NGF Activate PC-12 Cells Na+ Ionic Current And The Regulation Of IRF-1

Objective:The expression of NGF is marked heighten in lungs after RSV infected, and the mechanism is not distinct whether NGF enters changes of airway neuroplasticity after RSV infected.To studing NGF regulate the level of Na+ ionic current changes in PC-12 cells of the type sensory neuron, and to exploring the reguration of the IRF-1 and to clarify the airway neuroplasticity mechanism that RSV is infected.METHODS:1.Using different concentrations of NGF(0ng/ml,50ng/ml,200ng/ml) to stimulate PC-12 cells, Then Na+ channel current (INa+) changes were recorded by using whole-cell patch-clamp techniques. 2.Using different concentrations of NGF (0ng/ml～200ng/ml) to stimulate PC-12 cells with different hours,and extracte mRNA to do IRF-1 Real-time PCR to detect the IRF-1mRNA expression.3.There were four groups:NGF group, NGF+IFN-ygroup, IFN-ygroup and control group. And after stimulated PC-12 cells 2h, then we extracted nucleoprote to do IRF-1 Western-bloting to detect the IRF-1 expression in nucleus.4.There were three groups to interferring PC-12 cells:control group, negative control group and the experimental group.Using 200ng/ml NGF stimulated PC-12 cells 2h that were interferring, and we recorded the Na+ channel current(INa+) and INa+I-V curve.RESULTS:1.There was a certain concentration-dependent between NGF and Na+ ionic current, when NGF is stimulating PC-12 cells. 2.The highest expression of IRF-1 mRNA is 2h when NGF stimulated PC-12 cells.3.The expression of IRF-1 nuclear protein was remarked when NGF stimulated PC-12 cells.4.When IRF-1 has been disturbaced, current density-voltage relationship curve shifted, Peak of current density of Na+ channel current (INa+) in the experimental group was significantly reduced(12.92±0.55pA/pF) compared to the control group (31.26±1.72pA/pF)and the negative control group (34.81±0.64pA/pF, P<0.05.Compared with the control group and the negative control group, the experimental groupⅠNa+Ⅰ-V curve was down, but Na+ channel voltage-dependent change of activation potential,peak potential,and the I-V curve shape had no effect on trak.Conclusion:1.NGF activated the Na+ channel current (INa+) in PC-12 cells.2.IRF-1 is one of the signal mechanisms to regulating and conrolling the Na+ channel current.