CNTF在青光眼大鼠视网膜中表达变化和对纯化培养大鼠视网膜神经节细胞影响.pdf

THE EXPRESSION VARIATION OF CNTF IN RETINA OF GLAUCOMA RAT AND THE EFFECTS OF CNTF ON PURIFIED RAT RETINAL GANGLION CELLS IN VITRO ABSTRACT Objective (1) To establish a rat glaucoma model. (2)To observe the change of cellular localization of ciliary neurotrophic factor in retina of rat glaucoma model compare with normal rat.(3) To study the effects of ciliary neurotrophic factor on cytoactive and processes regrowth of purified rat retinal ganglion cells in vitro. Methods (1) The rat glaucoma model with chronic increased intraocular pressure which was induced by electrocoagulation of the episcleral and limbal veins. (2) The retinas were harvested st th th in 1 day, 7 day and 28 day after the model was established. Immunohistochemical staining was used to localize the cells those express ciliary neurotrophic factor. (3) SD rat (1～3 days after birth) retinas were dissected and dissociated. Retinal ganglion cells were purified by grit and cultured in 96-wells plates. 5 sorts -1 -1 of culture medium which separately contain 0ng ·ml 、10ng ·ml 、 -1 -1 -1 20ng ·ml 、30ng ·ml 、40ng ·ml ciliary neurotrophic factor were prepared. 24 hours after cells were seeded, 4 sorts of medium which contain ciliary neurotrophic factor were added in, and Retinal ganglion cells cultured without ciliary neurotrophic factor were 3 used as control. The cells were identified by morphologic criteria st rd th th th and immunocytochemical staining. In 1 d, 3 d, 5 d, 7 d, 9 d, th th th 11 d, 13 d, 15 d after ciliary neurotrophic factor was added, MTT colorimetric assay was adopted to determine the