NLRP3 Blockade Suppresses Pro-Inflammatory and Pro-Angiogenic Cytokine Secretion in Diabetic Retinopathy

Abstract

Background

Inflammation and angiogenesis are the two dominant mechanisms of diabetic retinopathy (DR), which act more as mutual pathways rather than individual processes. However, the underlying mechanism of their interactions is still unclear. Here, we explored the potential crossing points between these pathways and the targeted therapeutic method in rats with DR.

Materials and Methods

Sprague–Dawley rats were randomly assigned to four groups: normal control group, streptozocin (STZ)-induced diabetes mellitus (DM) group, DM+shNC (non-specific negative control shRNA) group, and DM+shNLRP3 group. Silencing the NLR family pyrin domain containing 3 (NLRP3) protein was performed by intravitreal injections of NLRP3-targeted shRNA (shNLRP3) for rats in the DM+shNLRP3 group. All the rats’ retinas were collected for further morphological examination and pro-inflammatory and pro-angiogenic cytokine detection. Human retinal endothelial cells (HRECs) were also employed to explore the underlying mechanism.

Results

NLRP3-targeted shRNA given by intravitreal injection effectively alleviated the retinal histopathological changes in STZ-induced diabetic rats, which reduced the activation of the NLRP3 inflammasome and suppressed the expressions of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and inflammatory cytokines in diabetic rats’ retinas. In HRECs, NLRP3 over-expressing plasmid evoked an increase in pro-inflammatory cytokines and VEGF. In addition, YC-1, a HIF-1α inhibitor, could reverse the NLRP3 over-expression-induced VEGF production but not the pro-inflammatory cytokine expressions.

Conclusion

Our results suggest NLRP3 inflammasome as the potential cross-point between inflammation and pro-angiogenesis in DR and support the effectiveness of NLRP3-targeted shRNA administrated by intravitreal injection in animal models of DR. The protective effect of NLRP3-targeted shRNA may stem from the inhibition of both pro-inflammatory cytokines and HIF-1α/VEGF axis.

Keywords:

• NLRP3 inflammasome
• inflammation
• angiogenesis
• VEGF
• diabetic retinopathy

Acknowledgment

This work was sponsored by the National Natural Science Foundation of China [grant number 81200718 and 81570866].

Disclosure

The authors report no conflicts of interest for this work.