Cafestol Inhibits High-Glucose-Induced Cardiac Fibrosis in Cardiac Fibroblasts and Type 1-Like Diabetic Rats

Figure 3

Molecular docking of cafestol with Keap1. (a) The structure of the receptor protein Keap1 (PDB: 1U6D). (b) Representative binding modes of the cafestol molecule in the active pocket of Keap1. (c) The pocket reveals that cafestol inserted into the cavity of Keap1 and the hydroxyl group of cafestol formed two hydrogen bonds with the backbone Val465 and Ile559. (d) Hydrophobic interactions and hydrogen bonding between cafestol and Keap1. LigPlot⁺ was used for visualization of protein-ligand interactions. Hydrogen bonds are illustrated as green dashed lines, and hydrophobic interactions are illustrated as red arcs. (e) Cafestol binds to KEAP1 to release Nrf2. Cardiac fibroblasts pretreated with MG132 (10 μM) for 2 h then lysed with lysis buffer. The cell lysates were incubated at 30°C for 2 h in the presence or absence of cafestol (30 μM) followed by using immunoprecipitation kits (Thermo Fisher Scientific) with anti-Nrf2 antibody. The Nrf2 and KEAP1 proteins in the immunocomplex were monitored by western blot analysis. Representative western blot of Nrf2 and Keap-1 protein expression (n = 3). IP: immunoprecipitation. IB: immunoblotting.

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