Efficacy and molecular mechanism of paclitaxel-bivalirudin interventional coating complex against vascular restenosis based on NF-κB and NRF2/ARE pathways

AIM To investigate the effect of paclitaxel-bivaludin interventional coating combination (LFN) on vascular restenosis and its molecular mechanism.

METHODS In vivo animal experiment grouping: WT sham surgery group, WT vascular strain group, NRF2^-/- vascular strain group, NF-κB^-/- vascular strain group, WT vascular strain+LFN intervention group, NRF2^-/- vascular strain+LFN intervention group, and NF-κB^-/- vascular strain+LFN intervention group; In vitro cell experiment grouping: First batch of groups: Control group, LPS modeling group, LPS+LFN group, LPS+LFN+NF-κB knockdown group, LPS+LFN+IκB knockdown group, LPS+LFN+NF-κB over-expression group, and LPS+LFN+IκB over-expression group. Second batch of groups: Control group, LPS modeling group, LPS+LFN group, LPS+LFN+NRF2 knockdown group, LPS+LFN+Keap1 knockdown group, LPS+LFN+NRF2 over-expression group, and LPS+LFN+Keap1 over-expression group. The pathological changes of vascular tissue were detected using HE staining, the proliferation activity of vascular tissue was detected using immunofluorescence staining, the cell proliferation rate was detected using the CCK-8 method, the cell migration and invasion ability was detected using the Transwell method, and the gene and protein expressions of important targets and ligands of the NF-κB and NRF2/ARE pathways were determined in vascular tissues and cells using Q-PCR and Western blot.

RESULTS In vascular strain model rats, LFN significantly inhibited intimal hyperplasia, which effectively prevented lumen restenosis and combated thrombosis simultaneously. LFN up-regulated the expressions of IκB, NRF2, NQO-1 and HO-1 genes and proteins compared with those in WT vascular strain group (P<0.05, P<0.01), but down-regulated the expressions of Keap-1 genes and proteins (P<0.05, P<0.01), which were reversed in NRF2^-/- mutant rats. The antagonistic effect of LFN on the HCASMC model’s capacity for migration and invasion was affected by NF-κB, NRF2, with its ligands being knocked down or over-expressed. This also, to variable degrees, impaired LFN’s capacity to regulate the production of crucial proteins and genes in the NF-κB and NRF2/ARE pathways in cells.

CONCLUSION LFN is efficacious in vivo against vascular restenosis and part of the effect is achieved by the unified regulation of the expressions of NF-κB and NRF2/ARE nuclear transcription factors and their essential ligands.