Retinal ganglion cell neuroprotection induced by activation of alpha7 nicotinic acetylcholine receptors.
Retinal ganglion cell neuroprotection induced by activation of alpha7 nicotinic acetylcholine receptors.
复制标题
视网膜神经节细胞神经保护通过α7烟碱乙酰胆碱受体的激活诱导。
DOI：
10.1016/j.neuropharm.2015.07.036
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发表时间：
2015-12
影响因子：
4.7
通讯作者：
Linn CL
中科院分区：
文献类型：
--
作者：
Mata D;Linn DM;Linn CL
在体内青光眼模型中，当将α 7 nAChR激动剂PNU-282987注射到成年Long Evans大鼠眼的玻璃体腔中时，该激动剂先前已显示出对视网膜神经节细胞（RGC）损失的神经保护作用。在这里，我们表征了PNU-282987在神经纤维和视网膜神经节细胞层的神经保护作用，确定了当激动剂作为滴眼剂施用时发生神经保护，并使用LC/MS/MS验证了视网膜中激动剂的检测。将高渗盐水注射到巩膜外静脉中以诱导瘢痕组织并增加眼内压。在一个月内，与未处理的条件相比，该程序产生了RGC的显著损失。用抗Thy 1.1抗体免疫染色后对RGC进行定量，并使用共聚焦显微镜进行成像。在剂量反应研究中，每天两次将PNU-282987浓度应用于动物右眼，而左眼作为内部对照。PNU-282987滴眼液使用100 µM至2 mM PNU-282987浓度以剂量依赖性方式对RGC损失产生神经保护作用。LC/MS/MS结果表明，当作为滴眼剂施用时，在视网膜中检测到PNU-282987，在血浆中测得相对少量的PNU-282987，在心脏组织中未检测到PNU-282987。这些结果支持PNU-282987滴眼液应用可预防与青光眼相关的RGC损失的假设，这可导致涉及α 7 nAChR的疾病的神经保护性治疗。
The α7nAChR agonist, PNU-282987, has previously been shown to have a neuroprotective effect against loss of retinal ganglion cells (RGCs) in an in vivo glaucoma model when the agent was injected into the vitreous chamber of adult Long Evans rat eyes. Here, we characterized the neuroprotective effect of PNU-282987 at the nerve fiber and retinal ganglion cell layer, determined that neuroprotection occurred when the agonist was applied as eye drops and verified detection of the agonist in the retina, using LC/MS/MS. To induce glaucoma-like conditions in adult Long Evans rats, hypertonic saline was injected into the episcleral veins to induce scar tissue and increase intraocular pressure. Within one month, this procedure produced significant loss of RGCs compared to untreated conditions. RGCs were quantified after immunostaining with an antibody against Thy 1.1 and imaged using a confocal microscope. In dose-response studies, concentrations of PNU-282987 were applied to the animal’s right eye two times each day, while the left eye acted as an internal control. Eye drops of PNU-282987 resulted in neuroprotection against RGC loss in a dose-dependent manner using concentrations between 100 µM and 2 mM PNU-282987. LC/MS/MS results demonstrated that PNU-282987 was detected in the retina when applied as eye drops, relatively small amounts of PNU-282987 were measured in blood plasma and no PNU-282987 was detected in cardiac tissue. These results support the hypothesis that eye drop application of PNU-282987 can prevent loss of RGCs associated with glaucoma, which can lead to neuroprotective treatments for diseases that involve α7nAChRs.