The Role Of Peoniflorin On Organotypic Brain Slices Derived From Rat Substantia Nigra Treated With 1-methyl-4-phenylpytidinium In Vitro

[Objective] 1. To investigate the cultural methods of the rat organotypic brain slices derived from substantia nigra. 2. To investigate the methods of making the brain slices treated with 1-methyl-4-phenylpytidinium (MPP+) from the rat with Parkinson's disease. 3. To assess the effects of peoniflorin on dopamine neurons in the brain slices of substantia nigra treated with MPP+.[Methods] 1.The organotypic brain slices culture from substantia nigra (350μm) was prepared from SD neonate rats. The technique of brain slice culture in vitro was carried out on Millicell-CM porous membranes. The HE staining with stereological method, tyrosine hydroxylase (TH) immunohistochemical staining and the cell counting were used to compare with the normal rats'brain slices. 2.Select the brain slices which had been incubated in MPP+ of different concentration for 48 hours, observe the differences among the dopamine neurons from these brain slices. 3.Select the brain slices co-incubated in MPP+ and peoniflorin for 48 hours, observe the effects of peoniflorin on dopamine neurons treated with MPP+.[Results] 1.The substantia nigra cells cultured from 0d to 14d were smaller and with lower density than those of the same age in vivo. TH+ cells cultured were more immature than that in vivo, the occurrence of cell junctions was later, and there were significant difference in the number of TH+ cells(P<0.01) between the slices matured in vitro and in vivo. 2.With the unsharpness edge, the cellular structure of the organotypic brain slices treated with MPP+ was fuzziness, and the cellular density in the slices was lower than that of the slices cultured in vitro homeochronous (the normal control). TH+ cells were sparser than that in the normal control and the cell junctions were decreasing, which was especially conspicuous in the groups treated with mid or high-concentration of MPP+. There was statistical significance of TH+ cell counting between the groups treated with different concentrantions of MPP+ and the control (P<0.05). 3.The cellular structure was vagued in the organotypic brain slices of substantia nigra co-cultured with both peoniflorin and MPP+. While the dead cell masses at the edge were less than those in the groups only treated with MPP+ (control groups). The density of the cells was higher than that in the group treated with mid-concentration MPP+. The cellular structure was clearer with high concentration peoniflorin than that with low concentration. There were more TH+ cell junctions in the group treated with both peoniflorin and MPP+ than the control groups. There was statistical significant difference between high concentration peoniflorin and control group (P<0.01) by comparing of the amont of TH+ cells.[Conclusion] 1.Basicly, the process of substantia nigra development in the organotypic brain slice culture in vitro is almost similar to that in the nature postnatal development in vivo, except that the development in vitro was more hysteresis. 2.The rat organotypic brain slices culture of substantia nigra was a favourable method to study on Parkinson's disease pathogenesy. 3.High concentration(10.0μmol/L)of Peoniflorin was effective in protecting of dopamine neurons in substantia nigra.