PDF(Pubmed)
Abstract：
UNASSIGNED: ELAVL1/HuR is a keystone regulator of gene expression at the posttranscriptional level, including stress response and homeostasis maintenance. The aim of this study was to evaluate the impact of hur silencing on the age-related degeneration of retinal ganglion cells (RGC), which potentially describes the efficiency of endogenous neuroprotection mechanisms, as well as to assess the exogenous neuroprotection capacity of hur-silenced RGC in the rat glaucoma model.
UNASSIGNED: The study consisted of in vitro and in vivo approaches. In vitro, we used rat B-35 cells to investigate, whether AAV-shRNA-HuR delivery affects survival and oxidative stress markers under temperature and excitotoxic insults. In vivo approach consisted of two different settings. In first one, 35 eight-week-old rats received intravitreal injection of AAV-shRNA-HuR or AAV-shRNA scramble control. Animals underwent electroretinography tests and were sacrificed 2, 4 or 6 months after injection. Retinas and optic nerves were collected and processed for immunostainings, electron microscopy and stereology. For the second approach, animals received similar gene constructs. To induce chronic glaucoma, 8 weeks after AAV injection, unilateral episcleral vein cauterization was performed. Animals from each group received intravitreal injection of metallothionein II. Animals underwent electroretinography tests and were sacrificed 8 weeks later. Retinas and optic nerves were collected and processed for immunostainings, electron microscopy and stereology.
UNASSIGNED: Silencing of hur induced apoptosis and increased oxidative stress markers in B-35 cells. Additionally, shRNA treatment impaired the cellular stress response to temperature and excitotoxic insults. In vivo, RGC count was decreased by 39% in shRNA-HuR group 6 months after injection, when compared to shRNA scramble control group. In neuroprotection study, the average loss of RGCs was 35% in animals with glaucoma treated with metallothionein and shRNA-HuR and 11.4% in animals with glaucoma treated with metallothionein and the scramble control shRNA. An alteration in HuR cellular content resulted in diminished photopic negative responses in the electroretinogram.
UNASSIGNED: Based on our findings, we conclude that HuR is essential for the survival and efficient neuroprotection of RGC and that the induced alteration in HuR content accelerates both the age-related and glaucoma-induced decline in RGC number and function, further confirming HuR\'s key role in maintaining cell homeostasis and its possible involvement in the pathogenesis of glaucoma.
UNASSIGNED: The study consisted of in vitro and in vivo approaches. In vitro, we used rat B-35 cells to investigate, whether AAV-shRNA-HuR delivery affects survival and oxidative stress markers under temperature and excitotoxic insults. In vivo approach consisted of two different settings. In first one, 35 eight-week-old rats received intravitreal injection of AAV-shRNA-HuR or AAV-shRNA scramble control. Animals underwent electroretinography tests and were sacrificed 2, 4 or 6 months after injection. Retinas and optic nerves were collected and processed for immunostainings, electron microscopy and stereology. For the second approach, animals received similar gene constructs. To induce chronic glaucoma, 8 weeks after AAV injection, unilateral episcleral vein cauterization was performed. Animals from each group received intravitreal injection of metallothionein II. Animals underwent electroretinography tests and were sacrificed 8 weeks later. Retinas and optic nerves were collected and processed for immunostainings, electron microscopy and stereology.
UNASSIGNED: Silencing of hur induced apoptosis and increased oxidative stress markers in B-35 cells. Additionally, shRNA treatment impaired the cellular stress response to temperature and excitotoxic insults. In vivo, RGC count was decreased by 39% in shRNA-HuR group 6 months after injection, when compared to shRNA scramble control group. In neuroprotection study, the average loss of RGCs was 35% in animals with glaucoma treated with metallothionein and shRNA-HuR and 11.4% in animals with glaucoma treated with metallothionein and the scramble control shRNA. An alteration in HuR cellular content resulted in diminished photopic negative responses in the electroretinogram.
UNASSIGNED: Based on our findings, we conclude that HuR is essential for the survival and efficient neuroprotection of RGC and that the induced alteration in HuR content accelerates both the age-related and glaucoma-induced decline in RGC number and function, further confirming HuR\'s key role in maintaining cell homeostasis and its possible involvement in the pathogenesis of glaucoma.
摘要：
未经批准：ELAVL1/HuR是转录后水平基因表达的关键调控因子，包括应激反应和稳态维持。这项研究的目的是评估hur沉默对视网膜神经节细胞（RGC）的年龄相关性变性的影响，这可能描述了内源性神经保护机制的效率，以及评估Hur沉默RGC在大鼠青光眼模型中的外源性神经保护能力。
UNASSIGNED：该研究包括体外和体内方法。体外，我们用大鼠B-35细胞进行研究,在温度和兴奋性毒性损伤下，AAV-shRNA-HuR递送是否会影响存活和氧化应激标志物。体内方法由两种不同的设置组成。在第一个，35只8周龄大鼠接受AAV-shRNA-HuR或AAV-shRNA扰乱对照的玻璃体内注射。动物接受视网膜电图测试，并在注射后2、4或6个月处死。收集视网膜和视神经并进行免疫染色处理，电子显微镜和体视学。对于第二种方法，动物接受了类似的基因构建体。诱发慢性青光眼，AAV注射后8周，进行单侧巩膜上静脉烧灼术.每组动物接受玻璃体内注射金属硫蛋白II。动物进行视网膜电图测试并在8周后处死。收集视网膜和视神经并进行免疫染色处理，电子显微镜和体视学。
UNASSIGNED：沉默hur诱导B-35细胞凋亡和氧化应激标志物增加。此外,shRNA处理损害了细胞对温度和兴奋性毒性损伤的应激反应。在体内,注射后6个月，shRNA-HuR组RGC计数减少39%，与shRNA扰乱对照组相比。在神经保护研究中，在用金属硫蛋白和shRNA-HuR治疗的青光眼动物中,RGC的平均损失为35%,在用金属硫蛋白和scramble对照shRNA治疗的青光眼动物中,RGC的平均损失为11.4%.HuR细胞含量的改变导致视网膜电图中的明视负反应减少。
未经评估：根据我们的发现，我们得出的结论是，HuR对于RGC的存活和有效的神经保护至关重要，并且诱导的HuR含量改变加速了年龄相关和青光眼诱导的RGC数量和功能下降，进一步证实了HuR在维持细胞稳态中的关键作用及其可能参与青光眼的发病机制。
UNASSIGNED：该研究包括体外和体内方法。体外，我们用大鼠B-35细胞进行研究,在温度和兴奋性毒性损伤下，AAV-shRNA-HuR递送是否会影响存活和氧化应激标志物。体内方法由两种不同的设置组成。在第一个，35只8周龄大鼠接受AAV-shRNA-HuR或AAV-shRNA扰乱对照的玻璃体内注射。动物接受视网膜电图测试，并在注射后2、4或6个月处死。收集视网膜和视神经并进行免疫染色处理，电子显微镜和体视学。对于第二种方法，动物接受了类似的基因构建体。诱发慢性青光眼，AAV注射后8周，进行单侧巩膜上静脉烧灼术.每组动物接受玻璃体内注射金属硫蛋白II。动物进行视网膜电图测试并在8周后处死。收集视网膜和视神经并进行免疫染色处理，电子显微镜和体视学。
UNASSIGNED：沉默hur诱导B-35细胞凋亡和氧化应激标志物增加。此外,shRNA处理损害了细胞对温度和兴奋性毒性损伤的应激反应。在体内,注射后6个月，shRNA-HuR组RGC计数减少39%，与shRNA扰乱对照组相比。在神经保护研究中，在用金属硫蛋白和shRNA-HuR治疗的青光眼动物中,RGC的平均损失为35%,在用金属硫蛋白和scramble对照shRNA治疗的青光眼动物中,RGC的平均损失为11.4%.HuR细胞含量的改变导致视网膜电图中的明视负反应减少。
未经评估：根据我们的发现，我们得出的结论是，HuR对于RGC的存活和有效的神经保护至关重要，并且诱导的HuR含量改变加速了年龄相关和青光眼诱导的RGC数量和功能下降，进一步证实了HuR在维持细胞稳态中的关键作用及其可能参与青光眼的发病机制。
