姜黄素通过上调PPARγ抑制糖基化终末产物诱导软骨细胞分泌TNF-α及MMP-13  

Inhibition effect of curcumin on advanced glycation end products-induced expression of tumor necrosis factor-α and matrix metalloproteinase-13 in rabbit chondrocytes by elevating peroxisome proliferators-activated receptor γ

作　　者：杨青山[1] 吴树金[2] 毛新展[3] 陈志信[1] 台会平[1] 

机构地区：[1]甘肃省人民医院骨科,兰州730000 [2]甘肃省人民医院药剂科,兰州730000 [3]中南大学湘雅二医院骨科,长沙410011

出　　处：《中南药学》2017年第12期1678-1682,共5页Central South Pharmacy

基　　金：甘肃省卫生行业科研计划资助项目(编号:GSWSKY-2014-10);甘肃省中医药管理局科研计划项目(编号:GZK-2013-42)

摘　　要：目的在体外培养的家兔软骨细胞模型上,观察姜黄素(curcumin)对糖基化终末产物(AGEs)诱导的软骨细胞TNF-α和MMP-13表达的影响,并探讨其相关机制是否与姜黄素上调PPARγ有关。方法采用real-time PCR方法检测TNF-α、MMP-13、PPARγmRNA的表达量;试剂盒方法检测CAT、SOD活性及MDA水平,荧光探针法检测ROS水平;Western blot检测PPARγ蛋白含量;DNA-binding法检测PPARγ活性。结果 AGEs(100μg·mL^(-1))与软骨细胞共孵育48 h后,TNF-α及MMP-13 mRNA的表达较正常对照组明显升高(P<0.05),50μmol·L^(-1)的姜黄素与软骨细胞预孵育2 h后,可以显著抑制由AGEs诱导的TNF-α及MMP-13 mRNA增多,拮抗由AGEs所致细胞CAT、SOD活性降低及MDA、ROS水平增高(P<0.05);给予PPARγ特异性抑制剂GW9662 10μmol·L^(-1)预处理后,可以显著拮抗姜黄素对AGEs诱导软骨细胞损伤及氧化应激的保护作用;姜黄素显著上调AGEs诱导的PPARγ活性降低,伴随PPARγ相应mRNA及蛋白表达水平的升高。结论姜黄素可能通过上调PPARγ从而降低ROS水平,进而抑制由AGEs诱导的TNF-α和MMP-13表达增多,保护软骨细胞的功能。Objective To determine the inhibition effect of curcumin on the expression of TNF-α and MMP-13 induced by advanced glycation end products （AGEs） through elevating PPARy in cultured rabbit chondrocytes. Methods The expression of TNF-α, MMP-13 and PPARγ mRNA were measured by RT-PCR; the activity of CAT, and SOD and the level of MDA were measured by assay kits, and the level of ROS by specific fluorescent probe. PPARy protein was determined by Western blot and the activity of PPARy was assayed by DNA-binding method. Results After the chondrocytes were treated with AGEs （100 μg· mL-1） for 48 h, the expression of TNF-α and MMP-13 mRNA was significantly increased as compared with that in the control group （P 〈 0.05）. While 50 μmol· L - 1 curcumin after treating the chondrocytes for 2 h significantly inhibited TNF-α and MMP- 13 expression increase induced by AGEs （P 〈 0.05）, improved the activity of CAT and SOD and decreased the content of MDA and ROS level （P 〈 0.05）. However, blocking of PPARy with GW9662 （10 μmol · L - 1） decreased the protective effect of curcumin against AGEs-induced chondrocyte damage. In addition, curcumin significantly increased the PPARy transcriptional activity induced by AGEs, together with a significant increase in PPARy protein and mRNA expression. Conclusion Curcumin can alleviate AGEs-induced chondrocyte damage by the modulation of PPARy and inhibit the ROS level.

关 键 词：糖基化终末产物 姜黄素 软骨细胞 过氧化物酶体增殖物激活受体Γ 

分 类 号：R285[医药卫生—中药学]