Hsu H-C, Ke Y-L, Lai Y-H, Hsieh W-C, Lin C-H, Huang S-S, Peng J-Y, Chen C-H
Ching-Hsein Chen, Professor, Department of Microbiology, Immunology and Biopharmaceuticals, College of Life Sciences, National Chiayi University, Chiayi City 60004, Taiwan, ROC. E-mail:
J Nutr Health Aging. 2022;26(3):307-313. doi: 10.1007/s12603-022-1752-5.
Chondroitin sulfate (CS) is found in humans' cartilage, bone, cornea, skin, and arterial wall. It consists of the foundation substance in the extracellular matrix (ECM) of connective tissue. The oral supplement form of CS is clinically used in treating osteoarthritis (OA).
Cell migration was observed by the transwell assay. The EMT, Akt/IKK/IκB pathways, TIMPs, collagen and MMPs in cell lysate were determined by Western blotting. The expression of MMP activity was determined by gelatin zymography. The production of reactive oxygen species (ROS) was determined by using a fluorescence spectrophotometer.
In the current report, we demonstrated that CS can increase the cell proliferation and migration of chon-001 chondrocytes. Treatment with CS induced the epithelial-mesenchymal transition and increased the expression of type II collagen and TIMP-1/TIMP2 and inhibited the expressions and activities of metalloproteinase-9 (MMP-9) and metalloproteinase-2 (MMP-2). The phosphorylation of Akt, IκB kinase (IKK), IκB and p65 was decreased by CS. CS treatment resulted in β-catenin production and XAV939, a β-catenin inhibitor, and inhibited the cell proliferation by CS treatment. In addition, also significantly induced intracellular ROS generation. Treatment with antioxidant propyl gallate blocked cell migration induced by CS.
We demonstrated that CS induced cell proliferation and migration of chondrocytes by inducing β-catenin and enhancing ROS production. Moreover, our studies demonstrated that CS can increase the activity of chondrocytes and help patients with osteoarthritis to restore cartilage function.
硫酸软骨素(CS)存在于人体的软骨、骨骼、角膜、皮肤和动脉壁中。它是结缔组织细胞外基质(ECM)中的基础物质。CS的口服补充剂形式在临床上用于治疗骨关节炎(OA)。
通过Transwell实验观察细胞迁移。通过蛋白质免疫印迹法测定细胞裂解物中的上皮-间质转化、Akt/IKK/IκB信号通路、组织金属蛋白酶抑制剂(TIMPs)、胶原蛋白和基质金属蛋白酶(MMPs)。通过明胶酶谱法测定MMP活性的表达。使用荧光分光光度计测定活性氧(ROS)的产生。
在本报告中,我们证明CS可以增加chond-001软骨细胞的增殖和迁移。CS处理诱导上皮-间质转化,增加II型胶原蛋白和TIMP-1/TIMP2的表达,并抑制基质金属蛋白酶-9(MMP-9)和基质金属蛋白酶-2(MMP-2)的表达和活性。CS降低了Akt、IκB激酶(IKK)、IκB和p65的磷酸化水平。CS处理导致β-连环蛋白产生,而β-连环蛋白抑制剂XAV939抑制了CS处理诱导的细胞增殖。此外,CS还显著诱导细胞内ROS生成。用抗氧化剂没食子酸丙酯处理可阻断CS诱导的细胞迁移。
我们证明CS通过诱导β-连环蛋白和增强ROS产生来诱导软骨细胞的增殖和迁移。此外,我们的研究表明CS可以增加软骨细胞的活性,并帮助骨关节炎患者恢复软骨功能。