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MiR-200c-3p regulates pyroptosis by targeting SLC30A7 in diabetic retinopathy
Weina Li1, Sheng Yang1, Guangsheng Chen1
1Department of Glaucoma and Cataract, Liuzhou Aier Eye Hospital, Affiliated Hospital of Aier Ophthalmology College of Central South University, Liuzhou, China.
Clinical Relevance:
MicroRNAs (miRNAs) have been reported to be involved in the progression of various diseases. Studying the regulatory mechanisms of miRNAs can help clinical treatment.
Background:
Diabetic retinopathy (DR) is one of the complications of diabetes. The objective of this study was to elucidate the underlying molecular mechanisms by which miR-200c-3p regulates the pyroptosis of DR cell.
Methods:
Human retinal microvascular endothelial cells (HRMECs) and high glucose (HG) cultures established DR cell model in vitro. RT-qPCR is used to detect the expression level of miRNAs. CCK-8 assays and flow cytometry are used to detect apoptosis of HRMECs cell. Western blotting is used to detect cleaved caspase-3, cleaved caspase-1, and N-GSDMD proteins levels in HRMECs. The ELISA assay is used to detect the expression of IL-1β and IL-18. Predict and validate potential binding sites between miR-200c-3p and SLC30A7 by dual luciferase reporter gene analysis.
Results:
The results showed that HG caused damage to HRMECs through the pyroptosis pathway rather than the apoptosis pathway. MiR-200c-3p is highly expressed in HG induced-HRMECs, and knockdown of miR-200c-3p mitigates HG-induced HRMECs pyroptosis. MiR-200c-3p negatively targets SLC30A7 in HRMECs, and miR-200c-3p regulates pyroptosis of HG-induced HRMECs by targeting SLC30A7.
Conclusion:
The results suggest that miR-200c-3p might be a promising interference target for DR prevention and treatment. The results of current study may provide new insights into development of therapeutic strategies for DR.
Insights
MicroRNA miR-200c-3p promotes pyroptosis in diabetic retinopathy (DR) by targeting SLC30A7. Inhibiting miR-200c-3p may offer a new therapeutic strategy for DR treatment.
Area of Science:
- Molecular biology
- Cell biology
- Ophthalmology
Background:
- Diabetic retinopathy (DR) is a diabetes complication.
- Understanding miRNA regulatory mechanisms is crucial for disease treatment.
- This study investigates miR-200c-3p's role in DR cell pyroptosis.
Purpose of the Study:
- To elucidate the molecular mechanisms of miR-200c-3p in regulating pyroptosis in diabetic retinopathy (DR) cells.
- To investigate the potential of miR-200c-3p as a therapeutic target for DR.
Main Methods:
- Established a DR cell model using human retinal microvascular endothelial cells (HRMECs) and high glucose (HG).
- Quantified miRNA expression using RT-qPCR.
- Assessed cell apoptosis and pyroptosis markers (caspase-3, caspase-1, N-GSDMD, IL-1β, IL-18) via flow cytometry, Western blotting, and ELISA.
- Validated miR-200c-3p and SLC30A7 interaction using dual luciferase reporter assays.
Main Results:
- High glucose (HG) induced pyroptosis, not apoptosis, in HRMECs.
- miR-200c-3p expression was elevated in HG-induced HRMECs.
- Knockdown of miR-200c-3p reduced HG-induced pyroptosis.
- miR-200c-3p directly targets SLC30A7, regulating HG-induced pyroptosis.
Conclusions:
- miR-200c-3p promotes pyroptosis in DR by targeting SLC30A7.
- miR-200c-3p represents a potential therapeutic target for DR.
- Findings offer new insights for developing DR treatment strategies.
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