姜黄素对白细胞介素1β诱导兔骨关节炎标志物的作用  

Effects of curcumin on rabbit osteoarthritis markers induced by interleukin-1 beta

作　　者：王健[1] 王祥[1] 马捷[1] 岳冰[1] 尚修帅 陶海荣[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院骨科,201999

出　　处：《中华关节外科杂志（电子版）》2016年第3期63-68,共6页Chinese Journal of Joint Surgery(Electronic Edition)

基　　金：上海市卫生和计划生育委员会课题(201540274)

摘　　要：目的探讨姜黄素对白细胞介素1β(IL-1β)体外诱导兔关节软骨细胞分泌基质金属蛋白酶-13(MMP-13)、环氧化酶-2(COX-2)、Ⅱ型胶原的影响及可能涉及的信号通路转导。方法分离兔软骨细胞进行体外培养及鉴定,用IL-1β不同时间点刺激软骨细胞,Western Blot检测MMP-13、COX-2的表达。将IL-1β分别与不同浓度的姜黄素共育于兔关节软骨细胞不同时间点,Western Blot检测MMP-13和Ⅱ型胶原的表达,以期获得最佳的作用浓度和时间。使用姜黄素和(或)特异性的p38丝裂原活化蛋白激酶(MAPK)抑制剂SB203580共培养IL-1β刺激的软骨细胞,RT-q PCR检测细胞MMP-13、COX-2、Ⅱ型胶原;Western Blot检测p38、p-p38的表达。结果体外分离培养软骨细胞并鉴定,IL-1β上调软骨细胞的MMP-13、COX-2的表达,姜黄素能够抑制p38的磷酸化,p-p38表达有统计学意义(t=11.22,P<0.01);姜黄素下调IL-1β刺激软骨细胞的MMP-13、COX-2的表达,同时上调Ⅱ型胶原的表达,与IL-1β组相比差异有统计学意义(tMMP=15.79,tCOX=10.27,tⅡ胶原=5.00,P值均小于0.01)。结论姜黄素能够通过减少p38的磷酸化抑制p38 MAPK途径,显著抑制IL-1β刺激兔软骨细胞分泌MMP-13和COX-2,减少软骨基质的降解,增加Ⅱ型胶原的含量,延缓软骨退变,并对软骨细胞具有一定的保护功能。Objective To investigate the effects of curcumin on the secretion of metalloproteinase-13（ MMP-13）,cyclooxygenase-2（ COX-2） and type Ⅱ collagen induced by interleukin-1 beta（ IL-1β）in vitro,and explore the possible signaling pathway involved in this process. Methods Isolated rabbit chondrocytes were cultured and identified; the cartilage cells were stimulated by IL-1β at different time points. Western blot was conducted to evaluate the expression of MMP-13 and COX-2. In order to acquire the optimum time point and concentration of curcumin acting on chondrocytes,rabbit articular chondrocytes were co-treated with IL-1β and curcumin at different concentrations. The chondrocytes were harvested at the indicated time points and the expression of MMP-13 and type Ⅱ collagen was examined by Western Blot analysis. Isolated articular chondrocytes were first treated with IL-1β,and then co-treated with curcumin,or SB203580 [inhibitor of p38 mitogen-activated protein kinases（ MAPK） ],or curcumin plus SB203580,Real time quantitative polymerase chain reaction（ RT-q PCR） was performed to evaluate the expression of MMP-13,COX-2 and type Ⅱ collagen at the mRNA level,and the expression of p38 and p-p38 at the protein level were examined by Western Blot. Results The chondrocytes were cultured and identified in vitro,IL-1β increased MMP-13,COX-2 expression in the cartilage cells,while curcumin inhibited the phosphorylation of p38. Curcumin inhibited the phosphorylation of p38 and p-p38 expression was statistically significant（ t = 11. 22,P〈0. 01）. Compared with IL-1β group,curcumin downregulated IL-1β and stimulated the expression of COX-2 and MMP-13 in chondrocyte; it also upregulated type Ⅱcollagen expression,which were statistically significant（ MMP： t = 15. 79; COX： t = 10. 27; collagen Ⅱ：t = 5. 00; all P〈0. 01）. Conclusions Curcumin inhibits IL-1β-induced secretion of MMP-13 and COX-2in rabbit chondrocytes through suppression of p38 phosphorylation and p38 MAPK signaling pat

关 键 词：姜黄素 骨关节炎 丝裂原活化蛋白激酶 基质金属蛋白酶类 环氧化酶2 

分 类 号：R684.3[医药卫生—骨科学]