Kang Dong-Geun, Lee Hyun Jae, Kim Kun Tae, Hwang Sun-Chul, Lee Choong Jae, Park Jin Sung
Department of Orthopedic Surgery and Institute of Health Sciences, School of Medicine and Hospital, Gyeongsang National University, Changwon 51472, Korea.
Department of Health Management, Sahmyook University, Seoul 01795, Korea.
Korean J Physiol Pharmacol. 2017 Mar;21(2):197-204. doi: 10.4196/kjpp.2017.21.2.197. Epub 2017 Feb 21.
In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-1β-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein . These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.
在本研究中，我们试图检测齐墩果酸是否能调节原代培养的兔关节软骨细胞中基质金属蛋白酶-3（MMP-3）的活性、分泌及基因表达，以及大鼠膝关节中MMP-3的产生，以评估齐墩果酸潜在的软骨保护作用。兔关节软骨细胞进行单层培养，采用逆转录-聚合酶链反应（RT-PCR）检测白细胞介素-1β（IL-1β）诱导的MMP-3、MMP-1、MMP-13、含血小板反应蛋白基序的解聚素和金属蛋白酶-4（ADAMTS-4）、ADAMTS-5及Ⅱ型胶原的基因表达。在兔关节软骨细胞中，分别采用蛋白质印迹分析和酪蛋白酶谱法研究齐墩果酸对IL-1β诱导的MMP-3分泌及蛋白水解活性的影响。在大鼠膝关节内注射齐墩果酸后，还检测了其对MMP-3蛋白产生的影响。结果如下：（1）齐墩果酸抑制MMP-3、MMP-1、MMP-13、ADAMTS-4和ADAMTS-5的基因表达，但增加Ⅱ型胶原的基因表达；（2）齐墩果酸降低MMP-3的分泌及蛋白水解活性；（3）齐墩果酸抑制MMP-3蛋白的产生。这些结果表明，齐墩果酸可通过直接作用于关节软骨细胞来调节MMP-3的活性、分泌及基因表达。
