The Expression Of Substance P And NFL In Dorsal Root Ganglion Sensory Neuron Cell Of Rats Was Influenced By MEF2C

Objective: To establish a feasible method of culture and purification of embryonic dorsal root ganglion neurons of rats, To observe the expression of Substance P and NFL in dorsal root ganglion cell influenced by MEF2 and its relationship with airway hyperresponsiveness.METHODS : Adequancy dorsal root ganglions(DRGs) were obtained by microdissection, then were dissociated in trypsin plus EDTA. The dissociated neurons were cultured with NB median and 5-Fluoro-2'-deoxyuridine/Uridine for purification. Anti-NF200 and DAPI staining was to examine the purification rate of DRGn. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF (10 ng/mL, 30 ng/mL, 100 ng/mL, or 200ng/ml) for 24 h. The neurons cultured in media with the lowest concentration of NGF(10 ng/mL) served as control. Real time PCR were used for detecting the mRNAs of Substance P and Nfl in the DRGn cells. transfected three Mef2c-siRNAs into PC 12 cells with the way of chemical transfection respectively, then chosen then best siRNA with highest transfectional rate with Real time PCR. And knocked out gene MEF2C of DRGn cells with chemical transfection, then detecteed then expression of Substance P and Nfl in DRGn cells with Real time PCR after high concentration NGF stimulation.RESULTS: 1.The isolated and cultured DRGn survived and grew nicely in vitro, and the purified rate of DRGn was above 96%. 2. Substance P and Nfl expression increased in primary cultured DRG neurons of rats in a dose-dependent manner of NGF.3.The expression of Mef2c in experimental group decreased 50% than control group, but there was no change of Creb; The Substance P decreased 41% in experimental group than control group at the same time the Nfl was decreased 61%.Conclusion: 1 .The primer culture method is very utility for obtain massive highly purified embryonic dorsal root ganglion neuron cells of rats.2. NGF may promote Substance P and NFL synthesis in rat DRGn cells.3.MEF2C regulated the expression of Substance P and NFL in DRGn cells of rat embryo and MEF2C maybe a critical transcription factor involved in airway hyperresponsiveness.