| ObjectiveTo observe the protective effect of lycopene on TNF-α-induction endothelial cellinjury, and then explore its further pharmacologic actions, and provide reliableexperiment evidence for clinical application.MethodsHUVEC cells were cultured in vitro, and then were divided into5groups according tothe protocols:(1)Negative control (group A);(2) TNF-α (group B,50ng/mL TNF-α);(3)low-dose of lycopene (group C,1μmol/L lycopene for24h and then50ng/mL ofTNF-α was added);(4) middle dose group (group D,50ng/mL of TNF-α with5μmol/L lycopene);(5) middle dose group (group E,50ng/mL of TNF-α with10μmol/L lycopene). After treatment, cells proliferation was detected by WST-1;secretion of MMP-9was determined by ELISA; the activity of MMP-9was measuredby gelatinase experiment. Cytosolic and nuclear extracts were prepared, and NF-κBp65and phosphorylated I-κB were detected by western blot.Results(1)0～10μmol/L lycopene was not toxic to HUVEC cells. When then concentrationwas reached to15μmol/L, the cells proliferation rates was reduced by7%;(2) When HUVEC cells were stimulated by50ng/mL TNF-α for24h, the cells growthsuppression rate was32%; however, when HUVEC cells were pretreated by1~10μmol/L lycopene, the Growth suppression rate was dropped to20.4%,14.6%, and4.51%, respectively;(3) After HUVEC cells were treated by TNF-α, the concentration of MMP-9was(1.17±0.24) ng/ml, and1~10μmol/L lycopene inhibited the expression and activity of MMP-9in dose-dependence manner;(4) NF-κB p65subunit was not detected in normal group; when the cells were treatedby TNF-α for24h, p65subunit was increased in the nuclear.1~10μmol/Llycopene could inhibit the p65subunit expression significantly;(5) Phosphorylated I-κB was low in cytoplasm when it was unstipulated by TNF-α.Lycopene could inhibit the increase of phosphorylated I-κB.Conclusions(1) Lycopene could suppress the MMP-9expression in HUVEC cells caused byTNF-α.(2) Lycopene inhibit the expression of MMP-9may be associated with NF-κBactivation... |
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