June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Nr1d1 protects Endothelial Cells against high glucose- and hypoxia-induced injury by promoting autophagy in diabetic retinopathy
Author Affiliations & Notes
  • Suna Ye
    Aier Eye Hospital, Jinan University, Guangzhou, China
    Aier Eye Institute, ChangSha, China
  • Jiansu Chen
    Aier Eye Institute, ChangSha, China
  • Shibo Tang
    Aier Eye Hospital, Jinan University, Guangzhou, China
    Aier Eye Institute, ChangSha, China
  • Footnotes
    Commercial Relationships   Suna Ye None; Jiansu Chen None; Shibo Tang None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1007. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Suna Ye, Jiansu Chen, Shibo Tang; Nr1d1 protects Endothelial Cells against high glucose- and hypoxia-induced injury by promoting autophagy in diabetic retinopathy. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1007.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
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.

 

Fig.1 (A) NR1D1 overexpression. (B) ECs cell viability. (C) Cell migration. (D) Migration analysis. (E-H) Protein expression and quantification. ****p < 0.0001 versus control, ###P<0.001 versus HG-CoCl2.

Fig.1 (A) NR1D1 overexpression. (B) ECs cell viability. (C) Cell migration. (D) Migration analysis. (E-H) Protein expression and quantification. ****p < 0.0001 versus control, ###P<0.001 versus HG-CoCl2.

 

Fig.2 (A, B) Protein expression and quantification of LC3B, and autophagic flux. ****p < 0.0001 versus control, ####P<0.0001 versus HG-CoCl2. (C-E) Apoptosis-associated proteins expression and quantification. *p < 0.05 ****p < 0.0001 versus HG-CoCl2, ####P<0.001 versus OE+HG-CoCl2.

Fig.2 (A, B) Protein expression and quantification of LC3B, and autophagic flux. ****p < 0.0001 versus control, ####P<0.0001 versus HG-CoCl2. (C-E) Apoptosis-associated proteins expression and quantification. *p < 0.05 ****p < 0.0001 versus HG-CoCl2, ####P<0.001 versus OE+HG-CoCl2.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×