Abstract

Purpose

In the study, we aimed to explore the mechanism of leukaemia inhibitory factor (LIF) affects hyperglycaemic induced retinopathy by regulating CaMKII-CREB pathway.

Methods

Human retinal endothelial cell (HRECs) induced by high glucose to simulate one of the pathogenesis in the diabetic retinopathy (DR) model. After LIF treatment, cell viability was detected by CCK-8 and apoptosis was detected by flow cytometry. Angiogenesis was detected by in vitro tube formation. The expression levels of inflammatory, angiogenesis related proteins and CaMKII-CREB were detected by western blot. The gene level of angiogenesis was detected by qRT-PCR. HE staining was used to detect pathological changes of retinopathy in diabetic mice after LIF treatment.

Results

Our results showed that LIF significantly increased hyperglycaemic-induced cell viability and inhibited apoptosis. Western blot results showed that LIF could down-regulate the expression levels of inflammatory cytokines such as IL-1β, IL-6 and TNF-α. In addition, angiogenesis of HRECs was inhibited by LIF in tubulisation experiments. LIF can down-regulate protein and gene levels of VEGF and HIF-1α via western blot and qRT-PCR. In diabetic mice induced by STZ, LIF could down-regulate the protein level of VEGF, HIF-1α, p-CaMKII and p-CREB, which suggest that LIF could inhibit retinal angiogenesis in diabetic mice. The results of HE staining showed that LIF could alleviate the damage of retinopathy in diabetic mice.

Conclusion

LIF could alleviate the damage of diabetic retinopathy by modulating the CaMKII/CREB signalling pathway to inhibit inflammatory response and angiogenesis.

中文翻译：

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白血病抑制因子对高糖诱导人视网膜内皮细胞功能障碍的保护作用

摘要

目的

本研究旨在探讨白血病抑制因子(LIF)通过调控CaMKII-CREB通路影响高血糖所致视网膜病变的机制。

方法

高糖诱导的人视网膜内皮细胞 (HREC) 模拟糖尿病视网膜病变 (DR) 模型中的一种发病机制。LIF处理后，CCK-8检测细胞活力，流式细胞术检测细胞凋亡。通过体外管形成检测血管生成。免疫印迹检测炎症、血管生成相关蛋白和CaMKII-CREB的表达水平。通过qRT-PCR检测血管生成的基因水平。HE染色用于检测LIF治疗后糖尿病小鼠视网膜病变的病理变化。

结果

我们的结果表明，LIF 显着增加高血糖诱导的细胞活力并抑制细胞凋亡。Western blot结果显示LIF可下调IL-1β、IL-6、TNF-α等炎性细胞因子的表达水平。此外，在管化实验中，LIF 抑制了 HREC 的血管生成。LIF 可以通过蛋白质印迹和 qRT-PCR 下调 VEGF 和 HIF-1α 的蛋白质和基因水平。在 STZ 诱导的糖尿病小鼠中，LIF 可以下调 VEGF、HIF-1α、p-CaMKII 和 p-CREB ​​的蛋白水平，这表明 LIF 可以抑制糖尿病小鼠的视网膜血管生成。HE染色结果表明，LIF可减轻糖尿病小鼠视网膜病变的损伤。

结论

LIF可通过调节CaMKII/CREB信号通路抑制炎症反应和血管生成来减轻糖尿病视网膜病变的损伤。