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Updated: Jun 25, 2025
Comparative Proteomic Analysis of Whole Kidney, Medulla, and Cortical Tubules in Diabetic Pathogenesis of Kidney Injury in Mice
Published on: May 2, 2025
A NOVEL CIRC_SUPT3/MIR-185-5P/G3BP2 CERNA NETWORK REGULATES HIGH GLUCOSE-INDUCED INJURY IN MOUSE PODOCYTE MPC5 CELLS
Yuting Li1, Wenyan Wang2, Na Liu3
1Department of Nephrology, Shanghai TCM-Integrated Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Abstract:
Background: Diabetic nephropathy (DN) is a complication of diabetes that is the leading cause of death in diabetic patients. Circular RNA (circRNA) is a hot topic in the research of human diseases. However, the role of circ_Supt3 in DN remains unclear. Methods: High glucose (HG) treatment of mouse podocyte (MPC5) cells to mimic DN cell injury. Quantitative real-time polymerase chain reaction was performed to detect the expression of circ_Supt3, microRNA-185-5p (miR-185-5p), and GTPase-activating protein-binding protein 2 (G3bp2). 5-Ethynyl-2'-deoxyuridine (EdU) and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium Bromide (MTT) assays were used to examine cell proliferation, and flow cytometry was used to detect cell apoptosis. Western blot was used to assess the levels of relative proteins. Enzyme-linked immunosorbent assay detected the inflammation cytokines. Dual-luciferase reporter and RNA pull-down assays were used to confirm the interaction of miR-185-5p and circ_Supt3 or G3bp2. Results: Circ_Supt3 and G3bp2 were highly expressed and miR-185-5p expression was diminished in DN mice. HG treatment inhibited cell proliferation and accelerated cell apoptosis and inflammation response, and the knockdown of circ_Supt3 reversed these effects. Bioinformatics predicted that circ_Supt3 contained a binding site for miR-185-5p, and G3bp2 was a direct target of miR-185-5p. Circ_Supt3 regulated G3bp2 expression by miR-185-5p. Moreover, the circ_Supt3/miR-185-5p/G3bp2 axis regulated the cell behavior of HG-induced MPC5 cells. Conclusion: Our findings suggest that the knockdown of circ_Supt3 protects mouse MPC5 cells against HG-induced cell injury via the miR-185-5p/G3bp2 axis.
Insights
Knocking down circ_Supt3 protects mouse podocytes from high glucose injury by regulating the miR-185-5p/G3bp2 axis. This study clarifies the role of circ_Supt3 in diabetic nephropathy progression.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- Diabetic nephropathy (DN) is a major cause of mortality in diabetic patients.
- Circular RNAs (circRNAs) are implicated in various human diseases, but their role in DN is not fully understood.
- circ_Supt3's specific function in DN requires elucidation.
Purpose of the Study:
- To investigate the role and mechanism of circ_Supt3 in high glucose-induced mouse podocyte injury.
- To explore the relationship between circ_Supt3, microRNA-185-5p (miR-185-5p), and GTPase-activating protein-binding protein 2 (G3bp2) in DN.
- To determine if the circ_Supt3/miR-185-5p/G3bp2 axis impacts podocyte behavior under diabetic conditions.
Main Methods:
- High glucose (HG) treatment of mouse podocyte (MPC5) cells to simulate DN.
- Quantitative real-time PCR to measure circ_Supt3, miR-185-5p, and G3bp2 expression.
- Cell proliferation (EdU, MTT), apoptosis (flow cytometry), and inflammation assays.
- Western blot for protein analysis; dual-luciferase and RNA pull-down assays for interaction confirmation.
Main Results:
- Circ_Supt3 and G3bp2 were upregulated, while miR-185-5p was downregulated in DN mice and HG-treated podocytes.
- HG treatment impaired podocyte proliferation and induced apoptosis and inflammation; circ_Supt3 knockdown reversed these effects.
- Circ_Supt3 directly binds to miR-185-5p, and miR-185-5p targets G3bp2, forming a regulatory axis.
Conclusions:
- Circ_Supt3 exacerbates high glucose-induced podocyte injury by sponging miR-185-5p and upregulating G3bp2.
- Knockdown of circ_Supt3 demonstrates a protective effect against DN-related podocyte damage via the miR-185-5p/G3bp2 pathway.
- The circ_Supt3/miR-185-5p/G3bp2 axis represents a potential therapeutic target for diabetic nephropathy.
