Colchicine Alleviates Cholesterol Crystal-Induced Endothelial Cell Pyroptosis through Activating AMPK/SIRT1 Pathway

Figure 4

Colchicine plays an antipyroptosis role by inhibiting intracellular oxidative stress. HUVECs were subjected to H₂O₂ (300 μM), cholesterol crystal (0.5 mg/ml) and colchicine (10 nM), or N-acetyl-cysteine (NAC, 5 mM) for 10 h or 24 h. (a) ROS level was detected using a DCFH-DA probe, and (b) mitochondrial membrane potential was tested by JC-1 (). (c, d) The indicators of oxidative stress GSSG and MDA were tested by assay kits. (e) Western blotting was dedicated to examine the protein expression levels of NLRP3, ASC-1, procaspase-1, and caspase-1. (f–h) Quantitative analysis of pyroptosis-associated protein expression. (i, j) Real-time PCR analysis of the mRNA levels of GSDMD and proinflammatory cytokines (IL-18 and IL-1β). Data was expressed as the of three separate experiments. , , and vs. the control group. ^##, ^###, and ^#### vs. the cholesterol crystal group. ^&, ^&&, ^&&&, and ^&&&& vs. the H₂O₂ group.