Abstract
Purpose :
OLT1177 (Dapansutrile) is an effective inhibitor targeted nucleotide-binding domain leucine-rich repeat and pyrin domain containing receptor 3 (NLRP3) inflammasome. Here we aimed to explore therapeutic effect of OLT1177 in oxygen-induced retinopathy (OIR) model and its potential mechanism.
Methods :
OIR model was induced in C57BL/6J newborn mice and OLT1177 was delivered to pups through intraperitoneal injection at P12. Flat mount immunofluorescence and hematoxylin and eosin staining were used to evaluate retinal pathological neovascularization. Western blot and real time PCR were performed to quantify the levels of NLRP3 and inflammatory factors, including IL-1β and TNF-α. Cell counting kit-8 assay was used to evaluate the proliferation of BV2 cells and HUVECs under the hypoxia condition. Meanwhile, EdU incorporation assay, wound-healing assay, transwell assay and matrigel assay are applied to evaluate the proliferation, migration and tube formation of HUVECs.
Results :
OLT1177 administration (100μM) significantly attenuated pathological neovascularization (p<0.01) and reduced 30% of avascular area (P<0.005). Moreover, OLT1177 reduced aggregation and activation of microglia and leakage of retinal vessels. No significant change was found in the expression of NLRP3，whereas the expression of cleaved caspase-1, IL-1β and TNF-α were differentially decreased. Meanwhile, Immunofluorescence analysis showed NLRP3 co-located with both microglia and endothelial cell. OLT1177 inhibited the proliferation of BV2 cells and HUVECs. And the migration and tube formation of hypoxia-induced HUVECs were inhibited after OLT1177 intervention.
Conclusions :
Our study showed that OLT1177 exhibited highly antiangiogenic effects through inhibiting the NLRP3-dependent pathway, suggesting an alternative treatment for RNV.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.