双醋瑞因对IL-1β诱导软骨细胞损伤的保护作用

Protective Effect of Diacerein on Interleukin-1β-induced Rat Chondrocytes Injury

关键词:  双醋瑞因  骨关节炎  软骨细胞  炎症

Key words:  diacerein  ostearthritis  chondrocytes  inflammation

摘要:

目的 探讨双醋瑞因对白介素-1β(IL-1β)诱导软骨细胞损伤的保护作用。方法 通过Ⅱ型胶原酶消化SD大鼠软骨，获得软骨细胞，10 ng·L^-1 IL-1β诱导软骨损伤48 h的同时，1，10，100 μmol·L^-1双醋瑞因也干预48 h。MTT法检测双醋瑞因对IL-1β诱导的骨关节炎软骨细胞活力的修复作用；Griess法检测IL-1β诱导的骨关节炎软骨细胞上清NO水平；Western blot分析炎症因子环氧化酶-2(cyclooxygenase-2，COX-2)，核因子-κB (NF-κB)信号通路相关蛋白p65，pp65的含量变化。结果 MTT法显示双醋瑞因(1，10，100 μmol·L^-1)可改善IL-1β诱导的大鼠软骨细胞的活力，且呈现剂量依赖关系。Griess法检测结果表明双醋瑞因能抑制IL-1β诱导的骨关节炎软骨细胞上清NO含量上升。Western blot结果显示双醋瑞因能抑制IL-1β诱导的骨关节炎软骨细胞炎症因子COX-2含量上升；并且抑制核因子p65磷酸化。结论 双醋瑞因能够抑制IL-1β诱导的软骨细胞炎症，可能是通过NF-κB信号通路来实现的。

Abstract:

OBJECTIVE To investigate the protective effect of diacerein on interleukin-1β-induced rat chondrocytes injury. METHODS Chondrocytes were isolated from the knee joints of Sprague-Dawley rats by enzymatic digestion. Rat chondrocytes were co-treated with IL-1β(10 ng·mL^-1) and diacerein(1, 10, 100 μmol·L^-1) for 48 h. The viability of chondrocytes was detected by MTT assay. The level of NO was assayed by Griess method. Inflammation related protein cyclooxygenase-2(COX-2) was assayed by Western blot. Finally, the expression of nuclear factor kappa B(NF-κB) p65 and p65 phosphorylation were detected by Western blot. RESULTS MTT assay demonstrated diacerein(1, 10, 100 μmol·L^-1) could protect IL-1β-induced chondrocytes viability. Diacerein also inhibited the increasing levels of NO in IL-1β-induced chondrocytes. Western blot assays presented diacerein could suppress IL-1β-induced increasing levels of COX-2. What’s more, diacerein inhibited NF-κB p65 phosphorylation induced by IL-1β in rat chondrocytes. CONCLUSION Diacerein exertes anti- inflammation in IL-1β-induced rat chondrocytes, which might be related to NF-κB signal pathway.