脂肪干细胞玻璃体腔移植对大鼠青光眼模型视网膜神经节细胞的保护作用

发布时间：2018-06-24 04:21

本文选题：脂肪干细胞 + 青光眼　；参考：《青岛大学》2017年硕士论文

【摘要】：目的:研究脂肪干细胞玻璃体腔注射对大鼠青光眼模型的视网膜神经节细胞的保护作用,并初步探讨可能的机制。方法:培养脂肪干细胞,培养至P4接近单层融合时,收集并调整细胞密度备用。将大鼠分为四组:青光眼干细胞组、青光眼对照组、正常眼压干细胞组和正常眼压对照组,采用烙闭表层巩膜静脉的方法制备大鼠青光眼模型,均以右眼为操作眼,青光眼干细胞组、青光眼对照组行造模手术,正常眼压干细胞组、正常眼压对照组行假手术操作。手术操作3周时,三组均行玻璃体腔注射操作,青光眼干细胞组注射物为5×105 ADSCs悬浮液6?L,青光眼对照组注射物为DMEM液6?L,正常眼压对照组注射物为5×105 ADSCs悬浮液6?L,正常眼压对照组注射物DMEM液6?L。分别测量并记录术前、术后1周、2周、3周、4周、5周、6周眼压。实验完成后将动物处死,随机摘取眼球,分别进行冰冻切片HE染色、免疫荧光染色处理,并进行q RT-PCR检测BDNF m RNA表达量。结果:造模后1周,模型眼相对于假手术眼,IOP平均升高10.9±2.1mm Hg,并能在预定实验时间6周内保持这一水平。而在对各组进行移植操作后,青光眼干细胞组的IOP降低水平在移植操作后1、2、3周时均显著高于青光眼对照组(p0.001)。HE染色青光眼干细胞组、青光眼对照组与正常眼压干细胞组、正常眼压对照组相较均出现后极部视网膜各层变薄,RGC层细胞排列紊乱,RGC数量减少,但青光眼对照组较青光眼干细胞组病理改变严重。免疫荧光染色两组青光眼组与两组正常眼压组相较,荧光染色淡,神经节细胞数明显减少,青光眼对照组较青光眼干细胞组减少更明显。除正常眼压两组间外,其他各组间在400倍荧光显微镜下RGC计数差异均具有统计学意义,计数:青光眼对照组青光眼干细胞组正常眼压干细胞组/正常眼压对照组。BDNF m RNA各组之间差异具有统计学意义(*P0.05;**P0.01;***P0.001),两两比较差异具有统计学意义,且BDNF m RNA表达量:青光眼干细胞组正常眼压干细胞组青光眼对照组正常眼压对照组。结论:脂肪干细胞玻璃体腔移植对大鼠青光眼模型视网膜神经节细胞具有保护作用,可能通过降低眼压和增多BDNF等途径发挥保护作用。
[Abstract]:Aim: to study the protective effect of adipose stem cells intravitreal injection on retinal ganglion cells (RGCs) in rat glaucoma model and to explore the possible mechanism. Methods: adipose stem cells were cultured until P 4 was close to monolayer fusion. Cell density was collected and adjusted. Rats were divided into four groups: glaucoma stem cell group, glaucoma control group, normal intraocular pressure stem cell group and normal intraocular pressure control group. Glaucoma stem cell group, glaucoma control group, normal IOP stem cell group, normal IOP control group were performed sham operation. After 3 weeks of operation, all the three groups were treated with intravitreal injection. In the glaucoma stem cell group, 5 脳 10 5 ADSCs suspension was 6L, the glaucoma control group was DMEM 6L, the normal intraocular pressure control group was 5 脳 105 ADSCs suspension 6L, and the normal intraocular pressure control group was DMEM 6L. Intraocular pressure was measured and recorded 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks and 6 weeks after operation. After the experiment, the animals were killed, eyeballs were randomly removed, frozen sections were stained with HE and immunofluorescence staining, and the expression of BDNF mRNA was detected by qRT-PCR. Results: the IOP of the model eyes increased by 10.9 卤2.1mm Hg 1 week after the model compared with the sham operation eyes, and could be maintained within 6 weeks of the scheduled experimental time. After transplantation, IOP levels in glaucoma stem cell group were significantly higher than those in glaucoma control group (p0.001), glaucoma control group and normal IOP stem cell group. Compared with the normal intraocular pressure control group, the number of RGC cells in the RGC layer of the posterior polar retinal thinning was decreased, but the pathological changes in the glaucoma control group were more serious than those in the glaucoma stem cell group. Compared with normal intraocular pressure group, immunofluorescence staining showed that the number of ganglion cells in glaucoma group was less than that in normal intraocular pressure group, and the number of ganglion cells in glaucoma control group was significantly lower than that in glaucoma stem cell group. With the exception of normal intraocular pressure (IOP), there were significant differences in RGC counts between the other groups under a 400-fold fluorescence microscope. Counting: the difference of BDNF mRNA between normal IOP stem cell group and normal IOP control group was statistically significant (P0.05, P0.01, P0.001), and the difference between two groups was statistically significant. The expression of BDNF mRNA: normal IOP stem cell group, glaucoma control group, normal IOP control group. Conclusion: vitreous transplantation of adipose stem cells has protective effect on retinal ganglion cells in rat model of glaucoma, which may play a protective role by reducing intraocular pressure and increasing BDNF.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R775;R-332

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