Department of Intensive Care Unit, The First Hospital of Jilin University, Changchun, Jilin, 130021, China.
Department of Neurosurgery, China-Japan Union Hospital of Jilin University, Jilin University, Changchun, Jilin, 130033, China.
Biochem Biophys Res Commun. 2020 Nov 5;532(2):308-314. doi: 10.1016/j.bbrc.2020.08.073. Epub 2020 Aug 29.
This study aimed to investigate the involvement of lncRNA CTBP1-AS2 in the progression of diabetic nephropathy (DN) by affecting high glucose (HG)-induced human glomerular mesangial cells (HGMCs).
HGMCs were selected for the establishment of cell injury induced by HG. The expression of CTBP1-AS2, miR-155-5p and FOXO1 was detected by real-time PCR and western blotting. The target association between miR-155-5p and CTBP1-AS2 or FOXO1 was confirmed by dual-luciferase reporter assays. Cell proliferation and oxidative stress were revealed by CCK-8 colorimetry, and the measurement of reactive oxygen species (ROS) and the activities of antioxidant enzymes. Extracellular matrix (ECM) protein accumulation and the production of inflammatory cytokines were investigated by western blotting and ELISA.
The expression of CTBP1-AS2 was downregulated, and miR-155-5p was highly expressed in peripheral blood of DN patients and HG-treated HGMCs. Further investigation revealed that CTBP1-AS2 overexpression inhibited proliferation, oxidative stress, ECM accumulation and inflammatory response in HG-induced HGMCs. Mechanical analysis revealed that CTBP1-AS2 regulated FOXO1 expression via sponging miR-155-5p. Rescue experiments demonstrated that miR-155-5p overexpression or FOXO1 inhibition reversed the effects of CTBP1-AS2 in HG-stimulated HGMCs.
Taken together, this study revealed CTBP1-AS2 attenuated HG-induced HGMC proliferation, oxidative stress, ECM accumulation, and inflammation through miR-155-5p/FOXO1 signaling.
本研究旨在通过影响高糖（HG）诱导的人肾小球系膜细胞（HGMCs），探讨 lncRNA CTBP1-AS2 在糖尿病肾病（DN）进展中的作用。
选择 HGMCs 建立 HG 诱导的细胞损伤模型。采用实时 PCR 和 Western blot 检测 CTBP1-AS2、miR-155-5p 和 FOXO1 的表达。采用双荧光素酶报告基因实验验证 miR-155-5p 与 CTBP1-AS2 或 FOXO1 的靶基因关系。采用 CCK-8 比色法检测细胞增殖和氧化应激，检测活性氧（ROS）和抗氧化酶的活性。采用 Western blot 和 ELISA 检测细胞外基质（ECM）蛋白积累和炎症细胞因子的产生。
DN 患者外周血和 HG 处理的 HGMCs 中 CTBP1-AS2 表达下调，miR-155-5p 表达上调。进一步研究表明，CTBP1-AS2 过表达抑制 HG 诱导的 HGMCs 增殖、氧化应激、ECM 积累和炎症反应。机制分析表明，CTBP1-AS2 通过海绵吸附 miR-155-5p 调节 FOXO1 表达。挽救实验表明，miR-155-5p 过表达或 FOXO1 抑制逆转了 CTBP1-AS2 在 HG 刺激的 HGMCs 中的作用。
综上所述，本研究表明 CTBP1-AS2 通过 miR-155-5p/FOXO1 信号通路减轻 HG 诱导的 HGMCs 增殖、氧化应激、ECM 积累和炎症反应。
