基因转染的虹膜色素上皮细胞治疗RCS大鼠视网膜色素变性的实验研究
发布时间：2018-08-31 20:06
【摘要】：遗传性视网膜变性是一组以进行性光感受器细胞及色素上皮细胞功能丧失为共同表现的疾病。目前为止,这些疾病尚无有效治疗方法[1, 2]。全世界有数百万人因此永久致盲,危害性极大。临床上迫切需要有效措施以改善和维护RPE功能,防止感光细胞变性及凋亡,这是多年来全世界范围内一大热点和难点。目前发现脑源性神经营养因子(brain derived neurotrophic factor, BDNF)能促进培养的人和牛的视网膜色素上皮(retinal pigment epithelium, RPE)细胞分化,通过自分泌或通过胶质细胞调节视网膜变性中感光细胞的生存,使视网膜细胞存活[3, 4, 5]。 本研究拟在视网膜变性的RCS(Royal College of Surgeons)鼠早期进行干预,将BDNF基因转染的虹膜色素上皮细胞(iris pigment epitheium,IPE)(AAV-BDNF-IPE)移植入早期RCS大鼠视网膜下腔后,观察不同时期视网膜电图(electroretinogram, ERG)、视网膜组织BDNF表达及视网膜形态变化,分析移植后RCS鼠视网膜改善情况,为临床对由于RPE功能障碍以及感光细胞变性引起的疾病患者提供更多的思路。 方法本研究通过以下四部分进行: 第一部分:出生后18-22天(3周龄)RCS大鼠48只,雌雄不拘。通过改进的巩膜外路移植法将AAV-BDNF-IPE细胞移植到RCS大鼠双眼视网膜下腔,术后观察眼底情况,并行HE染色,观察移植细胞的存活情况。 第二部分:SD正常对照大鼠,按照出生后3周龄、6、8、10、12、14周龄分组,每周龄组3只,共18只,RCS大鼠及AAV-BDNF-IPE细胞移植RCS大鼠(已在第一部分实验中完成AAV-BDNF-IPE细胞移植)各18只,按出生后周龄分组同SD大鼠,分别行双眼暗适应ERG记录,电生理检查参照国际临床视觉电生理标准。每次每眼至少检查3次,叠加后取波形稳定、干扰小的ERG图形,计算ERG-a、b波振幅值并进行统计学处理。 第三部分:对照组RCS大鼠按照出生后3周龄、6、8、10、12、14周龄分组,每周龄组4只,共24只;AAV-BDNF-IPE细胞移植RCS大鼠(已在第一部分实验中完成AAV-BDNF-IPE细胞移植)24只,按出生后周龄分组同对照组RCS大鼠。分别于相应时间点即手术当天、术后3、5、7、9、11周取双眼球,用酶联免疫吸附法(Elisa)检测视网膜组织中BDNF的表达水平,比较分析这些数据并进行统计学处理。 第四部分:实验动物及分组同第二部分,行电生理检测后处死大鼠,取出双眼球,取视网膜组织进行HE染色、TUNEL染色和免疫组化染色检测。 结果 1.共计43只RCS大鼠实施了双眼视网膜下腔移植,发生大量玻璃体积血、广泛视网膜脱离、眼内炎及误注入玻璃体腔18眼。AAV-BDNF-IPE细胞成功移植入视网膜下腔68眼,手术成功率79%。 RCS大鼠视网膜及睫状体部位HE染色发现,睫状体及视网膜均缺乏色素细胞层;AAV-BDNF-IPE细胞移植到RCS大鼠视网膜下腔5周后,HE染色可见大鼠视网膜下腔有散在存活的褐色上皮细胞,推测为移植后存活的AAV-BDNF-IPE细胞;视网膜或脉络膜内未见明显炎性细胞浸润反应及结构破坏(Fig.6)。 2.三组大鼠成功完成暗适应FERG记录,从出生后第3周到第14周,共记录正常SD大鼠15只30次,未干预对照RCS大鼠15只30次,细胞移植组RCS大鼠17只34次。 三组大鼠生后8周内FERG的暗适应最大反应a、b波振幅无明显差别,出生10周以后RCS大鼠FERG的暗适应最大反应a、b波振幅均明显降低,与同周龄正常SD大鼠相比显著下降(P㩳0.05),出生12周以后RCS大鼠ERG波形呈低平型,未见明显波峰波谷,a、b波模糊。AAV-BDNF-IPE移植手术组RCS大鼠出生后10周龄时b波仍保持较高水平,尽管细胞移植组RCS大鼠b波呈现缓慢下降趋势,但振幅仍显著高于同周龄未干预对照组RCS大鼠(12w和14w组P0.05)。 3.对照组RCS大鼠出生后3周龄时视网膜组织中BDNF仍保持较高水平,其后迅速降低,其中3周龄组与其它周龄组比较,P0.01;手术组RCS大鼠术时、术后3、5、7、9、11周各组间两两比较,BDNF表达无显著差异(P㧐0.05);出生后6周龄直到14周龄的不同时期,AAV-BDNF-IPE移植手术组RCS大鼠视网膜BDNF表达水平均明显高于对照组(其中6周龄组P0.05,其它各周龄组P0.01)。 4. HE染色结果示:AAV-BDNF-IPE移植手术组和对照组相比视杆层靠近外核层的一侧厚度略有改善;TUNEL结果:实验组凋亡指数减少,凋亡峰值后延,凋亡高峰发生于移植术后9wk;免疫组化染色结果示:实验组大鼠原外层视网膜部位GFAP呈弱阳性表达,Muller细胞着色较浅;SYN除内丛状层着色外,外丛状层呈弱阳性反应。 小结 1、本实验采用自制的注射器,经单通道巩膜外路法成功将AAV-BDNF-IPE移植至幼年RCS大鼠视网膜下腔,有效缩小了巩膜切口,减少了视网膜解剖结构的破坏,手术成功率达到79%。 RCS大鼠视网膜及睫状体部位HE染色发现,睫状体及视网膜均缺乏色素细胞层;AAV-BDNF-IPE细胞移植到RCS大鼠视网膜下腔5周后,HE染色可见大鼠视网膜下腔有散在存活的褐色上皮细胞,推测为移植后存活的AAV-BDNF-IPE细胞;视网膜或脉络膜内未见明显炎性细胞浸润反应及结构破坏。 虽然不能判定移植细胞与其宿主视网膜组织间建立了正确的解剖联系,但我们的实验结果证实,通过改进的巩膜外路移植法将AAV-BDNF-IPE细胞移植至视网膜下腔,移植细胞不但能够较长期存活,而且不发生免疫排斥反应,是安全可靠的。 2、本实验发现SD正常对照大鼠、RCS大鼠及AAV-BDNF-IPE细胞移植RCS大鼠三组大鼠生后8周内FERG的暗适应最大反应a、b波振幅无明显差别,出生10周以后RCS大鼠FERG的暗适应最大反应a、b波振幅均明显降低,与同周龄正常SD大鼠相比显著下降(P㩳0.05),出生12周以后RCS大鼠ERG波形呈低平型,未见明显波峰波谷,a、b波模糊,这些结果证实RCS大鼠视网膜色素变性进展过程中,视网膜功能逐步丧失。RCS大鼠视网膜电图的改变滞后于视网膜形态改变。 将AAV-BDNF-IPE细胞移植入RCS大鼠视网膜下腔后,可以明显改善FERG b波振幅,尽管随着术后时间延长,细胞移植组RCS大鼠b波呈现缓慢下降趋势,但出生后10周龄后b波仍保持相对较高水平,且振幅显著高于同周龄未干预对照组RCS大鼠(12w和14w组P0.05),提示AAV-BDNF-IPE细胞移植可以在一定程度上延缓RCS大鼠视网膜色素变性的进展,改善部分视网膜功能。 3.对照组RCS大鼠出生后3周龄时视网膜组织中BDNF仍保持较高水平,其后迅速降低;出生后6周龄直到14周龄的不同时期,AAV-BDNF-IPE移植手术组RCS大鼠视网膜BDNF表达水平均明显高于对照组。这些结果表明,BDNF基因转染的虹膜色素上皮细胞在RCS大鼠视网膜下腔移植后,视网膜组织中BDNF可以持续稳定高水平表达,这是AAV-BDNF-IPE移植改善宿主视网膜功能的一个重要原因。 4. AAV-BDNF-IPE细胞移植效果比未干预组略好:视杆层靠近外核层的一侧厚度也略有改善,这与以往的研究一致;TUNEL检测证实视细胞的丧失是一种凋亡现象,也提示AAV-BDNF-IPE的治疗可能是抑制细胞凋亡而发挥作用的;免疫组化结果提示可能减轻了代偿反应性胶质纤维增生,也可能表现了以突触恢复为特征的超微结构的变化。 综上所述,该实验延缓了视网膜变性的进一步发展,表明这一方法可能拯救变性的光感受器,使之重新恢复功能。
[Abstract]:Hereditary retinal degeneration is a group of diseases characterized by progressive photoreceptor cell and pigment epithelial cell dysfunction. Up to now, there is no effective treatment for these diseases [1,2]. Preventing degeneration and apoptosis of photoreceptor cells has been a hot and difficult issue worldwide for many years. Brain derived neurotrophic factor (BDNF) has been found to promote the differentiation of cultured human and bovine retinal pigment epithelium (RPE) cells through autocrine or glial fineness. The cells regulate the survival of photoreceptors in retinal degeneration, so that retinal cells survive [3, 4, 5].
This study was designed to observe the expression of BDNF and electroretinogram (ERG) in the retina of early RCS rats after transplantation of iris pigment epithelium (IPE) transfected with BDNF gene into the subretinal space of early RCS rats. The morphological changes of the retina and the improvement of the retina in RCS mice after transplantation were analyzed to provide more ideas for the clinical treatment of patients with RPE dysfunction and photoreceptor degeneration.
Methods the study was carried out in the following four parts:
The first part: 48 RCS rats aged from 18 to 22 days after birth, male and female. AAV-BDNF-IPE cells were transplanted into the subretinal space of both eyes of RCS rats by improved scleral transplantation. The fundus of the eyes was observed after operation, and the survival of the transplanted cells was observed by HE staining.
The second part: SD normal control rats were divided into 3 weeks old, 6,8,10,12,14 weeks old, 3 rats in each age group, 18 rats in each age group. RCS rats and 18 RCS rats transplanted with AAV-BDNF-IPE cells (which had been transplanted with AAV-BDNF-IPE cells in the first experiment) were divided into two groups according to the age of birth and SD rats were recorded by binocular dark adaptation ERG. Physical examination refers to the international clinical visual electrophysiology standard. Each eye is examined at least three times. After stacking, ERG patterns with stable waveform and little interference are obtained. ERG-a and b wave amplitudes are calculated and statistically processed.
The third part: The control group RCS rats were divided into 3 weeks old, 6,8,10,12,14 weeks old, 4 rats in each week group, 24 rats in total; 24 RCS rats transplanted with AAV-BDNF-IPE cells (which had been transplanted into AAV-BDNF-IPE cells in the first part of the experiment), and the control group RCS rats were divided into two groups according to the age after birth. The expression of BDNF in retinal tissue was detected by enzyme linked immunosorbent assay (Elisa) at 7,9 and 11 weeks after operation.
The fourth part: The experimental animals and groups were the same as the second part. The rats were sacrificed after electrophysiological examination. The eyeballs were taken out and the retinal tissues were stained with HE, TUNEL and immunohistochemical staining.
Result
1. A total of 43 RCS rats underwent bilateral subretinal cavity transplantation, which resulted in massive vitreous hemorrhage, extensive retinal detachment, endophthalmitis and misinjection into the vitreous cavity in 18 eyes. AAV-BDNF-IPE cells were successfully transplanted into the subretinal cavity in 68 eyes, with a success rate of 79%.
HE staining in the retina and ciliary body of RCS rats showed that there was no pigment cell layer in both ciliary body and retina. After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats for 5 weeks, HE staining showed that there were scattered surviving Brown epithelial cells in the subretinal space of the rats, presumed to be surviving AV-BDNF-IPE cells in the retina or choroid. No obvious inflammatory cell infiltration and structural damage were observed (Fig.6).
2. Three groups of rats successfully completed dark adaptation FERG recording. From the 3rd week to the 14th week after birth, 15 normal SD rats were recorded 30 times, 15 non-intervention RCS rats 30 times, and 17 RCS rats 34 times in cell transplantation group.
There was no significant difference in the amplitudes of a and B waves of the maximum dark adaptation response of FERG among the three groups within 8 weeks after birth. The amplitudes of a and B waves of the maximum dark adaptation response of FERG in RCS rats decreased significantly after 10 weeks of birth compared with normal SD rats of the same age (P?0.05). After 12 weeks of birth, the ERG waveforms of RCS rats showed a low flat pattern, and no obvious peaks, troughs, A and B modes. The B wave of RCS rats in AAV-BDNF-IPE transplantation group remained high at 10 weeks after birth. Although the B wave of RCS rats in cell transplantation group showed a slowly decreasing trend, the amplitude of B wave was still significantly higher than that of RCS rats in non-intervention control group (P 0.05 in 12W and 14W groups).
3. The levels of BDNF in the retina of RCS rats in the control group remained high at 3 weeks after birth, and then decreased rapidly. Compared with other age groups, the levels of BDNF in the retina of RCS rats in the 3-week-old group were P 0.01; there was no significant difference in the expression of BDNF between the two groups at 3, 5, 7, 9 and 11 weeks after operation (P? The expression of BDNF in the retina of RCS rats after BDNF-IPE transplantation was significantly higher than that in the control group (P 0.05 in the 6-week-old group and P 0.01 in the other groups).
4. HE staining showed that the thickness of the optic rod layer near the outer nuclear layer was slightly improved in AAV-BDNF-IPE transplantation group compared with the control group; TUNEL results: the apoptosis index of the experimental group was reduced, the apoptosis peak was delayed, and the apoptosis peak occurred 9 wk after transplantation; immunohistochemical staining showed that GFAP was weakly positive in the original outer retina of the experimental group rats. The expression of Muller cells was relatively pale, while the outer plexiform layer of SYN was weakly positive except for the inner plexiform layer.
Summary
1. AAV-BDNF-IPE was successfully transplanted into the subretinal space of young RCS rats by single-channel scleral approach with a self-made injector. The scleral incision was effectively reduced and the damage of retinal anatomy was reduced. The success rate of the operation was 79%.
HE staining in the retina and ciliary body of RCS rats showed that there was no pigment cell layer in both ciliary body and retina. After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats for 5 weeks, HE staining showed that there were scattered surviving Brown epithelial cells in the subretinal space of the rats, presumed to be surviving AV-BDNF-IPE cells in the retina or choroid. No obvious inflammatory cell infiltration and structural damage were observed.
Although it is not possible to determine the correct anatomical relationship between the transplanted cells and their host retinal tissues, our experimental results confirm that the transplantation of AAV-BDNF-IPE cells into the subretinal space by improved scleral transplantation is safe and reliable, not only for long-term survival, but also for the absence of immune rejection.
2. In this study, we found that there was no significant difference in the amplitude of a and B waves of the maximum dark adaptation response of FERG between the three groups of SD normal control rats, RCS rats and AAV-BDNF-IPE cells transplanted RCS rats within 8 weeks after birth. After 10 weeks of birth, the amplitude of a and B waves of the maximum dark adaptation response of FERG in RCS rats decreased significantly, compared with normal SD rats of the same age (P?0.05). After 12 weeks of birth, the ERG waveforms of RCS rats were low and flat, and there were no obvious peaks and troughs, and a and B waves were blurred. These results confirmed that the retinal function was gradually lost during the development of RPD in RCS rats.
After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats, the amplitude of FERG-b wave could be significantly improved. Although the amplitude of B wave in RCS rats decreased slowly with the prolonged time after transplantation, the amplitude of B wave remained relatively high after 10 weeks of birth, and was significantly higher than that in RCS rats without intervention at the same age (12 weeks and 14 weeks). Group P 0.05), suggesting that AAV-BDNF-IPE cell transplantation can delay the progress of retinitis pigmentosa in RCS rats to some extent and improve some retinal function.
3. BDNF levels in the retina of RCS rats in the control group remained high at 3 weeks after birth, and then decreased rapidly; BDNF expression levels in the retina of RCS rats in the AAV-BDNF-IPE transplantation group were significantly higher than those in the control group from 6 weeks to 14 weeks after birth. After transplantation of RCS rats into the subretinal space, the expression of BDNF in the retina can be maintained at a high level, which is an important reason why AAV-BDNF-IPE transplantation can improve the function of the host retina.
4. The transplantation effect of AAV-BDNF-IPE cells was slightly better than that of non-intervention group: the thickness of rod layer near the outer nuclear layer was also slightly improved, which was consistent with previous studies; TUNEL detection confirmed that loss of optic cells was an apoptotic phenomenon, and suggested that AAV-BDNF-IPE treatment might play a role in inhibiting apoptosis; immunohistochemical results suggested that It may alleviate the compensatory reactive glial fibrillary hyperplasia and may also show the ultrastructural changes characterized by synaptic recovery.
In summary, this experiment delayed the further development of retinal degeneration, suggesting that this method may save degenerated photoreceptors and restore their function.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1
本文编号：2216023
[Abstract]:Hereditary retinal degeneration is a group of diseases characterized by progressive photoreceptor cell and pigment epithelial cell dysfunction. Up to now, there is no effective treatment for these diseases [1,2]. Preventing degeneration and apoptosis of photoreceptor cells has been a hot and difficult issue worldwide for many years. Brain derived neurotrophic factor (BDNF) has been found to promote the differentiation of cultured human and bovine retinal pigment epithelium (RPE) cells through autocrine or glial fineness. The cells regulate the survival of photoreceptors in retinal degeneration, so that retinal cells survive [3, 4, 5].
This study was designed to observe the expression of BDNF and electroretinogram (ERG) in the retina of early RCS rats after transplantation of iris pigment epithelium (IPE) transfected with BDNF gene into the subretinal space of early RCS rats. The morphological changes of the retina and the improvement of the retina in RCS mice after transplantation were analyzed to provide more ideas for the clinical treatment of patients with RPE dysfunction and photoreceptor degeneration.
Methods the study was carried out in the following four parts:
The first part: 48 RCS rats aged from 18 to 22 days after birth, male and female. AAV-BDNF-IPE cells were transplanted into the subretinal space of both eyes of RCS rats by improved scleral transplantation. The fundus of the eyes was observed after operation, and the survival of the transplanted cells was observed by HE staining.
The second part: SD normal control rats were divided into 3 weeks old, 6,8,10,12,14 weeks old, 3 rats in each age group, 18 rats in each age group. RCS rats and 18 RCS rats transplanted with AAV-BDNF-IPE cells (which had been transplanted with AAV-BDNF-IPE cells in the first experiment) were divided into two groups according to the age of birth and SD rats were recorded by binocular dark adaptation ERG. Physical examination refers to the international clinical visual electrophysiology standard. Each eye is examined at least three times. After stacking, ERG patterns with stable waveform and little interference are obtained. ERG-a and b wave amplitudes are calculated and statistically processed.
The third part: The control group RCS rats were divided into 3 weeks old, 6,8,10,12,14 weeks old, 4 rats in each week group, 24 rats in total; 24 RCS rats transplanted with AAV-BDNF-IPE cells (which had been transplanted into AAV-BDNF-IPE cells in the first part of the experiment), and the control group RCS rats were divided into two groups according to the age after birth. The expression of BDNF in retinal tissue was detected by enzyme linked immunosorbent assay (Elisa) at 7,9 and 11 weeks after operation.
The fourth part: The experimental animals and groups were the same as the second part. The rats were sacrificed after electrophysiological examination. The eyeballs were taken out and the retinal tissues were stained with HE, TUNEL and immunohistochemical staining.
Result
1. A total of 43 RCS rats underwent bilateral subretinal cavity transplantation, which resulted in massive vitreous hemorrhage, extensive retinal detachment, endophthalmitis and misinjection into the vitreous cavity in 18 eyes. AAV-BDNF-IPE cells were successfully transplanted into the subretinal cavity in 68 eyes, with a success rate of 79%.
HE staining in the retina and ciliary body of RCS rats showed that there was no pigment cell layer in both ciliary body and retina. After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats for 5 weeks, HE staining showed that there were scattered surviving Brown epithelial cells in the subretinal space of the rats, presumed to be surviving AV-BDNF-IPE cells in the retina or choroid. No obvious inflammatory cell infiltration and structural damage were observed (Fig.6).
2. Three groups of rats successfully completed dark adaptation FERG recording. From the 3rd week to the 14th week after birth, 15 normal SD rats were recorded 30 times, 15 non-intervention RCS rats 30 times, and 17 RCS rats 34 times in cell transplantation group.
There was no significant difference in the amplitudes of a and B waves of the maximum dark adaptation response of FERG among the three groups within 8 weeks after birth. The amplitudes of a and B waves of the maximum dark adaptation response of FERG in RCS rats decreased significantly after 10 weeks of birth compared with normal SD rats of the same age (P?0.05). After 12 weeks of birth, the ERG waveforms of RCS rats showed a low flat pattern, and no obvious peaks, troughs, A and B modes. The B wave of RCS rats in AAV-BDNF-IPE transplantation group remained high at 10 weeks after birth. Although the B wave of RCS rats in cell transplantation group showed a slowly decreasing trend, the amplitude of B wave was still significantly higher than that of RCS rats in non-intervention control group (P 0.05 in 12W and 14W groups).
3. The levels of BDNF in the retina of RCS rats in the control group remained high at 3 weeks after birth, and then decreased rapidly. Compared with other age groups, the levels of BDNF in the retina of RCS rats in the 3-week-old group were P 0.01; there was no significant difference in the expression of BDNF between the two groups at 3, 5, 7, 9 and 11 weeks after operation (P? The expression of BDNF in the retina of RCS rats after BDNF-IPE transplantation was significantly higher than that in the control group (P 0.05 in the 6-week-old group and P 0.01 in the other groups).
4. HE staining showed that the thickness of the optic rod layer near the outer nuclear layer was slightly improved in AAV-BDNF-IPE transplantation group compared with the control group; TUNEL results: the apoptosis index of the experimental group was reduced, the apoptosis peak was delayed, and the apoptosis peak occurred 9 wk after transplantation; immunohistochemical staining showed that GFAP was weakly positive in the original outer retina of the experimental group rats. The expression of Muller cells was relatively pale, while the outer plexiform layer of SYN was weakly positive except for the inner plexiform layer.
Summary
1. AAV-BDNF-IPE was successfully transplanted into the subretinal space of young RCS rats by single-channel scleral approach with a self-made injector. The scleral incision was effectively reduced and the damage of retinal anatomy was reduced. The success rate of the operation was 79%.
HE staining in the retina and ciliary body of RCS rats showed that there was no pigment cell layer in both ciliary body and retina. After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats for 5 weeks, HE staining showed that there were scattered surviving Brown epithelial cells in the subretinal space of the rats, presumed to be surviving AV-BDNF-IPE cells in the retina or choroid. No obvious inflammatory cell infiltration and structural damage were observed.
Although it is not possible to determine the correct anatomical relationship between the transplanted cells and their host retinal tissues, our experimental results confirm that the transplantation of AAV-BDNF-IPE cells into the subretinal space by improved scleral transplantation is safe and reliable, not only for long-term survival, but also for the absence of immune rejection.
2. In this study, we found that there was no significant difference in the amplitude of a and B waves of the maximum dark adaptation response of FERG between the three groups of SD normal control rats, RCS rats and AAV-BDNF-IPE cells transplanted RCS rats within 8 weeks after birth. After 10 weeks of birth, the amplitude of a and B waves of the maximum dark adaptation response of FERG in RCS rats decreased significantly, compared with normal SD rats of the same age (P?0.05). After 12 weeks of birth, the ERG waveforms of RCS rats were low and flat, and there were no obvious peaks and troughs, and a and B waves were blurred. These results confirmed that the retinal function was gradually lost during the development of RPD in RCS rats.
After transplantation of AAV-BDNF-IPE cells into the subretinal space of RCS rats, the amplitude of FERG-b wave could be significantly improved. Although the amplitude of B wave in RCS rats decreased slowly with the prolonged time after transplantation, the amplitude of B wave remained relatively high after 10 weeks of birth, and was significantly higher than that in RCS rats without intervention at the same age (12 weeks and 14 weeks). Group P 0.05), suggesting that AAV-BDNF-IPE cell transplantation can delay the progress of retinitis pigmentosa in RCS rats to some extent and improve some retinal function.
3. BDNF levels in the retina of RCS rats in the control group remained high at 3 weeks after birth, and then decreased rapidly; BDNF expression levels in the retina of RCS rats in the AAV-BDNF-IPE transplantation group were significantly higher than those in the control group from 6 weeks to 14 weeks after birth. After transplantation of RCS rats into the subretinal space, the expression of BDNF in the retina can be maintained at a high level, which is an important reason why AAV-BDNF-IPE transplantation can improve the function of the host retina.
4. The transplantation effect of AAV-BDNF-IPE cells was slightly better than that of non-intervention group: the thickness of rod layer near the outer nuclear layer was also slightly improved, which was consistent with previous studies; TUNEL detection confirmed that loss of optic cells was an apoptotic phenomenon, and suggested that AAV-BDNF-IPE treatment might play a role in inhibiting apoptosis; immunohistochemical results suggested that It may alleviate the compensatory reactive glial fibrillary hyperplasia and may also show the ultrastructural changes characterized by synaptic recovery.
In summary, this experiment delayed the further development of retinal degeneration, suggesting that this method may save degenerated photoreceptors and restore their function.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1
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