Abstract
Purpose :
Nuclear receptor subfamily 1 group D member 1 (NR1D1) plays a vital role in the regulation of circadian rhythms, lipid, and glucose metabolism. However, whether NR1D1 mediates the pathogenesis of Diabetic retinopathy (DR) and related mechanisms remains poorly defined. This study determined the effect of NR1D1 on high glucose and hypoxia-treated Human Umbilical Vein Endothelial Cells (ECS) and the underlying mechanisms.
Methods :
ECs were exposed to 5.5 mmol/L of glucose (normal control), and 30 mmol/L of glucose plus 150 mmol/L of CoCl2 (HG-CoCl2). NR1D1 was overexpressed with lentivirus. CCK8, transwell, and TUNEL assays were used to identify the cell viability, migration, and apoptosis potential. Western blot was carried out to examine the levels of proteins associated with apoptosis and autophagy. Chloroquine (CQ) was used to block the autophagic flux.
Results :
Exposed to HG-CoCl2 significantly inhibited cell viability (80%, p<0.0001) but increased migration (p=0.0014). The protein levels of Bax, Cleaved-Caspase3, and Caspase3 were significantly upregulated. Overexpression of NR1D1 effectively relieved HG-CoCl2 -reduced cell viability (100%, p<0.0001), impeded HG-CoCl2-induced migration (p<0.0001), decreased levels of Bax, and cleaved-Caspase3. Besides, HG-CoCl2 significantly suppressed cell autophagy, decreased LC3II expression, and suppressed autophagic flux. NR1D1 overexpression restored the damaged autophagy, increased LC3II expression and autophagic flux. Autophagy inhibitor 3-MA counteracted the improvement from NR1D1 overexpression on HG-CoCl2-induced apoptosis promotion, increased the level of Bax and Cleaved-Caspase 3.
Conclusions :
Our results prove that NR1D1 has a protective effect on HG-CoCl2 -induced injury of ECs, and suggest its therapeutic potential.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.