厚朴酚对骨关节炎的抗炎和软骨保护作用

Anti-inflammatory and cartilage protective effects of magnolol against osteoarthritis

目的探讨厚朴酚的体外抗炎活性和对碘乙酸单钠(MIA)诱导的骨关节炎(OA)的影响。方法脂多糖(LPS)处理的RAW264.7细胞中体外评价厚朴酚的抗炎作用,设立LPS组、厚朴酚不同浓度组(5、10、20μmol/L),以正常培养的细胞作为对照组,Griess试剂测量培养基中NO的累积;Western blot检测细胞iNOS、COX-2蛋白的表达。MIA诱导OA的SD大鼠模型,分为MIA组、厚朴酚不同浓度组(5、10、20 mg/kg),以正常大鼠作为对照组,取大鼠膝关节组织样本进行组织学分析,实时PCR分析大鼠IL-1β、TNF-α、IL-6、MMP-2、MMP-9和COX-2的mRNA表达;ELISA检测培养基上清液中PGE 2、IL-6及各组大鼠血清IL-1β、IL-6和TNF-α含量。结果厚朴酚抑制LPS处理的RAW264.7细胞中的NO、PGE2和IL-6的产生(P<0.05)。此外,厚朴酚抑制MIA诱导OA大鼠模型的IL-1β、TNF-α和IL-6的升高及MMP-2、MMP-9和COX-2的合成(P<0.05)。结论厚朴酚在OA大鼠模型中发挥有效的抗炎活性并保护软骨。

This study was designed to investigate the in vitro anti-inflammatory activity of magnolol and its effect on osteoarthritis(OA) induced by monoethyl iodide(MIA). The in vitro anti-inflammatory effect of magnolol was evaluated by lipopolysaccharide(LPS)-treated RAW264.7 cells. LPS group and magnolol groups(5, 10,20 μmol/L) were set up, and the normal cultured cells were used as the control group. Griess reagent was used to measure the accumulation of NO in the medium;the expression of iNOS and COX-2 proteins were detected by Western blotting. The SD rat model of MIA-induced OA was divided into the MIA group and magnolol groups(5, 10,20 mg/kg), while normal rats were used as control. the rat knee joint tissue samples were taken from all groups for histology;Real-time PCR was used to analyze the mRNA expression of IL-1β, TNF-α, IL-6, MMP-2, MMP-9 and COX-2 in rats of different groups;while ELISA was employed to detect the levels of PGE 2, IL-6 in medium supernatant, and the serum levels of IL-1β, IL-6 and TNF-α in rats. Data showed that magnolol inhibited the production of NO, PGE2 and IL-6 in LPS-treated RAW264.7 cells(P<0.05). In addition, magnolol inhibited the increase of IL-1β, TNF-α and IL-6 and the synthesis of MMP-2, MMP-9 and COX-2 in MIA-induced OA rat model(P<0.05). Therefore, magnolol exerts potent anti-inflammatory activity and protects cartilage in the OA rat model.