超声微泡造影剂介导脑源性神经营养因子联合转染视网膜和视皮质对视神经损伤后神经节细胞的保护作用

发布时间：2018-05-04 06:30

本文选题：脑源性神经营养因子 + 视网膜神经节细胞　；参考：《重庆医科大学》2011年硕士论文

【摘要】：目的观察超声微泡造影剂介导脑源性神经营养因子(BDNF)联合转染大鼠视网膜和视皮质区细胞对视神经损伤后视网膜神经节细胞(RGC)的保护作用。方法雄性Sprague-Dawley(SD)大鼠88只随机分为正常组(A组)、假手术组(B组)、空白对照组(C组)、单纯眼转染组(D组)、单纯脑转染组(E组)、联合转染组(F组)。A组8只大鼠,B～F组每组16只大鼠。建立钳夹视神经损伤模型,将B～F组随机分为视神经损伤1、2周亚组,各亚组8只大鼠。B、C组分别玻璃体腔和视皮质区注射磷酸盐缓冲液(PBS),D、E组分别玻璃体腔和视皮质区注射BDNF质粒(pBDNF)微泡造影剂悬液,F组玻璃体腔和视皮质区同时注射pBDNF微泡造影剂悬液。D～F组注射pBDNF微泡微泡造影剂悬液后,立即用超声辐照相应转染部位。视神经损伤1、2周,各组行逆行荧光金标记RGC计数;半胱氨酸蛋白酶-3(caspase-3)蛋白免疫组织化学染色,观察其阳性表达情况;视网膜电流图(PERG)检测,记录N95振幅。结果荧光金标记检测显示F组视神经损伤1、2周亚组RGC数分别为409.1±42.10个、390.8±35.34个,占A组的86.0%、82.1%,均明显高于C、D、E组,差异具有统计学意义(P㩳0.01)。F组各时间点Caspase-3阳性细胞数均少于C、D、E组,且染色较浅。PERG检测中视神经损伤1周、2周时F组N95振幅分别为7.57±0.66μV、7.33±0.58μV,高于C、D、E组(P㩳0.01),且与A组无明显差异(P0.05)。结论超声微泡造影剂介导BDNF基因联合转染视网膜和视皮质区细胞能抑制视神经损伤后RGC凋亡,提高RGC存活数,还可能具有一定的电生理功能保护作用。
[Abstract]:Purpose To observe the protective effect of combined transfection of brain derived neurotrophic factor (BDNF) with ultrasound microbubble contrast medium on retinal ganglion cells (RGCs) after optic nerve injury in rats. Method 88 male Sprague-Dawley SD rats were randomly divided into normal group (n = 8), sham operation group (n = 8), control group (n = 16), control group (n = 8), group D (n = 8), group E (n = 8), group A (n = 8), group B (n = 8). The model of optic nerve injury by clamp was established, and the group B F was randomly divided into the group of 1 ~ 2 weeks after optic nerve injury. Eight rats in each subgroup were injected with BDNF plasmid pBDNF in vitreous body cavity and visual cortex, respectively. PBDNF was injected into vitreous body cavity and visual cortex respectively. PBDNF was injected into visual cortex and vitreous cavity of F group. After injection of pBDNF microbubble contrast agent suspension, the microbubble contrast agent suspension. The corresponding transfection site was immediately irradiated by ultrasound. After 1 and 2 weeks of optic nerve injury, the number of RGC labeled with retrograde fluorescence gold, the immunohistochemical staining of cysteine protease -3caspase-3) and the positive expression of cysteine protease -3caspase-3 were observed in each group, and the amplitude of N95 was recorded by electroretinogram (ERG). Result The number of RGC in group F was 409.1 卤42.10 (390.8 卤35.34) at 1 and 2 weeks after optic nerve injury, which accounted for 82.1% of group A, which was significantly higher than that in group E (P < 0.01). The number of Caspase-3 positive cells in group F was lower than that in group E at each time point. The N95 amplitudes of group F were 7.57 卤0.66 渭 V, 7.33 卤0.58 渭 V, respectively, which were higher than those of group A (P 0.01), and there was no significant difference between group A and group A. Conclusion The combined transfection of BDNF gene into retinal and visual cortex cells mediated by ultrasound microbubble contrast agent can inhibit the apoptosis of RGC after optic nerve injury and increase the survival of RGC. It may also have a protective effect on electrophysiological function.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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