新型Nrf2诱导剂化合物ITH12674减缓小鼠rd10模型中视网膜色素变性的进展。

New Nrf2-Inducer Compound ITH12674 Slows the Progression of Retinitis Pigmentosa in the Mouse Model rd10.

作者信息

Campello Laura, Kutsyr Oksana, Noailles Agustina, Michalska Patrycja, Fernández-Sánchez Laura, Martínez-Gil Natalia, Ortuño-Lizarán Isabel, Sánchez-Sáez Xavier, de Juan Emilio, Lax Pedro, León Rafael, García Antonio G, Cuenca Nicolás, Maneu Victoria

机构信息

Departamento de Fisiología, Genética y Microbiología, Universidad de Alicante, Alicante, Spain.

Neurobiology-Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.

出版信息

Cell Physiol Biochem. 2020 Feb 7;54(1):142-159. doi: 10.33594/000000210.

DOI:10.33594/000000210

PMID:32028545

Abstract

BACKGROUND/AIMS: It is well established that oxidative stress and inflammation are common pathogenic features of retinal degenerative diseases. ITH12674 is a novel compound that induces the transcription factor Nrf2; in so doing, the molecule exhibits anti-inflammatory, and antioxidant properties, and affords neuroprotection in rat cortical neurons subjected to oxidative stress. We here tested the hypothesis that ITH12674 could slow the retinal degeneration that causes blindness in rd10 mice, a model of retinitis pigmentosa.

METHODS

Animals were intraperitoneally treated with 1 or 10 mg/Kg ITH12674 or placebo from P16 to P30. At P30, retinal functionality and visual acuity were analyzed by electroretinography and optomotor test. By immunohistochemistry we quantified the photoreceptor rows and analyzed their morphology and connectivity. Oxidative stress and inflammatory state was studied by Western blot, and microglia reactivity was monitored by flow cytometry. The blood-brain barrier permeation of ITH12674 was evaluated using a PAMPA-BBB assay.

RESULTS

In rd10 mice treated with 10 mg/Kg of the compound, the following changes were observed (with respect to placebo): (i) a decrease of vision loss with higher scotopic a- and b-waves; (ii) increased visual acuity; (iii) preservation of cone photoreceptors morphology, as well as their synaptic connectivity; (iv) reduced expression of TNF-α and NF-κB; (v) increased expression of p38 MAPK and Atg12-Atg5 complex; and (vi) decreased CD11c, MHC class II and CD169 positive cell populations.

CONCLUSION

These data support the view that a Nrf2 inducer compound may arise as a new therapeutic strategy to combat retinal neurodegeneration. At present, we are chemically optimising compound ITH12674 with the focus on improving its neuroprotective potential in retinal neurodegenerative diseases.

摘要

背景/目的：氧化应激和炎症是视网膜退行性疾病常见的致病特征，这一点已得到充分证实。ITH12674是一种新型化合物，可诱导转录因子Nrf2；在此过程中，该分子表现出抗炎和抗氧化特性，并为遭受氧化应激的大鼠皮质神经元提供神经保护。我们在此检验了ITH12674可减缓导致rd10小鼠失明的视网膜退化这一假设，rd10小鼠是色素性视网膜炎模型。

方法

从出生后第16天（P16）至第30天（P30），对动物腹腔注射1或10 mg/Kg的ITH12674或安慰剂。在P30时，通过视网膜电图和视动试验分析视网膜功能和视力。通过免疫组织化学对光感受器排进行定量，并分析其形态和连接性。通过蛋白质印迹法研究氧化应激和炎症状态，并通过流式细胞术监测小胶质细胞反应性。使用PAMPA - BBB测定法评估ITH12674的血脑屏障通透性。

结果

在用10 mg/Kg该化合物治疗的rd10小鼠中，观察到以下变化（相对于安慰剂）：（i）暗视a波和b波较高，视力丧失减少；（ii）视力提高；（iii）视锥光感受器形态及其突触连接性得以保留；（iv）肿瘤坏死因子-α（TNF-α）和核因子-κB（NF-κB）表达降低；（v）p38丝裂原活化蛋白激酶（p38 MAPK）和自噬相关蛋白12 - 自噬相关蛋白5复合物（Atg12 - Atg5 complex）表达增加；（vi）CD11c、主要组织相容性复合体II类分子（MHC class II）和CD169阳性细胞群体减少。