Department of Endocrinology, The People's Hospital of Jiaozuo City, Jiaozuo 454000, China.
Department of General Surgery, Jiaozuo Hospital of Traditional Chinese Medicine, Jiaozuo 454000, China.
Diabetes Res Clin Pract. 2021 Jun;176:108837. doi: 10.1016/j.diabres.2021.108837. Epub 2021 Apr 24.
CircRNAs are reported to be aberrantly expressed and perform biological functions in diverse processes. This study aimed to investigate the potential involvement of hsa_circ_0054633 in high glucose (HG)‑induced diabetic model and its potential mechanism.
The expression of hsa_circ_0054633, miR-409-3p and caspase-8 was detected by real-time PCR and western blotting. Cell viability, apoptosis and the protein levels of apoptosis-related factors were revealed by CCK-8 colorimetry, flow cytometry and western blotting, respectively. Insulin secretion was determined by enzyme-linked immunosorbent assay (ELISA) and the measurement of insulin-related transcription factors. The target association between miR-409-3p and hsa_circ_0054633 or caspase-8 was confirmed by dual-luciferase reporter assays and biotin-based pulldown assay.
Hsa_circ_0054633 was highly expressed and the expression of miR-409-3p was downregulated in serum of DM patients and HG-treated human pancreatic β cell line NES2Y. Further investigation indicated that hsa_circ_0054633 suppression promoted cell proliferation, inhibited apoptosis and facilitated insulin secretion in HG-treated NES2Y cells. Mechanical analysis revealed that hsa_circ_0054633 regulated caspase-8 expression via sponging miR-409-3p. Rescue experiments demonstrated that miR-409-3p knockdown or caspase-8 overexpression reversed the effects of hsa_circ_0054633 in HG-stimulated NES2Y cells.
Inhibition of hsa_circ_0054633 protected against HG-induced NES2Y cell apoptosis and impairment of insulin secretion by regulating miR-409-3p/caspase-8 axis.
CircRNAs 被报道在多种过程中异常表达并发挥生物学功能。本研究旨在探讨 hsa_circ_0054633 在高糖（HG）诱导的糖尿病模型中的潜在作用及其潜在机制。
通过实时 PCR 和 Western blot 检测 hsa_circ_0054633、miR-409-3p 和 caspase-8 的表达。通过 CCK-8 比色法、流式细胞术和 Western blot 分别揭示细胞活力、凋亡和凋亡相关因子的蛋白水平。通过酶联免疫吸附测定（ELISA）和胰岛素相关转录因子的测定来确定胰岛素分泌。通过双荧光素酶报告基因检测和生物素 pulldown assay 证实 miR-409-3p 与 hsa_circ_0054633 或 caspase-8 的靶标关联。
在 DM 患者的血清和 HG 处理的人胰腺β细胞系 NES2Y 中，hsa_circ_0054633 表达上调，miR-409-3p 表达下调。进一步的研究表明，hsa_circ_0054633 抑制促进了 HG 处理的 NES2Y 细胞的增殖，抑制了凋亡，并促进了胰岛素的分泌。机制分析表明，hsa_circ_0054633 通过海绵吸附 miR-409-3p 调节 caspase-8 的表达。挽救实验表明，miR-409-3p 敲低或 caspase-8 过表达逆转了 hsa_circ_0054633 在 HG 刺激的 NES2Y 细胞中的作用。
抑制 hsa_circ_0054633 通过调节 miR-409-3p/caspase-8 轴，防止 HG 诱导的 NES2Y 细胞凋亡和胰岛素分泌受损。
