Effects of Lycium barbarum polysaccharide on lipopolysaccharide-induced endothelial cell dysfunction and AKT/eNOS pathway
OBJECTIVE To explore the effect and mechanism of Lycium barbarum polysaccharide(LBP)on endothelial cell dysfunction induced by sepsis.METHODS Human umbilical vein endothelial cells(HUVECs)were selected and endothe-lial cell dysfunction model was induced by lipopolysaccharide(LPS).Cellular morphology was observed under microscope,cellu-lar viability was detected by CCK8 and the levels of C-reactive protein(CRP)and tumor necrosis factor-alpha(TNF-a)were detected by enzyme-linked immunosorbent assay(ELISA).HUVECs were exposed to different concentrations of LBP before and after LPS induction.Cellular viability was detected by CCK8 for screening the optimal dose of LBP.HUVECs were divided into 5 groups of control,model,LBP,AKT inhibitor and AKT inhibitor plus LBP.Cellular viability was detected by CCK8,apoptosis rate and ROS level were detected by flow cytometry,intracellular MDA content was detected by biochemical kit and protein expressions of AKT,p-AKT,eNOS,p-eNOS and VE-cadherin were detected by Western blot.RESULTS As com-pared with control group,cellular viability of model group declined significantly(P＜0.01),the apoptosis rate was significantly increased(P＜0.01),reactive oxygen species(ROS)level and malondialdehyde(MDA)content spiked markedly(P＜0.01)while intracellular phosphorylation levels of AKT and eNOS protein and expression level of VE-cadherin protein dropped obvi-ously(P＜0.01).As compared with model group,LBP significantly boosted cellular viability(P＜0.01),arrested cell apoptosis(P＜0.01),decreased intracellular ROS level and MDA content(P＜0.01),promoted the phosphorylation of AKT and eNOS(P＜0.01)and up-regulated the expression of VE-cadherin protein(P＜0.01)while AKT inhibitor group displayed the opposite trend.As compared with AKT inhibitor group,LBP could reverse the above effects of AKT inhibitor(P＜0.01).CONCLUSION LBP may alleviate LPS-induced endothelial dysfunction and suppress oxidative stress response and apoptosis.And the underlying mechanism is probably correlated with promoting an activation of AKT/eNOS pathway.
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