Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University, Jinju 660-702.
Department of Pharmacology, School of Medicine, Chungnam National University, Daejeon 301-740, Republic of Korea.
Biomol Ther (Seoul). 2014 May;22(3):239-45. doi: 10.4062/biomolther.2014.020.
We investigated whether luteolin affects the gene expression, secretion and activity of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as production of MMP-3 in the rat knee to evaluate the potential chondro-protective effects of luteolin. Rabbit articular chondrocytes were cultured in a monolayer and IL-1β-induced gene expression levels of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen were measured by reverse transcription - polymerase chain reaction (RT-PCR). Effects of luteolin on interleukin-1β (IL-1β)-induced secretion and enzyme activity of MMP-3 in rabbit articular chondrocytes were investigated by western blot analysis and casein zymography, respectively. The effect of luteolin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) luteolin inhibited the gene expression levels of MMP-3, MMP-1, MMP-13, ADAMTS-4 and ADAMTS-5. However, it increased the gene expression level of collagen in rabbit articular chondrocytes; (2) luteolin inhibited the secretion and activity of MMP-3; (3) luteolin inhibited in vivo production of MMP-3 protein. These results suggest that luteolin can regulate the gene expression, secretion and activity of MMP-3, by directly acting on articular chondrocytes.
我们研究了木樨草素是否影响原代培养兔关节软骨细胞基质金属蛋白酶-3（MMP-3）的基因表达、分泌和活性，以及大鼠膝关节中 MMP-3 的产生，以评估木樨草素的潜在软骨保护作用。兔关节软骨细胞在单层中培养，并通过逆转录-聚合酶链反应（RT-PCR）测量 MMP-3、MMP-1、MMP-13、解整合素金属蛋白酶与凝血酶反应蛋白-4（ADAMTS-4）、ADAMTS-5 和 II 型胶原的基因表达水平。通过 Western blot 分析和酪蛋白酶谱法分别研究了木樨草素对兔关节软骨细胞中白细胞介素-1β（IL-1β）诱导的 MMP-3 分泌和酶活性的影响。还在体内检查了木樨草素对 MMP-3 蛋白产生的影响。结果如下：（1）木樨草素抑制 MMP-3、MMP-1、MMP-13、ADAMTS-4 和 ADAMTS-5 的基因表达水平。然而，它增加了兔关节软骨细胞中胶原蛋白的基因表达水平；（2）木樨草素抑制 MMP-3 的分泌和活性；（3）木樨草素抑制 MMP-3 蛋白的体内产生。这些结果表明，木樨草素可以通过直接作用于关节软骨细胞来调节 MMP-3 的基因表达、分泌和活性。
