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目的研究碱性成纤维细胞生长因子(bFGF)在持续高眼压下对兔视网膜神经节细胞的保护作用。方法健康新西兰大白兔27只,其中24只(48眼)卡波姆注入前房法制成持续高眼压动物模型,随机分为2组,每只兔一眼为高眼压组(各24眼),另一眼为bFGF实验组(各24眼);另3只(6眼)为正常组。bFGF实验组在建模过程中及成功后1、2、3周玻璃体腔内注射bFGF各1次,正常组在相应的时间仅玻璃体腔内注入与bFGF实验组等体积的磷酸缓冲生理盐水(PBS)。术后每日固定时间测量眼压,将眼压维持在30mmHg以上。分别于建模后1、2、3、4周随机处死6只兔子。完整摘除眼球行常规HE染色观察视网膜神经节细胞形态变化并计数存活细胞数目,免疫组织化学染色检测视网膜神经节细胞Bcl-2凋亡基因的表达。建模后4周行电镜观察视网膜神经节细胞超微结构的变化。结果光镜:建模后两组神经节细胞数目均较正常组减少,自2周后高眼压组与bFGF实验组比较差异有统计学意义(P<0.05)。免疫组织化学:比较视网膜神经节细胞抗凋亡蛋白(RGCsBcl-2)基因蛋白表达的阳性目标面积构成比,第3周时bFGF实验组Bcl-2基因蛋白仍保持较高水平的表达,两组比较差异有统计学意义(P<0.01)。电镜:术后4周bFGF实验组改变明显轻于高眼压组,细胞器较对照组丰富,部分线粒体仍可见完整嵴。结论bFGF作为一种神经营养因子,对高眼压下兔视网膜神经节细胞具有保护作用。
Objective To investigate the protective effect of basic fibroblast growth factor (bFGF) on retinal ganglion cells in rats under continuous ocular hypertension. Methods Twenty-seven New Zealand white rabbits were randomly divided into 2 groups, each with 24 eyes (48 eyes) with intraocular pressure (24 eyes each) , Another eye bFGF experimental group (24 eyes each); the other three (6 eyes) were normal group. bFGF experimental group in the modeling process and 1, 2, 3 weeks after successful injection of bFGF vitreous cavity one time, the normal group at the appropriate time only intravitreal injection with bFGF experimental group equal volume of phosphate buffered saline (PBS ). Intraocular pressure was measured daily after a fixed time, the intraocular pressure was maintained at 30mmHg or more. Six rabbits were randomly sacrificed at 1, 2, 3 and 4 weeks after modeling. The retinal ganglion cells were removed by HE staining. The number of surviving cells was counted. The expression of Bcl-2 gene was detected by immunohistochemical staining. Changes of retinal ganglion cell ultrastructure were observed under electron microscope 4 weeks after modeling. Results Light microscope: After modeling, the number of ganglion cells in both groups decreased compared with that in normal group. There was significant difference between experimental group and bFGF experimental group after 2 weeks (P <0.05). Immunohistochemistry: Compared with the positive target area ratio of retinal ganglion cell anti-apoptotic protein (RGCsBcl-2) gene expression, the expression of Bcl-2 protein in bFGF experimental group remained high at the third week. The difference was statistically significant (P <0.01). Electron microscopy: At 4 weeks after operation, the changes of bFGF experimental group were obviously lighter than those of high intraocular pressure group. The organelles were more abundant than the control group, and some mitochondria still showed complete ridges. Conclusion bFGF, a neurotrophic factor, has a protective effect on rabbit retinal ganglion cells under ocular hypertension.