Kang Dong-Geun, Lee Hyun Jae, Lee Choong Jae, Park Jin Sung
Department of Orthopaedic Surgery, Gyeongsang National University Changwon Hospital, Gyeongsang National University School of Medicine, Changwon 51472, Republic of Korea.
Smith Liberal Arts College and Department of Addiction Science, Graduate School, Sahmyook University, Seoul 01795, Republic of Korea.
Biomol Ther (Seoul). 2018 Nov 1;26(6):560-567. doi: 10.4062/biomolther.2018.132.
In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B (NF-κB) in articular chondrocytes. 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. Effect of resveratrol on IL-1β-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on IL-1β-induced NF-κB signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol 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) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited IL-1β-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Bα (IκBα); (4) resveratrol inhibited IL-1β-induced phosphorylation and nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting NF-κB by directly acting on articular chondrocytes.
在本研究中，我们试图检测白藜芦醇是否通过影响关节软骨细胞中的核因子-κB（NF-κB）来调节基质金属蛋白酶（MMPs）的表达。将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。采用蛋白质印迹分析研究白藜芦醇对兔关节软骨细胞中IL-1β诱导的MMP-3分泌的影响。为阐明白藜芦醇的作用机制，通过蛋白质印迹分析在人软骨肉瘤细胞系SW1353中研究白藜芦醇对IL-1β诱导的NF-κB信号通路的影响。结果如下：（1）白藜芦醇抑制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）白藜芦醇减少MMP-3的分泌；（3）白藜芦醇抑制IL-1β诱导的抑制性κB激酶（IKK）激活（磷酸化），进而抑制抑制性κBα（IκBα）的磷酸化和降解；（4）白藜芦醇抑制IL-1β诱导的NF-κB p65磷酸化和核转位。这进而导致SW1353细胞中MMPs基因表达下调。这些结果表明，白藜芦醇可通过直接作用于关节软骨细胞影响NF-κB来调节MMPs的表达。
