Department of Nephrology, Shenzhen Nanshan People's Hospital and The 6Th Affiliated Hospital of Shenzhen University Medical School, Shenzhen, China.
Appl Biochem Biotechnol. 2023 Dec;195(12):7255-7276. doi: 10.1007/s12010-023-04431-y. Epub 2023 Mar 29.
Emerging evidences suggested that circular RNAs (circRNAs) are involved in diabetic nephropathy (DN). Accumulating evidence had suggested that the degree of podocyte is a major prognostic determinant of DN progression. However, the function and in-depth mechanisms of hsa_circ_0001162 in podocyte injury of DN remain unclear. Hsa_circ_0001162 expression was detected by real-time quantitative PCR (RT-qPCR) in peripheral blood of DN patients and high glucose-induced podocytes injury model. The cell counting kit 8, 5-ethynyl-2'-deoxyuridine, flow cytometry with Annexin V-FITC/PI staining, caspase-3 activity assay Kit, enzyme linked immunosorbent assay (ELISA), RT-qPCR and western blotting were used to evaluate the effect of hsa_circ_0001162 / miR-149-5p / MMP9 axis on high glucose-induced podocyte injury. Mechanistically, dual luciferase reporter was used to confirm the relationship of miR-149-5p and hsa_circ_0001162 or MMP9. Furthermore, RNA-pull down and immunoprecipitation assay were implemented to verify the potential regulatory effects of EIF4A3 on biogenesis of hsa_circ_0001162. Our results showed that hsa_circ_0001162 was highly expressed in peripheral blood of DN patients and high glucose-induced podocytes injury model, and the knockdown of hsa_circ_0001162 increased the proliferation, inhibited the apoptosis, and suppressed inflammatory response in high glucose-induced podocytes injury. Mechanism studies demonstrated that EIF4A3 bound with flanking sequences of hsa_circ_0001162 to promote hsa_circ_0001162 expression, upregulated hsa_circ_0001162 increased the MMP9 expression via sponging miR-149-5p, thus aggravating the high glucose-induced podocytes injury. Overall, our data demonstrated that knockdown of hsa_circ_0001162 inhibited high glucose-induced podocytes injury by regulating miR-149-5p/MMP9 axis, and intervention of hsa_circ_0001162/miR-149-5p/MMP9 axis may be a potentially promising therapeutic strategy for podocyte injury in DN patients.
越来越多的证据表明，环状 RNA（circRNA）参与了糖尿病肾病（DN）的发生。越来越多的证据表明，足细胞的程度是 DN 进展的主要预后决定因素。然而，hsa_circ_0001162 在 DN 足细胞损伤中的功能和深入机制仍不清楚。通过实时定量 PCR（RT-qPCR）检测 DN 患者外周血和高糖诱导的足细胞损伤模型中 hsa_circ_0001162 的表达。使用细胞计数试剂盒 8、5-乙炔基-2'-脱氧尿苷、流式细胞术结合 Annexin V-FITC/PI 染色、caspase-3 活性测定试剂盒、酶联免疫吸附试验（ELISA）、RT-qPCR 和 Western blot 评估 hsa_circ_0001162/miR-149-5p/MMP9 轴对高糖诱导的足细胞损伤的影响。从机制上讲，双荧光素酶报告基因用于证实 miR-149-5p 与 hsa_circ_0001162 或 MMP9 的关系。此外，实施 RNA 下拉和免疫沉淀测定以验证 EIF4A3 对 hsa_circ_0001162 生物发生的潜在调节作用。我们的结果表明，hsa_circ_0001162 在 DN 患者外周血和高糖诱导的足细胞损伤模型中高表达，hsa_circ_0001162 的敲低增加了增殖，抑制了凋亡，并抑制了高糖诱导的足细胞损伤中的炎症反应。机制研究表明，EIF4A3 与 hsa_circ_0001162 的侧翼序列结合，以促进 hsa_circ_0001162 的表达，上调 hsa_circ_0001162 通过海绵 miR-149-5p 增加 MMP9 的表达，从而加重高糖诱导的足细胞损伤。总的来说，我们的数据表明，通过调节 miR-149-5p/MMP9 轴，敲低 hsa_circ_0001162 抑制了高糖诱导的足细胞损伤，hsa_circ_0001162/miR-149-5p/MMP9 轴的干预可能是 DN 患者足细胞损伤的一种有前途的潜在治疗策略。
