Effects Of Methylprednisolone On Neurotrophic Factor Secretion Of Schwann Cells In Vitro

Objective:To research the effects of Methylprednisolone on Schwann cells in vitro of secreting the neurotrophic factor.Methods:Chose 6 New Zealand rabbits of 2 weeks-old, killed and soak in ethyl alocohol of 75% for 20 minutes. Procurexing the ischiadic nerve and brachial plexus of both sides under asepsis, striping the epineurium and cutting to be chips of 1 mm3, and then grow in the culture flask which spreading by gelatine with 1 cm distance. Passage after 7days, go on cultivanting 3 days, then trypsinizating and depuration by differential adhesion way. Accredit the purity of cells with S-100 protein-immunohisto-chemistry stain, and preparing to be cell suspension of 5×10⁴ /ml, load into 4-hole cultivation board with 1 ml/hole.According to the concentration of the drug, there was divided into four groups: A group, control group, only giving DMEM- culture fluid 2ml; B group, adding 0.1mg / L methylprednisolone into the culture fluid; C group, adding 1.0mg/L methylprednisolone into the culture fluid; D group, to join 5.0mg/L methylprednisolone into the culture fluid. Cultivate in the homeothermia incubator of 37℃with 20% CO₂. Observe the cell growth and estimate the cell number. Every two days, draw-off the culture fluid and keep in the refrigerator of 4℃, wash the board by PBS, then join the new culture fluid with different group.Calculate the increase value of the cell number, get the quantitative assay of NGF,CNTF,GDNF by ELISA, and Calculate the increase value. The statistics test was carried out with one-side variance analysis by SPSS 13.0, choosing statistical significance of P<0.05.Results:①B, C, D groups Schwann cells faster than the growth rate of Group A, three kinds of neurotrophic factor secreted significantly more than group A, statistically significant differences (P <0.01), B, C, D Group Between the secretion was no significant difference between the number of (P> 0.05);②NGF of the test results are better than that of CNTF, GDNF trend (P <0.01), CNTF and GDNF promote the secretion of the situation there was no significant difference (P> 0.05).Conclusion:Methylprednisolone can promote the proliferation of Schwann cells in vitro and the secretion of neurotrophic factor. Application of low-dose and high-dose has no significant differences.Methylprednisolone can promote the regenerate and repair of peripheral nerve by promoting the proliferation of Schwann cells and the secretion of neurotrophic factor.