Qin Jun, Shang Liang, Ping An-song, Li Jing, Li Xiao-jun, Yu Hong, Magdalou Jacques, Chen Liao-bin, Wang Hui
Arthritis Res Ther. 2012 Nov 7;14(6):R242. doi: 10.1186/ar4085.
Sodium ferulate (SF) is a natural component of traditional Chinese herbs. Our previous study shows that SF has a protective effect on osteoarthritis (OA). The objective of this study was to investigate the effect of SF on the TNF/TNF receptor (TNFR) signal transduction pathway of rat OA chondrocytes.
Primary rat articular chondrocytes were co-treated with IL-1β and SF. Chondrocyte apoptosis was assessed by fluorescein isothiocyanate-annexin V/propidium iodide assay. The PCR array was used to screen the expression of 84 key genes involved in apoptosis. The release of TNFα and prostaglandin E2 were analyzed by ELISA. Expressions of proteins were assessed by western blotting. The activity of NF-κB was determined by electrophoretic mobility shift assay (EMSA). Gene expression of inducible nitric oxide synthase (iNOS) was evaluated by real-time quantitative PCR. The nitric oxide content was measured with the Griess method.
After treatment with SF, the apoptosis rate of chondrocytes significantly attenuated (P < 0.01). Results of the apoptosis PCR array suggested that mRNA expression of some core proteins in the TNF/TNFR pathway showed valuable regulation. The protein expressions of TNFα, TNFR-1, TNF receptor-associated death domain, caspase-8 and caspase-3 were prevented by SF in a concentration-dependent manner. SF also inhibited activities of caspase-8 and caspase-3 compared with the OA model control (P < 0.01). TNF receptor-associated factor-2 expression, phosphorylations of inhibitor of NF-κB kinase (IKK) subunits alpha and beta, and NF-κB inhibitor, alpha (IκBα) were all concentration-dependently suppressed by SF treatment. The results of EMSA showed that SF inhibited the activity of NF-κB. In addition, the expressions of cycloxygenase-2 and iNOS and the contents of prostaglandin E2 and NO were attenuated with the treatment of SF (P < 0.01).
SF has anti-apoptosis and anti-inflammatory effects on an OA model induced by IL-1β in vitro, which were due to inhibitory actions on the caspase-dependent apoptosis pathway and the IKK/NF-κB signal transduction pathway of the TNF/TNFR pathway.
阿魏酸钠（SF）是传统中草药的天然成分。我们之前的研究表明，SF对骨关节炎（OA）具有保护作用。本研究的目的是探讨SF对大鼠OA软骨细胞肿瘤坏死因子/肿瘤坏死因子受体（TNFR）信号转导通路的影响。
将原代大鼠关节软骨细胞与白细胞介素-1β（IL-1β）和SF共同处理。通过异硫氰酸荧光素-膜联蛋白V/碘化丙啶检测评估软骨细胞凋亡。采用PCR芯片筛选84个参与凋亡的关键基因的表达。通过酶联免疫吸附测定（ELISA）分析肿瘤坏死因子α（TNFα）和前列腺素E2的释放。通过蛋白质免疫印迹法评估蛋白质表达。通过电泳迁移率变动分析（EMSA）测定核因子κB（NF-κB）的活性。通过实时定量PCR评估诱导型一氧化氮合酶（iNOS）的基因表达。用格里斯方法测量一氧化氮含量。
用SF处理后，软骨细胞的凋亡率显著降低（P<0.01）。凋亡PCR芯片结果表明，TNF/TNFR通路中一些核心蛋白的mRNA表达呈现出有价值的调控。SF以浓度依赖性方式抑制TNFα、TNFR-1、肿瘤坏死因子受体相关死亡结构域、半胱天冬酶-8和半胱天冬酶-3的蛋白表达。与OA模型对照组相比，SF还抑制了半胱天冬酶-8和半胱天冬酶-3的活性（P<0.01）。SF处理浓度依赖性地抑制肿瘤坏死因子受体相关因子-2表达、NF-κB抑制蛋白激酶（IKK）亚基α和β的磷酸化以及NF-κB抑制蛋白α（IκBα）。EMSA结果表明，SF抑制了NF-κB的活性。此外，随着SF处理，环氧合酶-2和iNOS的表达以及前列腺素E2和一氧化氮的含量降低（P<0.01）。
SF对体外IL-1β诱导的OA模型具有抗凋亡和抗炎作用，这归因于对TNF/TNFR通路中半胱天冬酶依赖性凋亡途径和IKK/NF-κB信号转导途径的抑制作用。
