Effect and mechanism of carnosic acid on interleukin-1β induced degradation of chondrocyte extracellular matrix
Objective:To investigate the effect and mechanism of carnosic acid(CAD)on interleukin-1β(IL-1β)induced degradation of chondrocyte extracellular matrix.Methods:Primary rat chondrocytes were cultured in vitro and randomly divided into four groups:control group,IL-1β group(10 ng/mL),CAD(4 μmol/L)+IL-1β group,and CAD(8 μmol/L)+IL-1β group.After 24 hours of treatment,a CCK-8 kit was used to detect the effect of different concentrations of CAD(0,1,2,4,8,16,32,and 64 μmol/L)on chondrocyte proliferation.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of tumor necrosis factor-α(TNF-α),matrix metalloproteinase 13(MMP13),a disintegrin and metalloproteinase with thrombospondin 5(ADAMTS5),SRY-box tran-scription factor 9(Sox9),collagen type Ⅱ alpha 1 chain(Col2a1)and aggrecan(ACAN)mRNA in chon-drocytes.MMP13,Col2a1,and P65 protein expression levels in chondrocytes were detected by im-munofluorescence.Western Blot was used to detect the protein expression and quantitative analysis of P65 in chondrocytes.Molecular docking was used to verify the potential targets of CAD.Results:Com-pared with the control group,high concentrations of CAD(16,32,and 64 μmol/L)inhibited the proliferation of chondrocytes in a concentration-dependent manner.Moreover,compared with the con-trol group,IL-1β significantly promoted the expression of TNF-α,MMP13 and ADAMTS5,and inhib-ited the expression of Sox9,Col2a1 and ACAN in chondrocytes.However,CAD(4 and 8 μmol/L)could significantly ameliorate the matrix degradation of chondrocytes induced by IL-1β(P<0.05).At the mechanism level,CAD could significantly inhibit the nuclear translocation of P65 induced by IL-1β,and molecular docking showed that CAD had a good binding to P65.Conclusion:CAD could signifi-cantly reverse the matrix degradation of chondrocytes induced by IL-1β and inhibit the nuclear factor-kap-pa B(NF-κB)signal pathway through P65.
NETL
NSTL
万方数据
