Abstract
Diabetic nephropathy (DN) threatens the survival quality of patients, with complex pathogenesis. Circular RNA (circRNA) dysregulation occurs in DN development. This work aimed to investigate the role of circ-Luc7l in DN cell models and related molecular mechanisms. The expression of circ-Luc7l, microRNA (miR)-205-5p, and transforming growth factor-beta receptor 1 (Tgfbr1) was examined by real-time quantitative PCR (RT-qPCR). Cell viability and proliferation were detected by Cell Counting Kit-8 (CCK-8) assay and EdU assay. The expression of extracellular matrix (ECM)-related markers and Tgrbr1 protein was measured by Western blot. The binding between miR-205-5p and circ-Luc7l or Tgfbr1 was validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, or RNA pull-down assay. Experimental animal models were established to elucidate the function of circ-Luc7l in vivo. Circ-Luc7l expression was notably enhanced in high glucose (HG)-treated mesangial cells. Knockdown of circ-Luc7l attenuated HG-induced cell proliferation, inflammation, and ECM accumulation in vitro and relieved inflammation and ECM accumulation of kidneys of diabetic mice in vivo. Circ-Luc7l targeted miR-205-5p, and miR-205-5p inhibition rescued the depletion effects of circ-Luc7l knockdown on cell proliferation, inflammation, and ECM accumulation. MiR-205-5p bound to Tgfbr1 whose expression was negatively regulated by circ-Luc7l. Tgfbr1 overexpression also rescued the depletion effects of circ-Luc7l knockdown on cell proliferation, inflammation, and ECM accumulation. In HG conditions, increased circ-Luc7l upregulated Tgfbr1 expression via targeting miR-205-5p to induce DN progression.
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ZF designed and performed the research; DW, FS, JC, DY, SL, and JT analyzed the data; ZF wrote the manuscript. All authors read and approved the final manuscript.
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Fig. S1 Regulation of the circ-Luc7l/miR-205-5p/Tgfbr1 axis in proliferation, inflammation, and ECM accumulation of MC53 cells. A RT-qPCR for circ-Luc7l/miR-205-5p/Tgfbr1 mRNA expression in MC53 cells treated as indicated and then in HG-treated SV40-MES13 cells transfected with si-circ-Luc7l, si-circ-Luc7l+anti-miR-205-5p, or si-circ-Luc7l+Tgfbr1. B Cell viability was ascertained by CCK-8 assay. C Cell proliferation was determined by EdU assay. D, E The release of IL-6 and TNF-α was examined using commercial ELISA kits. F The protein levels of FN, Col I, and α-SMA were determined by Western blot. *P < 0.05, **P < 0.01, and ***P < 0.001.
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Fang, Z., Wang, D., Sun, F. et al. Circ-Luc7l Absence Attenuates Diabetic Nephropathy Progression by Reducing Mesangial Cell Excessive Proliferation, Inflammation, and Extracellular Matrix Accumulation via Mediating the miR-205-5p/Tgfbr1 Pathway. Biochem Genet (2024). https://doi.org/10.1007/s10528-024-10694-9
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DOI: https://doi.org/10.1007/s10528-024-10694-9