Effect of matrine on lipopolysaccharide-induced inflammation and oxidative stress in human umbilical vein endothelial cells

Abstract

Objective To investigate the effects of matrine (MAT) on lipopolysaccharide (LPS) -induced inflammation and oxidative stress in human umbilical vein endothelial cells (HUVECs) . Methods HUVECs were randomly divided into 5 groups: control group, LPS group, LPS+MAT low, medium and high dose treatment groups. In the LPS group, the model of inflammation and oxidative stress was established by stimulation with 100 ng/ml LPS for 12 h. In the LPS+MAT low, medium and high-dose treatment groups, HUVECs were pretreated with different concentrations of MAT (0.5, 1.0, and 1.5 mmol/L) for 24 h, and then incubated with addition of LPS (100 ng/ml) for 12 h in the incubator. The viability of HUVECs was detected by CCK-8 assay. The mRNA expression levels of IL-1β, IL-6, MCP-1, TNF-α, ICAM-1 and VCAM-1 were detected by real-time fluorescent quantitative PCR. The adhesion of THP-1 cells to HUVECs was detected by microplate reader. Western blotting was used to detect the protein expression of ICAM-1, VCAM-1 and NF-κB P65. The DCFH-DA probe combined with the microplate reader was used to detect the reactive oxygen species (ROS) content in the cells. Intracellular dialdehyde (MDA) content, activity of glutathione peroxidase (GSH-Px) and catalase (CAT) were measured with specific tesk kits. Results MAT was found to exhibit cytotoxicity at levels ≥ 2.0 mmol/L, and reduce HUVECs viability in a dose-dependent manner (P<0.05) . Compared with the control group, the LPS group showed elevated mRNA levels of IL-1β, IL-6, MCP-1, TNF-α, ICAM-1 and VCAM-1, higher rate of THP-1 cell adhesion to HUVECs, increased protein expression of ICAM-1, VCAM-1 and P65 protein expression, higher contents of ROS and MDA, and lowered activities of GSH-Px and CAT. MAT pretreatment was shown to inhibit LPS-induced inflammatory response in endothelial cells, reduce THP-1 cell adhesion to HUVECs, and improve oxidative stress. These effects were more prominent with higher doses (P<0.05) . Conclusion MAT exhibits protective effect against LPS-induced HUVECs injury, which may be related to reduction of inflammatory cytokines expression and inhibition of oxidative stress. Key words: Matrine; Lipopolysaccharide; Human umbilical vein endothelial cells; Inflammation; Oxidative stress