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糖尿病相关刺激下的视网膜色素上皮细胞中 AlphaB- 晶状体蛋白的下调

Downregulation of AlphaB-crystallin in Retinal Pigment Epithelial Cells Exposed to Diabetes-related Stimuli and .

机构信息

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.

出版信息

In Vivo. 2022 Jan-Feb;36(1):132-139. doi: 10.21873/invivo.12684.

Abstract

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.

MATERIALS AND METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

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。

材料和方法

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。

结果

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。

结论

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。

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