Department of Nephrology, Shanghai TCM-Integrated Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Department of Endocrinology, Shanghai Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Shock. 2024 Aug 1;62(2):227-234. doi: 10.1097/SHK.0000000000002389. Epub 2024 May 23.
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.
糖尿病肾病（DN）是糖尿病的一种并发症，是糖尿病患者死亡的主要原因。环状 RNA（circRNA）是人类疾病研究的热门话题。然而，circ_Supt3 在 DN 中的作用尚不清楚。方法：用高葡萄糖（HG）处理小鼠足细胞（MPC5）细胞模拟 DN 细胞损伤。实时定量聚合酶链反应检测 circ_Supt3、微小 RNA-185-5p（miR-185-5p）和 G 蛋白激活蛋白结合蛋白 2（G3bp2）的表达。5-乙炔基-2'-脱氧尿苷（EdU）和 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）测定法检测细胞增殖，流式细胞术检测细胞凋亡。Western blot 法检测相对蛋白水平。酶联免疫吸附试验检测炎症细胞因子。双荧光素酶报告和 RNA 下拉实验证实 miR-185-5p 与 circ_Supt3 或 G3bp2 的相互作用。结果：DN 小鼠中 circ_Supt3 和 G3bp2 表达升高，miR-185-5p 表达降低。HG 处理抑制细胞增殖，加速细胞凋亡和炎症反应，而 circ_Supt3 的敲低逆转了这些作用。生物信息学预测 circ_Supt3 含有 miR-185-5p 的结合位点，G3bp2 是 miR-185-5p 的直接靶点。circ_Supt3 通过 miR-185-5p 调节 G3bp2 的表达。此外，circ_Supt3/miR-185-5p/G3bp2 轴调节 HG 诱导的 MPC5 细胞的细胞行为。结论：我们的研究结果表明，circ_Supt3 的敲低通过 miR-185-5p/G3bp2 轴保护 HG 诱导的 MPC5 细胞免受细胞损伤。
