Metformin regulates inflammation and fibrosis in diabetic kidney disease through TNC/TLR4/NF-κB/miR-155-5p inflammatory loop

Figure 3 Changes in the levels of the tenascin-C and toll-like receptor-4 proteins and nuclear factor-κB p65 protein (Ser536) phosphorylation in cells stimulated with high glucose. A and B: Protein bands and protein expression of tenascin-C (TNC). Rat mesangial cells (RMCs) were cultured under normal (NG, 5.5 mmol/L glucose) and hypertonic (HM, 5.5 mmol/L glucose + 24.5 mmol/L mannitol) conditions for 72 h, or high-glucose (HG, 30 mmol/L glucose) conditions for 24, 48, and 72 h. ^aP < 0.05 compared with the 72 h control group; C-G: Protein bands and protein expression of TNC, Toll-like receptor-4, and phosphorylated nuclear factor-κB p65 protein (Ser536). RMCs were cultured under normal (NG, 5.5 mmol/L glucose), hypertonic (HM, 5.5 mmol/L glucose + 24.5 mmol/L mannitol), and high-glucose (HG, 30 mmol/L glucose) conditions for 24 h. ^aP < 0.05 compared with the N control group. Protein levels were detected using Western blot. The results are presented as the mean ± SD of six independent experiments after normalization to GAPDH levels. TNC: Tenascin-C; TLR4: Toll-like receptor-4; p-NF-κB p65: Phosphorylated nuclear factor-κB p65 protein (Ser536); N: Normal glucose; M: Hypertonic; H: High-glucose.