Eye Center of the Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, P.R. China.
Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, P.R. China.
In Vivo. 2022 Jan-Feb;36(1):132-139. doi: 10.21873/invivo.12684.
BACKGROUND/AIM: AlphaB-crystallin plays a pivotal role in many diseases. However, the involvement of alphaB-crystallin in retinal pigment epithelial (RPE) cells with diabetes stimuli remains unknown. The aim of this study is to examine the alterations of RPE cells and alphaB-crystallin expression in diabetic models in vivo and in vitro.
Diabetic conditions in mice were induced by streptozotocin (STZ). The thickness of the RPE/choroid complex was measured by optical coherence tomography (OCT). Periodic acid-Schiff (PAS) staining was used to investigate the choriocapillaris in histological sections of murine eyeballs and oxidative stress was evaluated using immunofluorescence with anti-4-hydroxynonenal (HNE) antibody. AlphaB-crystallin expression was examined in the RPE/choroid complex using ELISA. Real-Time PCR was performed to evaluate the alphaB-crystallin expression in cultured human RPE cells with high glucose or following advanced glycation end-products (AGE) stimulation.
In diabetic mice, OCT-based RPE/choroidal layers were thickened 2 months after STZ stimulation, where PAS-positive dilated choriocapillaris was noted. Immunoreactivity of 4-HNE was strongly observed in the RPE layer, from which a significant number of RPE cells was lost. Meanwhile, alphaB-crystallin expression in 2-month STZ mice was significantly lower compared to controls. In accordance with these results, in vitro data showed that the alphaB-crystallin expression was also significantly lower in RPE cells with high glucose or following AGE stimulation compared to untreated cells.
In both types of diabetic models the expression of alphaB-crystallin was found to be downregulated in RPE cells and was associated with increased levels of oxidative stress.
背景/目的:αB-晶状体蛋白在许多疾病中起着关键作用。然而,αB-晶状体蛋白在糖尿病刺激的视网膜色素上皮(RPE)细胞中的作用尚不清楚。本研究旨在研究体内和体外糖尿病模型中 RPE 细胞和 αB-晶状体蛋白表达的变化。
通过链脲佐菌素(STZ)诱导小鼠糖尿病。使用光学相干断层扫描(OCT)测量 RPE/脉络膜复合体的厚度。用过碘酸希夫(PAS)染色法研究小鼠眼球组织切片中的脉络膜毛细血管,并用抗 4-羟壬烯醛(HNE)抗体的免疫荧光法评估氧化应激。使用 ELISA 法检测 RPE/脉络膜复合体中的αB-晶状体蛋白表达。通过实时 PCR 评估高糖或晚期糖基化终产物(AGE)刺激培养的人 RPE 细胞中的αB-晶状体蛋白表达。
在糖尿病小鼠中,STZ 刺激 2 个月后,基于 OCT 的 RPE/脉络膜层增厚,可见 PAS 阳性扩张的脉络膜毛细血管。在 RPE 层中强烈观察到 4-HNE 的免疫反应性,其中大量 RPE 细胞丢失。同时,与对照组相比,2 个月 STZ 小鼠的αB-晶状体蛋白表达明显降低。与这些结果一致,体外数据显示,与未处理的细胞相比,高糖或 AGE 刺激后的 RPE 细胞中αB-晶状体蛋白的表达也明显降低。
在这两种类型的糖尿病模型中,RPE 细胞中αB-晶状体蛋白的表达均下调,并与氧化应激水平升高相关。