实验性视神经变性发展过程中视网膜色素上皮细胞中酪氨酸酶表达水平的变化

Aigul M. Barieva, Viktor V. Valiullin, Alexander N. Samoilov
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引用次数: 0

摘要

背景:在各种原因引起的视神经变性中,视网膜神经元是最脆弱的,人们已经对它们在这种情况下的状态进行了足够详细的研究。同时,视网膜色素上皮起着非常重要的作用,它能实现黑色素的合成,并发挥抗氧化、吞噬、运输、保护和平衡功能。酪氨酸酶是该上皮细胞生成黑色素的关键酶。视神经萎缩的功能性视力损害比结构性视力损害出现得晚,而且是在疾病的晚期阶段,因此开发早期诊断视神经萎缩的方法极为重要。视网膜色素上皮对视神经变性过程不可能不敏感。因此,有必要对视网膜色素上皮在视神经变性实验中的变化进行研究,以便更全面地了解在这种情况下视觉分析器的变化过程。目的:根据形态学数据研究视网膜色素上皮细胞在实验性视神经变性不同发育阶段酪氨酸酶表达水平的变化。材料与方法:研究对象是用甲醇建立视神经萎缩模型后的 40 只大鼠眼睛。动物被分为五组:第一组--对照组;第二组至第五组包括分别在建立氨茶碱-甲醇视神经萎缩模型 1、3、6 和 9 周后进行眼球摘除的动物。使用酪氨酸酶酪氨酸酶小鼠单克隆荧光抗体对视网膜色素上皮进行研究。对固定的眼球横切面进行免疫荧光染色。使用 R 4.2.2 统计计算环境对所得数据进行统计分析和可视化。结果:对所得结果的比较分析表明,第五组的酪氨酸酶表达水平平均为 4.21 倍[95% 置信区间(CI)2.49-7.13,P=0.00076],9.31倍(95% CI 5.47-15.85,p=0.00033)、2.63倍(95% CI 1.51-4.58,p=0.0009)和3.44倍(95% CI 1.96-6.03,p=0.00055)。据统计,第一组、第三组和第四组实验样本中的酪氨酸酶水平与第二组相比分别平均高出 2.21 倍(95% CI 1.33-3.66,p=0.0003)、3.53 倍(95% CI 2.08-6.02,p=0.00069)和 2.71 倍(95% CI 1.58-4.65,p=0.00022)。结论:在这些条件下,视神经纤维发生变性过程,视网膜色素上皮的完整性受到破坏,其特征是实验动物视网膜色素上皮中的酪氨酸酶抗体增加。
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Changes in the level of tyrosinase expression in the retinal pigment epithelium during the development of experimental optic nerve degeneration
BACKGROUND: With optic nerve degeneration of various origins, retinal neurons are the most vulnerable, and their state under these conditions has been studied in sufficient detail. At the same time, the retinal pigment epithelium plays a very important role, realizing the synthesis of melanin and performing antioxidant, phagocytic, transport, protective, and homeostatic functions. Tyrosinase serves as a key enzyme in melanogenesis by cells of this epithelium. The development of methods for early diagnosis of optic nerve atrophy is extremely important due to the fact that functional visual impairments appear later than structural ones, at more advanced stages of the disease. The retinal pigment epithelium cannot be insensitive to the processes that occur during optic nerve degeneration. That is why its study in experimental degeneration of the optic nerve is necessary for a more complete understanding of the processes occurring in the visual analyzer under these conditions. AIM: To study changes in the level of tyrosinase expression in the retinal pigment epithelium at different development stages of experimental optic nerve degeneration based on morphological data. MATERIAL AND METHODS: The subjects of the study were 40 rat eyes after creating a model of optic nerve atrophy using methanol. The animals were divided into five groups: the first group — control; the second to fifth groups included animals whose eyes were enucleated 1, 3, 6 and 9 weeks after the creation of the aminophylline-methanol model of optic nerve atrophy, respectively. The retinal pigment epithelium was studied using fluorescent antibodies to tyrosinase Tyrosinase Mouse Monoclonal. Immunofluorescence staining was performed on fixed transverse sections of the eyeball. Statistical analysis and visualization of the obtained data were carried out using the environment for statistical computing R 4.2.2. RESULTS: A comparative analysis of the results obtained revealed that the level of tyrosinase expression in the fifth group was on average 4.21 times [95% confidence interval (CI) 2.49–7.13, p=0.00076], 9.31 times (95% CI 5.47–15.85, p=0.00033), 2.63 times (95% CI 1.51–4.58, p=0.0009) and 3.44 times (95% CI 1.96–6.03, p=0.00055) higher compared to the first, second, third and fourth groups, respectively. The level of tyrosinase in experimental samples in the first, third and fourth groups was statistically higher compared to the level of tyrosinase in the second group by an average of 2.21 times (95% CI 1.33–3.66, p=0.0003), 3.53 times (95% CI 2.08–6.02, p=0.00069) and 2.71 times (95% CI 1.58–4.65, p=0.00022), respectively. CONCLUSION: As degenerative processes develop in the optic nerve fibers under these conditions, the integrity of the retinal pigment epithelium is disrupted, which is characterized by an increase in antibodies to tyrosinase in the retinal pigment epithelium of experimental animals.
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