1,8-cineole improved lipopolysaccharide-induced inflammatory injury in human umbilical vein endothe-lial cells via TLR4/NF-κB signaling pathway
OBJECTIVE To explore the protective mechanism of 1,8-cineole antagonizing inflammatory injury in human umbilical vein endothelial cells(HUVECs).METHODS CCK-8 was utilized for detecting different concentrations of 1,8-cin-eole(0.08,0.12,0.31,0.63,1.25,2.50,5.00,10.00 μg·L-1)for screening the optimal concentration.HUVECs were exposed to lipopolysaccharide(LPS).HUVEC cells were assigned into six groups of control,model,low/medium/high-dose 8-cineole and medium-dose8-cineole plus KLA.Cellular viability,apoptosis and migration were detected by CCK-8,flow cytom-etry and transwell.The contents of malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by kits.The mRNA contents of anti-intercellular adhesion molecule-1(ICAM-1)and anti-vascular cell adhesion molecule-1(VCAM-1)were detected by polymerase chain reaction(PCR).The expression levels of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),Toll-like receptor 4(TLR4),cytoplasmic nuclear factor-kappa B p65(NF-κB p65),nuclear NF-κB p65 and cleaved cas-pase-3 protein were detected by Westem blot.RESULTS As compared with control group,inhibition rate,apoptotic rate,MDA content,IL-6/TNF-α protein expression,ICAM-1/CAM-1 mRNA expression,NF-κB p65 nuclear translocation,TLR4,nuclear NF-κB p65 and cleaved caspase-3 protein expression rose obviously in model group(P＜0.05)while migration number,SOD and cytoplasmic NF-κB p65 protein expression dropped markedly(P＜0.05).As compared with model group,cel-lular inhibition rate,apoptotic rate,MDA content,IL-6/TNF-α protein expression,NF-κB p65 nuclear translocation,TLR4,nuclear NF-κB p65 and cleaved Caspase-3 protein expression decreased significantly(P＜0.05)while migration number,SOD and cytoplasmic NF-κB p65 protein expression spiked significantly in low/middle/high-dose group(P＜0.05).The mRNA expression of ICAM-1/VCAM-1 declined markedly in middle/high-dose group(P＜0.05).All parameters were worse in medium-dose+KLA group than those in medium-dose group.CONCLUSION Inflammatory injury of vascular endothelial cells induced by LPS may be correlated with nuclear trarislocation of NF-κB p65 in TLR4/NF-κB pathway.And 1,8-cineole may treat vascular endothelial cell injury through suppressing NF-κB p65 nuclear translocation,blunting inflammatory response,lowering oxidative stress and reducing vascular endothelial cell adhesion.
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