with different concentrations of Ouabain on the round window membrane of Mongolian gerbils.The dj-1 gene was successfully packaged with lentivirus vector,and the lentivirus transfection in SGCs cells were observed,and the protective effect of dj-1 on SGCs injury was determined.Methods:①Male Mongolian gerbils with normal hearing were randomly divided into experimental group and control group.Artificial perilymphatic fluid(APF)was drip onto the round window membrane of the right ear of gerbils in control group,and a total of 0.2ml Ouabain with a concentration of 33 μ M,66 μ M and 80 μ M was drip onto the round window membrane of the right ear of gerbils in experimental group to establish the animal model.②The dj-1 gene was successfully packaged with lentivirus vector.③Male Mongolian gerbils with normal hearing were randomly divided into experimental group and control group.Artificial perilymphatic fluid(APF)and lentiviru with DJ-1 was drip onto the round window membrane of the right ear of gerbils in control group,and a total of 0.2ml Ouabain with a concentration of 33 μM,66 μM,80 μ M and lentiviru with DJ-1 was drip onto the round window membrane of the right ear of gerbils in experimental group.The protein expression was detected by Western Blot two weeks later.The effect of dj-1 on cochlear spiral ganglion cell injury was observed on the basis of protein expression in groups E.Results:Morphological observation showed that the number of SGCs decreased with the Ouabain concentration increasing and began to see swelling cells,cytoplasm were filled with vacuoles,condensed and fragmented nucleus.Compared with the Ouabain group,morphological observation showed that the number of spiral ganglion cells decreased in the dj-1 lentivirus group.Differences associated with P values of<0.05 were considered as the difference was statistically significant.Conclusions:①The animal model for Spiral Ganglion Cells(SGCs)with varying degrees of damage was successfully established.②Dj-1 gene lentivirus can be successfully packaged by 293T cells.③The lentiviral expression vector of DJ-lgene was successfully transfected into SGCs and expressed the corresponding protein.④In our research,dj-1 may has no protective effect in the animal model of cochlea spiral ganglion cells damaged by Ouabain.