Inhibition Effect And Related Mechanism Of Curcumin On Advanced Glycation End Products-induced Expression Of Tumor Necrosis Factor-α And Matrix Metalloproteinase-13 In Rabbit Chondrocytes

AIM In the present study, we used the preparation AGEs as a damage factor to explore the inhibition effects and mechanisms of Curcumin on the AGEs-induced expression of TNF-a and MMP-13 on the cultured rabbit chondrocytes.METHODS The primary cultured rabbit chondrocytes were used in the present study. (1) To explore the effect and the related mechanism of AGEs with different concentration on the expression of TNF-αand MMP-13 mRNA. The experiments were divided into 7 groups:①control;②BSA (100μg/ml);③AGEs(1μg/ml);④AGEs(10μg/ml);⑤AGEs(25μg/ml);⑥AGEs(50μg/ml);⑦AGEs(100μg/ml). The expression of TNF-αand MMP-13 mRNA were designed by RT-PCR and the activity of CAT,SOD and the level of MDA were designed using assay kits. (2) Ivestigating the effect of Curcumin on the AGEs-induced expression of TNF-a and MMP-13, and explore the related mechanism. The experiments were divided into 7 groups:①con;②BSA (100μg/ml);③AGEs(100μg/ml);④AGEs(100μg/ml)+curcumin (10μmol/L);⑤AGEs(100μg/ml)+curcumin(25μmol/L);⑥AGEs(100μg/ml)+curcumin(50μmol/L);⑦curcumin(50μmol/L); Using RT-PCR to detect the mRNA expression of TNF-αand MMP-13and the level of ROS and the activation of NF-κB were designed by specific Fluorescent probe.RESULTS (1) The chondrocytes were treated with AGEs (1, 10,25,50, 100μg/ml) for 48h, and the results showed that AGEs dose-dependently induced TNF-αand MMP-13 mRNA expression (P <0.05 or P<0.01) and the maximum stimulation of chondrocytes were found to be at 100μg/ml AGEs; (2) Both anti-RAGE (5μg/ml) and PDTC(0.1mmol/L) can significantly inhibit the AGEs (100μg/ml)-induced expression of TNF-a and MMP-13 (P<0.01), however, there are no significant different between the groups of treated with anti-RAGE (5μg/ml) or PDTC(0.1mmol/L) alone with the control(P>0.05); (3) After pretreated for 2h,10,25,50μmol/L Curcumin can dose-dependently inhibit TNF-a and MMP-13 expression induced by AGEs (P<0.05 or P<0.01), and the maximum effects were found to be at 50μmol/L Curcumin. The TNF-a/GAPDH OD value decreased from 1.38±0.04 to 0.61±0.02(P<0.01); MMP-13/GAPDH OD value decreased from 0.79±0.04 to 0.31±0.02 (P<0.01) compared with the AGEs (100μg/ml) treated group; and there were no significant different between the 50μmol/L Curcumin alone treated group and the control; (4) AGEs (1,10,25,50,100μg/ml) dose-dependently decreased the activity of CAT and SOD, increased the content of MDA (P<0.05 or P<0.01) incubated with the chondrocytes for 48h; after pretreated with 10,25,50μmol/L Curcumin for 2h, Curcumin concentration-dependently improved the activity of CAT and SOD and decreased the content of MDA compared with the AGEs treatment group (P<0.05 or P<0.01); The maximum effect was found at 50μmol/LCurcumin, the activity of CAT increased from 2.36±0.31 to 11.15±0.52 (P<0.01), the activity of SOD increased from 6.88±0.48 to 22.01±0.62 (P<0.01), the level of MDA decreased from 1.98±0.07 to 0.66±0.0 (P<0.01) compared with AGEs 100μg/ml treatment group; (5) 10,25,50μmol/L Curcumin concentration-dependently inhibit the production of ROS induced by AGEs; the maximum effect was found at 50μmol/LCurcumin, the OD value decreased from 0.39±0.009 to 0.13±0.005 (P<0.01); Curcumin can significantly inhibit the activation of NF-κB induced by AGEs.CONCLUSION 1 AGEs can stimulate the TNF-a and MMP-13 high-expression on the chondrocytes, thus damage the normal function of cells; 2 AGEs can induce the expression of TNF-a and MMP-13 via a mechanism involving the activation of RAGE, the production of ROS and the activation of the signaling pathway of NF-κB; 3 Curcumin can concentration-dependently inhibit the AGEs-induced expression of TNF-a and MMP-13; 4 Curcumin can protect the chondrocytes damage induced by AGEs via a mechanism of inhibiting the RAGE/ROS/NF-κB signaling pathway, thus plays an important role in the prevention and treatment of OA.