Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, India.
Department of General Medicine, Sri Devaraj Urs Medical College, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, India.
Braz J Med Biol Res. 2022 Jul 25;55:e11984. doi: 10.1590/1414-431X2022e11984. eCollection 2022.
The aim of this study was to evaluate the impact of N6-carboxymethyllysine (CML) on NF-κB gene expression and tumor necrosis factor (TNF) production in diabetic nephropathy. This was an observational study comprised of three groups: diabetic nephropathy (n=30), type II diabetes mellitus (n=28), and healthy volunteers (n=30). Blood samples collected from the study participants were cultured for 24 h in the presence of CML or an appropriate control. After incubation, the cultures were centrifuged to separate the cells from the conditioned media. cDNA was prepared from the cell pellet and used to quantify NF-κB gene expression by quantitative real-time polymerase chain reaction (PCR). The conditioned media were used to measure TNF production by enzyme-linked immunosorbent assay (ELISA). The CML-induced fold change in NF-κB gene expression was significantly different among the study groups (P=5.4×10-5). Also, the CML-induced fold change in TNF levels was significantly different among the three groups (P=4.3×10-8). These results imply that patients with diabetic nephropathy and type II diabetes mellitus showed an elevated response to CML.
本研究旨在评估 N6-羧甲基赖氨酸 (CML) 对糖尿病肾病中 NF-κB 基因表达和肿瘤坏死因子 (TNF) 产生的影响。这是一项观察性研究，包括三组：糖尿病肾病 (n=30)、2 型糖尿病 (n=28) 和健康志愿者 (n=30)。从研究参与者采集的血液样本在存在 CML 或适当对照的情况下培养 24 小时。孵育后，将培养物离心以将细胞与条件培养基分离。从细胞沉淀中制备 cDNA，并通过实时定量聚合酶链反应 (PCR) 定量 NF-κB 基因表达。使用酶联免疫吸附测定 (ELISA) 测量条件培养基中 TNF 的产生。CML 诱导的 NF-κB 基因表达倍数变化在研究组之间有显著差异 (P=5.4×10-5)。此外，CML 诱导的 TNF 水平的倍数变化在三组之间有显著差异 (P=4.3×10-8)。这些结果表明，糖尿病肾病和 2 型糖尿病患者对 CML 的反应升高。
