Fig. 3

miR-711 negatively regulates TLR4 and reduces high glucose-provoked inflammatory responses of H9c2 cells. A qRT-PCR was used to detect the expression of TLR4 mRNA in H9c2 cells treated with mannitol or high glucose. Western blotting was used to detect the expression of TLR4 protein B and p-NF-κB p65 protein C in H9c2 cells treated with mannitol or high glucose. D TargetScan database predicted the binding sites between miR-711 and TLR4 mRNA and mutations were designed. E Dual-luciferase reporter assay was performed to verify the binding between miR-711 and TLR4 mRNA. Next, high glucose-treated H9c2 cells were transfected with mimic-NC, miR-711 mimic, inhibitor-NC or miR-711 inhibitor. F qRT-PCR was used to detect the expression of miR-711 in the transfected cells. Western blotting was used to detect the expression of TLR4 protein G and p-NF-κB p65 protein H in the transfected cells. I ELISA was performed to detect the concentrations of ICAM-1, MCP-1, and IL-1β in cell culture supernatant. The data were all measurement data and expressed as mean ± standard deviation. One-way analysis of variance was used for comparisons between multiple groups and Tukey’s test for post-hoc multiple comparisons. ^* indicates P < 0.05. The experiments were repeated thrice. OS osmotic, HG high glucose, NC negative control, TLR4 toll-like receptor 4, qRT-PCR quantitative real-time polymerase chain reaction, ELISA enzyme-linked immunosorbent assay, ICAM intercellular adhesion molecule, MCP monocyte chemotactic protein, IL interleukin