Figure 6.

Colchicine inhibits the adhesion of HUVECs with monocytes by regulating the TLR4 pathway. (a, b) HUVECs in 24-well plates were treated with LPS (1 mg/mL), colchicine (100 ng/mL), or LPS plus colchicine for 24 h (a), or ox-LDL (100 μg/mL), colchicine (100 μg/mL), or ox-LDL plus colchicine overnight (b). After treatment, the CFSE-labeled THP-1 monocytes (1 × 10⁵ cells/well) were added to HUVECs and incubated for 1 h at 37°C. After washing away the unadhered THP-1 cells with PBS, labeled cells were photographed by a fluorescence microscope. The number of THP-1 cells in each group was counted and normalized to the number in the control group; (c–f) HUVECs (c) or RAW264.7 cells (d–f) were treated with ticagrelor (10 mM), colchicine (100 ng/mL), or ticagrelor plus colchicine in the presence of LPS (500 ng/mL) for 24 h. Expression of ICAM-1, VCAM-1 (c), and TNF-α (d) mRNA was determined by qRT-PCR. Expression of IL-1β, TLR4, and GAPDH was determined by Western blot (e) with quantitation of band density (f). ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001 vs. control; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. LPS or ox-LDL alone (n = 3).