Nourishing Piyin Remedy On The Protective Effect Of Dorsal Root Ganglion Neurons Under Hypoxic

Objective:To study the protective effect of Nourishing Piyin Remedy on dorsal root ganglion neurons under hypoxic injury and to explore its mechanism.Methods:1. dorsal root ganglion (DRG) was extracted from 24h newborn SD rat, and DRG neurons were cultured adherently. And cells were purificated by adherent differentiation and Ara-C.2. DRG neurons were identified through immunofluorescence four days after purification. Cells under 24-hour preprotection were examined through immuno fluorescence in order to compare the number of cells between each group and observe the outgrowth of processes.3. The rate of cell preservation and viability were assessed by MTT , figuring out the most suitable drug concentration.4. Use infrared spectra to determind components of drug-containing serum and blank serum in an intend to confirm Nourishing Piyin Remedy was in drug- containing serum .5. Use Na2S2O4 to establish hypoxic injury model.The level of SOD in the cell culture supernatant was determined by enzymatic xanthine oxidase essay.Level of MDA in cell culture supernatant was determined by TAB essay.Level of LDH was Determined by enzyme viability essay.6. Statistical analysis: All tests were repeated 10 times,datas were analyzed by statistical software packages SPSS16.0. The results of data using mean±standard deviation of that group differences between single- sample t test of variance, P <0.05 indicated statistical difference.Results:1. Cell population in drug-containing serum group were significantly greater than that of blank serum group and blank control group under fluorescence microscopy. The outgrowth of processes in drug-containing serum group is more intensive, mesh-like, with a stronger fluorescent brightness.2. After hypoxic injury, cell survival rate in 1% drug-containing serum group was 0.537±0.010, higher than that in the control group of 0.221±0.033 (P <0.01), as well as blank serum group of 0.228±0.039 (P <0.01), and close to the survival rate in NGF group of 0.549±0.006 (P> 0.05).3. 1% drug-containing serum hypoxic injury can prevent SOD from decreasing and reduce the generation of MDA and LDH in DRG neurons. The blank serum with same concentration had no significant pre-protection and anti-hypoxic effect on DRG neurons.Conclusion:1. Nourishing Piyin Remedy -containing serum promotes growth of neurons.2. Nourishing Piyin Remedy -containing serum increases the capacity of anti-hypoxic damage, prevents SOD decreases, and reduces the generation of MDA and LDH ,therefore promotes survival and survival status in DRG neurons under hypoxic conditions .3. 1% drug-containing serum can best promote growth and protective effect without obvious toxicity under anti-hypoxia.