Effect of Formononetin on Human Coronary Endothelial Cell Against ox-LDL-induced Injury by Regulating TLR4/NF-κB Pathway
YUAN Xiaocui
LIAO Jinyu
FU Li
LUO Yong
CHEN Dong
Abstract：Objective:To investigate the effect of formononetin(Form)in regulating toll-like receptor 4(TLR4)/nuclear factor κB(NF-κB)pathway on human coronary endothelial cell(HCAECs)against oxidative low-density lipoprotein(ox-LDL)-induced injury.Methods:The HCAECs treated with 24 μg/mL ox-LDL for 40 h induced injury and inflammation,and the non-ox-LDL treated HCAECs served as the control group.HCAECs treated with ox-LDL were divided into HCAECs group,L-form group(16 μmol/L Form),M-Form group(32 μmol/L Form),H-Form group(64 μmol/L Form)and lipopolysaccharide(LPS)group(5 μg/mL)TLR4/NF-κB p65 pathway activator LPS)and H-Form+LPS groups(64 μmol/L Form+5 μg/mL LPS).Cell proliferation activity was detected by cell counting(CCK-8).Flow cytometry was used to detect apoptosis.Cell migration and invasion were measured by Transwell method.Matrix glue was used to evaluate tube formation ability.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Western Blot was used to detect TLR4/NF-κB pathway protein expression.Results:Compared with the control group,the apoptosis rate,the levels of IL-1β,IL-6,TNF-α,the expression of TLR4,P-NF-κB p65/NF-κB p65 protein in HCAECs group significantly increased(P＜0.05),and the OD value of cell proliferation activity significantly decreased(P＜0.05),the number of migrating cells,the number of invading cells,the number of capillary tube nodes significantly decreased(P＜0.05),and the total tube length significantly reduced(P＜0.05).Compared with HCAECs group,the apoptosis rate of HCAECs cells,the levels of IL-1β,IL-6,and TNF-α,and the expression of TLR4 and P-NF-κB p65/NF-κB p65 protein in L-Form,M-Form,and H-Form groups significantly decreased(P＜0.05).OD value of cell proliferation activity,number of migrating cells,number of invading cells,number of capillary tube nodes,and total tube length significantly increased(P＜0.05),but the trend was opposite in LPS group.LPS eliminated the beneficial effect of high concentration Form on HCAECs cells.Conclusion:Form may alleviate ox-LDL-induced HCAECs damage by inhibiting the TLR4/NF-κB signaling pathway.
Keywords：formononetinhuman coronary artery endothelial cellsoxidative low-density lipoproteinToll-like receptor 4/nuclear factor κB pathwayexperimental study
Publication Date：2025-04-10
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 1015-1021 )