Effect Of Resveratrol On IL-1β Induced Inflammatory Mediators In Human Nucleus Pulposus Cells

ObjectiveTo investigate the influence of Resveratrol on secretion of IL-6, IL-8, and expressionof matrix metalloproteinase (MMP)-3as well as tissue inhibitor ofmetalloproteinase-1(TIMP)-1in human nucleus pulposus cells, and then elucidatethe possible mechanism involved in it.MethodsThe primary nucleus pulposus cells were isolated, and divided into three groupsaccording to the experimental purposes:(1) Control group (Equal volume of dimethylsulfoxide was added);(2) IL-1β group (Cells were treated with5ng/ml IL-1β for2h);(3) Resveratrol group（Cells were stimulated with5μmol/L,30μmol/L,50μmol/LResveratrol for0h,12h,24h,36h or48h. Production of IL-6and IL-6were detectedby ELISA, expression of MMP-3and TIMP-1mRNA were determined by real-timePCR, and nuclear translocation of NF-κB p65subunit was analyzed by Western blot.Results1. ELISA results show that the secretion of IL-6and IL-8was very low in controlgroup. IL-1β treatment significantly increased production of IL-6and IL-8, andpeaks at24h. However, they were significantly decreased when treated by5μmol/L,30μmol/L and50μmol/L of Resveratrol.2. Real time PCR showed expression of few MMP-3and TIMP-1mRNA wereexpressed. IL-1β treatment increase MMP-3, and decreased TIMP-1to50%.Different concentration of Resveratrol incubation decreased MMP-3and increased TIMP-1to normal level.3Western blot evidences that NF-κB p65subunit significantly translocate intonucleus after IL-1β stimulation. Similarly, p65translocation was reduced asthe concentration of Resveratrol increased.Conclusion1. Resveratrol can inhibit IL-1β-induced secretion of IL-6and IL-8, and MMP-3expression, increase TIMP-1expression.2. The inhibit effect of Resveratrol may achieve by inhibition of NF-κB...