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Published on: August 23, 2024
Circ_0000524/miR-500a-5p/CXCL16 axis promotes podocyte apoptosis in membranous nephropathy
Zhiqiang Sun1, Qingqing Xu1, Yali Ma1
1Department of Nephrology, Huaihe Hospital of Henan University, Kaifeng, China.
Background:
Podocytes apoptosis is a hallmark of membranous nephropathy (MN). Circ_0000524 has been reported to be associated with patients with MN, whereas the effect of circ_0000524 on podocytes apoptosis and the underlying mechanisms in MN have not been elaborated.
Methods:
Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were performed to detect the expressions of circ_0000524, microRNA-500a-5p (miR-500a-5p), and C-X-C chemokine ligand 16 (CXCL16) in MN tissues and podocytes. Podocyte injury was induced by angiotensin II (AngII). Cell apoptosis was detected by flow cytometry. Caspase-3 or caspase-9 activity was evaluated using a caspase-3 or caspase-9 activity assay kit, respectively. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) and pull-down assay were used to address the relationship among circ_0000524,miR-500a-5p and CXCL16.
Results:
Upregulation of circ_0000524 and CXCL16 and low expression of miR-500a-5p were observed in MN tissues. AngII treatment induced the overexpression of circ_0000524 and CXCL16, a decrease of miR-500a-5p, and induced cell apoptosis in podocytes. Circ_0000524 negatively modulated the expression of miR-500a-5p. Circ_0000524 depletion inhibited podocyte apoptosis, which was rescued by loss of miR-500a-5p. miR-500a-5p contained the binding sites with CXCL16. Circ_0000524 knockdown hampered CXCL16 expression by upregulating miR-500a-5p expression. Additionally, miR-500a-5p upregulation suppressed AngII-induced podocyte apoptosis, which was rescued by enhanced expression of CXCL16.
Conclusion:
Circ_0000524/miR-500a-5p/CXCL16 pathway regulated podocyte apoptosis in MN.
Insights
Circ_0000524 promotes podocyte apoptosis in membranous nephropathy (MN) by regulating the miR-500a-5p/CXCL16 pathway. Inhibiting circ_0000524 may offer a therapeutic strategy for MN.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Podocyte apoptosis is a key feature of membranous nephropathy (MN).
- Circ_0000524 is linked to MN, but its role in podocyte apoptosis and underlying mechanisms remain unclear.
Purpose of the Study:
- To investigate the role of circ_0000524 in podocyte apoptosis in MN.
- To elucidate the molecular mechanisms involving circ_0000524, miR-500a-5p, and CXCL16 in MN.
Main Methods:
- Quantitative real-time PCR and Western blot to assess gene and protein expression.
- Angiotensin II (AngII) to induce podocyte injury.
- Flow cytometry for apoptosis analysis.
- Dual-luciferase reporter, RNA immunoprecipitation, and pull-down assays to determine molecular interactions.
Main Results:
- Circ_0000524 and CXCL16 were upregulated, while miR-500a-5p was downregulated in MN tissues and AngII-treated podocytes.
- Circ_0000524 depletion reduced podocyte apoptosis, an effect reversed by miR-500a-5p inhibition.
- MiR-500a-5p directly targeted CXCL16, and circ_0000524 knockdown increased miR-500a-5p, thereby decreasing CXCL16.
- MiR-500a-5p upregulation suppressed apoptosis, which was reversed by CXCL16 overexpression.
Conclusions:
- The circ_0000524/miR-500a-5p/CXCL16 pathway plays a crucial role in regulating podocyte apoptosis in MN.
- Targeting this pathway may represent a novel therapeutic approach for MN.
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