Related Experiment Video
Updated: Sep 4, 2025
Author Spotlight: Understanding Retinal Vessel Resilience and Disease Progression
Published on: January 12, 2024
Down-regulation of LCN2 attenuates retinal vascular dysfunction and caspase-1-mediated pyroptosis in diabetes
Xingjie Su1, Pingping Zhou1, Yanxiu Qi1
1Department of Ophthalmology, The First Affiliated Hospital of Jiamusi University, Jiamusi, China.
Background:
Diabetic retinopathy (DR) is a diabetic microangiopathy with increasing incidence, which seriously threatens the quality of life of patients. This study investigated the molecular regulation mechanism of lipocalin-2 (LCN2) in DR by targeting the function of human retinal vascular endothelial cells (HRVECs).
Methods:
The expression of LCN2 in the retinal tissue of diabetic and high glucose (HG)-induced HRVECs was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis and western blotting assay. After intravitreal injection of adeno-associated virus (AAV)-NC or AAV-sh-LCN2, in vivo experiments, hematoxylin and eosin (H&E) staining, and retinal trypsin digestion experiments were performed to analyze the effect of LCN2 silencing on DR retinal tissue. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining was used to evaluate apoptosis and immunohistochemical (IHC) staining was performed to detect the expressions of caspase-1. Western blot was used to detect the expressions of pyroptosis-associated proteins. After transfection of sh-NC and sh-LCN2, the function of HRVECs cells induced by HG was evaluated by wound healing assay, Transwell assay, and tube formation assay.
Results:
The expression of LCN2 was significantly up-regulated in diabetic retinal tissue and HG-induced HRVECs. In vivo experiments showed that LCN2 silencing can significantly reduce diabetic retinal injury. Cell function experiments also revealed that LCN2 silencing inhibited cell migration, invasion, and angiogenesis. Flow cytometry and immunofluorescence staining showed that downregulation of LCN2 could inhibit caspase-1 mediated pyroptosis in HG-induced HRVECs.
Conclusions:
Down-regulation of LCN2 can significantly inhibit cell migration, invasion, and angiopoiesis, and pyroptosis regulated by caspase-1, thus attenuating the progression of DR.
Insights
Lipocalin-2 (LCN2) is upregulated in diabetic retinopathy (DR). Silencing LCN2 reduces retinal injury, inhibits cell migration and angiogenesis, and attenuates pyroptosis, thereby slowing DR progression.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Diabetic retinopathy (DR) is a growing microangiopathy impacting patient quality of life.
- Lipocalin-2 (LCN2) is investigated for its molecular regulatory role in DR.
- Human retinal vascular endothelial cells (HRVECs) are targeted to understand LCN2's function.
Purpose of the Study:
- To investigate the molecular mechanism of LCN2 in diabetic retinopathy.
- To determine the effect of LCN2 on human retinal vascular endothelial cells (HRVECs).
Main Methods:
- Detected LCN2 expression in diabetic retinal tissue and high glucose (HG)-induced HRVECs using RT-qPCR and Western blotting.
- Utilized intravitreal AAV injection for in vivo LCN2 silencing, followed by H&E and retinal trypsin digestion.
- Assessed cell migration, invasion, angiogenesis, apoptosis (TUNEL), and pyroptosis (caspase-1, Western blot) in HG-induced HRVECs.
Main Results:
- LCN2 expression was significantly upregulated in diabetic retinas and HG-induced HRVECs.
- LCN2 silencing reduced diabetic retinal injury, inhibited cell migration, invasion, and angiogenesis.
- Downregulation of LCN2 inhibited caspase-1 mediated pyroptosis in HG-induced HRVECs.
Conclusions:
- LCN2 downregulation attenuates diabetic retinopathy progression.
- Inhibiting LCN2 reduces cell migration, invasion, angiogenesis, and caspase-1 regulated pyroptosis.
- LCN2 plays a key role in the molecular mechanisms underlying diabetic retinopathy.
