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C1q/tumor necrosis factor-related protein-3 (CTRP3) activated by forkhead box O4 (FOXO4) down-regulation protects
XiuYa Zeng1,2, YouYuan Peng3, YanFeng Wang1,2
1Department of Clinical Laboratory, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian, China.
Abstract:
Diabetic retinopathy (DR) remains a major cause of blindness among diabetes mellitus patients. C1q/tumor necrosis factor-related protein-3 (CTRP3) is a novel adipokine which is associated with multiple types of metabolism. Nevertheless, little is known about the role of CTRP3 in high glucose (HG)-induced human retinal pericytes (HRPs). This study set out to assess the influence of CTRP3 on HG-induced HRPs and elucidate the latent regulatory mechanism. RT-qPCR and Western blot were to analyze CTRP3 and forkhead box O4 (FOXO4) expression. Western blot was also utilized to detect the protein levels of apoptosis-related factors and nuclear factor erythroid 2-related factor 2 (Nrf2)/Nuclear factor-kappaB (NF-κB) signaling-related factors. CCK-8 was to measure cell proliferation while TUNEL assay was to estimate cell apoptosis. Levels of oxidative stress biomarkers including manganese (MnSOD), catalase (CAT) and malonedialdehyde (MDA) were evaluated by the corresponding kits. JASPAR database, ChIP and luciferase reporter assay were to verify the interaction between FOXO4 and CTRP3 promoter. The experimental results uncovered that CTRP3 expression was decreased in HG-stimulated HRPs. Moreover, CTRP3 overexpression strengthened the viability while abrogated the apoptosis and oxidative stress of HG-induced HRPs. Furthermore. FOXO4 was up-regulated in HG-induced HRPs. Besides, FOXO4 bond to CTRP3 promoter and inhibited CTRP3 transcription to modulate the Nrf2/NF-κB signaling pathway. FOXO4 up-regulation reversed the influence of CTRP3 elevation on the proliferation, apoptosis and oxidative stress of HG-induced HRPs. To be summarized, CTRP3 negatively modulated by FOXO4 prevented HG-induced oxidative damage in DR via modulation of Nrf2/NF-κB signaling.
Insights
C1q/tumor necrosis factor-related protein-3 (CTRP3) protects against high glucose-induced damage in diabetic retinopathy (DR). Forkhead box O4 (FOXO4) inhibits CTRP3, exacerbating oxidative stress and apoptosis in retinal pericytes.
Area of Science:
- Ophthalmology
- Endocrinology
- Molecular Biology
Background:
- Diabetic retinopathy (DR) is a leading cause of blindness in diabetes mellitus patients.
- C1q/tumor necrosis factor-related protein-3 (CTRP3), a novel adipokine, has roles in metabolism, but its function in DR is unclear.
- High glucose (HG) conditions impact human retinal pericytes (HRPs), crucial cells in retinal vasculature.
Purpose of the Study:
- To investigate the effect of CTRP3 on HG-induced HRPs.
- To elucidate the regulatory mechanism involving CTRP3 in HG-induced HRPs.
- To assess the role of CTRP3 in the context of diabetic retinopathy pathogenesis.
Main Methods:
- Quantitative real-time PCR (RT-qPCR) and Western blot for CTRP3 and FOXO4 expression analysis.
- Cell Counting Kit-8 (CCK-8) for proliferation and TUNEL assay for apoptosis assessment.
- Analysis of oxidative stress biomarkers (MnSOD, CAT, MDA) and verification of FOXO4-CTRP3 interaction using ChIP and luciferase assays.
Main Results:
- CTRP3 expression was reduced, while FOXO4 expression was elevated in HG-stimulated HRPs.
- CTRP3 overexpression improved HRP viability, reduced apoptosis, and mitigated oxidative stress.
- FOXO4 directly binds to the CTRP3 promoter, inhibiting its transcription and modulating the Nrf2/NF-κB signaling pathway.
Conclusions:
- CTRP3 plays a protective role against HG-induced oxidative damage in HRPs, relevant to DR.
- FOXO4 negatively regulates CTRP3, contributing to HG-induced cellular damage in DR.
- CTRP3, via modulation of the Nrf2/NF-κB pathway, offers a potential therapeutic target for DR.
