机体炎症因子和氧化应激标志物介导姜黄素抑制骨性关节炎的作用机制

目的：探讨骨性关节炎（OA）关节软骨基质金属蛋白酶-2（MMP-2）、单核细胞趋化蛋白-1（MCP-1）、CD47、L-选择素及高级氧化蛋白产物（AOPP）表达的变化以及姜黄素（Cur）的干预作用。方法：建立C57BL/6小鼠OA模型，20只小鼠随机分为手术对照组、模型对照组、Cur25组和Cur50组（术后分别每日接受25 μmol/L和50 μmol/L浓度姜黄素腹腔注射）。4周后取材，采用免疫组织化学法观察各组大体标本形态学变化，电子显微镜下观察软骨组织超微结构；培养各组软骨细胞，运用蛋白质印迹法检测各组MMP-2、MCP-1和CD47表达，ELISA和分光光度法检测L-选择素水平及AOPP浓度变化。结果：软骨组织标本形态学变化模型对照组最明显，Cur25组次之；与手术对照组比较，模型对照组、Cur25组、Cur50组软骨细胞MMP-2、MCP-1表达均增多（均P<0.05），CD47表达均下降（均P<0.05），L-选择素水平及AOPP浓度均增加（均P<0.05）；与模型对照组比较，Cur25组和Cur50组软骨细胞MMP-2、MCP-1、L-选择素及AOPP的表达下降（均P<0.05），CD47的表达上升（均P<0.05），且与Cur25组比较，Cur50组的上述变化更为显著（均P<0.05）。结论：MMP-2、MCP-1、CD47、L-选择素及AOPP的异常表达与OA软骨退变的病理过程相关。姜黄素可缓解关节软骨的退变，减轻软骨炎症，提高软骨细胞的代谢活性，且此作用与姜黄素的浓度有关。
Abstract: Objective: To observe the influence of matrix metalloproteinases-2 (MMP-2), monocyte chemoattractant protein-1 (MCP-1), CD47, L-selectin and advanced oxidation proteinproducts (AOPP) in osteoarthritis and the intervention of curcumin. Methods: A total of 20 male C57BL/6 mice (10.05-15.00 g) were randomly divided into control group, OA group, Cur25 group and Cur50 group (intraperitoneal injected 25 μmol/L or 50 μmol/L of curcumin everyday after modeling). After 4 weeks treatment, we observed the morphological changes of the gross specimen by immunohistochemical method, and observed the ultrastructure of cartilage tissue under electron microscope. The expression of MMP-2, MCP-1 and CD47 were detected by western blotting, and L-selectin and AOPP were detected by ELISA and spectrophotometer, respectively. Results: In the cartilage tissue morphology, the chondrocytes of OA group showed obvious change, while Cur25 and Cur50 groups maintained the good cartilage cell membrane intact. Compared with control group, the expressions of MMP-2, MCP-1, L-selectin and AOPP in OA group, Cur25 group and Cur50 group were increased (all P<0.05), while CD47 levels were decreased (all P<0.05). Compared with OA group, the expressions of MMP-2, MCP-1, L-selectin and AOPP in Cur25 group and Cur50 group were decreased (all P<0.05), while CD47 levels were increased (all P<0.05), and such changes were more significant in Cur50 group (all P<0.05). Conclusion: The MMP-2, MCP-1, CD47, L-selectin and AOPP are closely associated with the pathology course of OA. Curcumin has protection effect on cartilage, which can relieve joint cartilage degeneration, reduce cartilage inflammation and increase the metabolic activity of chondrocytes. Key words: Osteoarthritis/pathology Curcumin/pharmacology Curcumin/administration & dosage Matrix metalloproteinase 2 Chemokine CCL2 Antigens, CD47