Chistyakov Dmitry V, Tiulina Veronika V, Gancharova Olga S, Baksheeva Viktoriia E, Goriainov Sergei V, Shebardina Natalia G, Ivlev Vasily A, Komarov Sergey V, Shevelyova Marina P, Tikhomirova Natalia K, Philippov Pavel P, Vasil'ev Vasiliy G, Sergeeva Marina G, Permyakov Sergei E, Iomdina Elena N, Tsvetkov Philipp O, Senin Ivan I, Zernii Evgeni Yu
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia.
Pharmacy Resource Center, Peoples' Friendship University of Russia (RUDN University), 117198 Moscow, Russia.
Int J Mol Sci. 2024 Nov 30;25(23):12910. doi: 10.3390/ijms252312910.
Autoimmune uveitis is a relapsing blind-causing ocular condition with complex pathogenesis that is not completely understood. There is a high demand for accurate animal models of experimental autoimmune uveitis (EAU) suitable for elucidating the molecular mechanisms of the disease and testing new therapeutic approaches. Here, we demonstrated that photoreceptor Ca/Zn-sensor protein recoverin is a uveoretinal antigen in albino rabbits provoking typical autoimmune chorioretinitis 2-4 weeks after immunization. The pathologic process in recoverin-induced EAU shared features with human disease and included lymphocytic infiltration of the retina, Dalen-Fuchs nodules and foci of subtotal or total retinal atrophy, manifested as a decrease in amplitude of the a-wave of the electroretinogram. In some cases, changes in the retinal vascular pattern and subretinal hemorrhages were also observed. These signs were accompanied by a gradual accumulation of serum antibodies against recoverin. Biochemical examination of the aqueous humor (AH) revealed typical characteristics of inflammation and oxidative stress, including increased levels of TNF-α and IL-6 and decreased levels of IL-10, as well as decreased total antioxidant activity, superoxide dismutase and glutathione peroxidase activities, and increased zinc concentration. Consistently, metabolomic and targeted lipidomic analysis of AH showed high lactate and low ascorbic acid levels in early EAU; increased levels of key pro-inflammatory signaling lipids such as PGE2, TXB2, 11-HETE and Lyso-PAF; and reduced levels of the anti-inflammatory fatty acid DHA in advanced stages of the disease. Uveitic AH became enriched with recoverin, confirming disruption of the blood-ocular barrier and photoreceptor damage. Notably, the application of mitochondria-targeted antioxidant therapy impeded EAU progression by maintaining local antioxidant activity and suppressing TNF-α, IL-6 and PGE2 signaling. Overall, our results demonstrate that recoverin-induced EAU in rabbits represents an accurate model of human autoimmune posterior uveitis and suggest new directions for its therapy that can be trialed using the developed model.
自身免疫性葡萄膜炎是一种导致失明的复发性眼部疾病，其发病机制复杂，尚未完全明确。对于适用于阐明该疾病分子机制和测试新治疗方法的准确实验性自身免疫性葡萄膜炎（EAU）动物模型有很高的需求。在此，我们证明光感受器钙/锌传感器蛋白恢复蛋白是白化兔中的葡萄膜视网膜抗原，在免疫后2-4周引发典型的自身免疫性脉络膜视网膜炎。恢复蛋白诱导的EAU的病理过程与人类疾病有共同特征，包括视网膜淋巴细胞浸润、达伦-富克斯结节以及视网膜部分或完全萎缩灶，表现为视网膜电图a波振幅降低。在某些情况下，还观察到视网膜血管模式改变和视网膜下出血。这些体征伴随着血清中抗恢复蛋白抗体的逐渐积累。房水（AH）的生化检查显示出炎症和氧化应激的典型特征，包括肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平升高、白细胞介素-10（IL-10）水平降低，以及总抗氧化活性、超氧化物歧化酶和谷胱甘肽过氧化物酶活性降低，锌浓度升高。一致地，AH的代谢组学和靶向脂质组学分析显示，在EAU早期乳酸水平高而抗坏血酸水平低；关键促炎信号脂质如前列腺素E2（PGE2）、血栓素B2（TXB2）、11-羟二十碳四烯酸（11-HETE）和溶血血小板活化因子（Lyso-PAF）水平升高；在疾病晚期抗炎脂肪酸二十二碳六烯酸（DHA）水平降低。葡萄膜炎性AH中恢复蛋白富集，证实血眼屏障破坏和光感受器损伤。值得注意的是，线粒体靶向抗氧化治疗通过维持局部抗氧化活性并抑制TNF-α、IL-6和PGE2信号传导来阻碍EAU进展。总体而言，我们的结果表明，恢复蛋白诱导的兔EAU代表了人类自身免疫性后葡萄膜炎的准确模型，并为其治疗提出了新方向，可使用所建立的模型进行试验。
