成年色素性小鼠体内局部发光二极管诱导的锥形光感受器光毒性模型：碱性成纤维细胞生长因子的保护作用。

An in vivo model of focal light emitting diode-induced cone photoreceptor phototoxicity in adult pigmented mice: Protection with bFGF.

机构信息

Departamento de Oftalmología, Universidad de Murcia e Instituto Murciano de Investigación Biosanitaria (IMIB) Virgen de la Arrixaca. Campus de CC de la Salud, 30120, El Palmar, Murcia, Spain.

出版信息

Exp Eye Res. 2021 Oct;211:108746. doi: 10.1016/j.exer.2021.108746. Epub 2021 Aug 24.

DOI:10.1016/j.exer.2021.108746

PMID:34450185

Abstract

PURPOSE

To develop a model of focal injury by blue light-emitting diode (LED)-induced phototoxicity (LIP) in pigmented mouse retinas and to study the effects on cone, Iba-1 cells and retinal pigment epithelium (RPE) cell populations after administration of basic fibroblast growth factor (bFGF) and minocycline, alone or combined.

METHODS

In anesthetized dark-adapted adult female pigmented C57BL/6 mice, left pupils were dilated and the eye exposed to LIP (500 lux, 45 s). The retina was monitored longitudinally in vivo with SD-OCT for 7 days (d). Ex vivo, the effects of LIP and its protection with bFGF (0.5 μg) administered alone or combined with minocycline (45 mg/kg) were studied in immunolabeled arrestin-cone outer segments (aOS) and quantified within a predetermined fixed-size circular area (PCA) centered on the lesion in flattened retinas at 1, 3, 5 or 7d. Moreover, Iba-1 cells and RPE cell morphology were analysed with Iba-1 and ZO-1 antibodies, respectively.

RESULTS

LIP caused a focal lesion within the superior-temporal retina with retinal thinning, particularly the outer retinal layers (116.5 ± 2.9 μm to 36.8 ± 6.3 μm at 7d), and with progressive diminution of aOS within the PCA reaching minimum values at 7d (6218 ± 342 to 3966 ± 311). Administration of bFGF alone (4519 ± 320) or in combination with minocycline (4882 ± 446) had a significant effect on aOS survival at 7d and Iba-1 cell activation was attenuated in the groups treated with minocycline. In parallel, the RPE cell integrity was progressively altered after LIP and administration of neuroprotective components had no restorative effect at 7d.

CONCLUSIONS

LIP resulted in progressive outer retinal damage affecting the OS cone population and RPE. Administration of bFGF increased aOS survival but did not prevent RPE deterioration.

摘要

目的

建立蓝光发光二极管（LED）诱导光毒性（LIP）致色素鼠视网膜局灶损伤模型，并研究碱性成纤维细胞生长因子（bFGF）和米诺环素单独或联合应用后对锥体、Iba-1 细胞和视网膜色素上皮（RPE）细胞群体的影响。

方法

在麻醉的暗适应成年雌性色素 C57BL/6 小鼠中，扩瞳并暴露于 LIP（500lux，45s）。用 SD-OCT 在体内对视网膜进行 7 天（d）的纵向监测。在离体研究中，用 LIP 及其用 bFGF（0.5μg）单独或联合米诺环素（45mg/kg）进行保护的作用，并在扁平视网膜中以损伤为中心的预定固定大小圆形区域（PCA）内进行免疫标记的 arrestin-锥体外节（aOS）定量分析，于 1、3、5 或 7d 时进行。此外，用 Iba-1 和 ZO-1 抗体分别分析 Iba-1 细胞和 RPE 细胞形态。

结果

LIP 在视网膜的上颞区引起局灶性病变，导致视网膜变薄，特别是外视网膜层（7d 时为 116.5±2.9μm 至 36.8±6.3μm），PCA 内 aOS 逐渐减少，7d 时达到最小值（6218±342 至 3966±311）。bFGF 单独给药（4519±320）或与米诺环素联合给药（4882±446）对 7d 时 aOS 存活有显著影响，而米诺环素治疗组的 Iba-1 细胞激活受到抑制。同时，LIP 后 RPE 细胞完整性逐渐改变，神经保护成分给药在 7d 时无修复作用。