Screen Out Of Chinese Traditional Medicines On The Effect Of Promoting Axon Elongation And Regeneration To The Rat Retinal Tissue In Three-dimensional Culture System

Chinese traditional medicines have been widely used in the treatment and research in ophthalmology.We noted that the research of Chinese traditional medicines almost focus on the respect of neuroprotection,and has rare systematic investigation and research on neuroregeneration.So we want to know if Chinese traditional medicines can improve the regeneration of retinal ganglion cells axons.We choose four kinds of herb monomer-puerarin,tetramethylpyrazine,bilobalide-B and saffron-I and saffron and using three-dimensional culture system to screen out and compare their effect on the regeneration of retinal ganglion cells axons.Part one:Tetramethylpyrazine promotes retinal ganglion cell axon regeneration in three-dimensional culture system of rat' s retinal tissueObjective:To investigate the effects of tetramethylpyrazine on the axon regeneration of retinal ganglion cells(RGCs) in vitro,different concentrations of tetramethylpyrazine were applied to the three-dimensional culture of rat' s retinal tissue system.Methods:The retinal tissues of SD rats aged 1-3d were cut into pieces of 0.5mm×0.5mm,and cultured in three-dimensional culture system. Investigated the effects to different concentrations of tetramethylpyrazine(0.125g/l,0.25g/l,0.5g/l,1.0g/l) on the growing of RGCs axons,using fluorescence inverted microscopy and immunohistochemical staining.The number and length of outgrowing RGCs axons were calculated in days of 3,6,9. Results:The number and length of regrowing axons to all of the tetramethylpyrazine treated were larger than that in the control,among which the effect to 0.5g/l tetramethylpyrazine was most obvious(p＜0.05). Thy-1.1 and GAP-43 double positive fibers shown to be regenerated axons.Conclusion:Tetramethylpyrazine improves RGC axons regeneration and elongation in three-dimensional culture system at certain concentration.Part two:Screen out and compare the effects of four kinds of herb monomers on the regeneration of retinal ganglion cells axonsObjective:To screen out and compare of herb monomers(puerarin,tetramethylpyrazine,bilobalide-B and saffron-I )on the regeneration of retinal ganglion cells(RGCs) axons in vitro.Methods:The retinal tissues of SD rats aged 1-3d were cut into pieces of 0.5mm×0.5mm,and cultured in three-dimensional culture system. Investigated the effects to different concentrations of puerarin,tetramethylpyrazine,bilobalide-B and saffron-I(0.125g/l,0.25g/1,0.5g/l,1.0g/l) on the growing of RGCs axons,using fluorescence inverted microscopy and immunohistochemical staining.The number and length of outgrowing RGCs axons were calculated in days of 3,6,9.Results:The number and length of regrowing axons to all of the puerarin tetramethylpyrazine,bilobalide-B and saffron-I treated were larger than that in the control,among which the effect to bilobalide-B was most obvious(p＜0.05).Thy-1.1 and GAP-43 double positive fibers shown to be regenerated axons.Conclusion:Puerarin,tetramethylpyrazine,bilobalide-B and saffron-I could improve the regeneration and elongation of cultured RGCs axons,and bilobalide-B is the activest.