Upregulation of - inhibits high-glucose induced inflammation and oxidative stress in HRECs and attenuates diabetic retinopathy by modulating / signaling pathway: an in vitro study.

PURPOSE

The aim of this study was to investigate the underlying mechanisms of diabetic retinopathy (DR) development.

METHODS

Real-Time qPCR was used to detect - (-) and - () mRNA levels. Western Blot was employed to detect protein levels. Malondialdehyde (MDA) assay kit, superoxide dismutase (SOD) kit and glutathione peroxidase (GSH-Px) kit were used to evaluate oxidative stress in high-glucose treated human retinal endothelial cells (HRECs). Calcein-AM/propidium iodide (PI) double stain kit was employed to detect cell apoptosis. Enzyme-linked ImmunoSorbent Assay (ELISA) was used to detect inflammation associated cytokines secretion. Co-immunoprecipitation (CO-IP) was performed to investigate the interactions between - and . Luciferase reporter gene system was used to detect the transcriptional activity of .

RESULTS

• was significantly downregulated in either DR tissues or high-glucose treated HRECs comparing to the Control groups. Besides, high-glucose (25 mM) inhibited HRECs viability and induced oxidative stress, inflammation associated cytokines (TNF-α, IL-6 and IL-1β) secretion and cell apoptosis, which were all reversed by synergistically overexpressing - and aggravated by knocking down -. Of note, we found that overexpressed - activated / signaling pathway and increased its downstream targets including -, -, and in high-glucose treated HRECs. Further results also showed that - regulated cell viability, oxidative stress, inflammation and apoptosis in high-glucose treated HRECs by activating / signaling pathway.

CONCLUSION

We concluded that overexpressed - alleviated DR progression by activating / signaling pathway.