Toegel S, Wu S Q, Piana C, Unger F M, Wirth M, Goldring M B, Gabor F, Viernstein H
Department of Pharmaceutical Technology and Biopharmaceutics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
Osteoarthritis Cartilage. 2008 Oct;16(10):1205-12. doi: 10.1016/j.joca.2008.01.013. Epub 2008 Mar 5.
To compare the effects of glucosamine (GlcN), curcumin, and diacerein in immortalized human C-28/I2 chondrocytes at the cellular and the gene expression level. This study aimed to provide insights into the proposed beneficial effects of these agents and to assess the applicability of the C-28/I2 cell line as a model for the evaluation of chondroprotective action.
Interleukin-1beta (IL-1beta)-stimulated C-28/I2 cells were cultured in the presence of GlcN, curcumin, and diacerein prior to the evaluation of parameters such as viability, morphology and proliferation. The impact of GlcN, curcumin, and diacerein on gene expression was determined using quantitative real-time RT-PCR (qPCR).
At the transcriptional level, 5 mM GlcN and 50 microM diacerein increased the expression of cartilage-specific genes such as aggrecan (AGC) and collagen type II (COL2), while reducing collagen type I (COL1) mRNA levels. Moreover, the IL-1beta-mediated shift in gene expression pattern was antagonized by GlcN and diacerein. These effects were associated with a significant reduction in cellular proliferation and the development of chondrocyte-specific cell morphology. In contrast, curcumin was not effective at lower concentrations but even damaged the cells at higher amounts.
Both GlcN and diacerein promoted a differentiated chondrocytic phenotype of immortalized human C-28/I2 chondrocytes by altering proliferation, morphology, and COL2/COL1 mRNA ratios. Moreover, both agents antagonized inhibitory effects of IL-1beta by enhancing AGC and COL2 as well as by reducing COL1 mRNA levels.
在细胞水平和基因表达水平上比较氨基葡萄糖（GlcN）、姜黄素和双醋瑞因对永生化人C-28/I2软骨细胞的影响。本研究旨在深入了解这些药物的潜在有益作用，并评估C-28/I2细胞系作为评估软骨保护作用模型的适用性。
在评估细胞活力、形态和增殖等参数之前，将白细胞介素-1β（IL-1β）刺激的C-28/I2细胞在GlcN、姜黄素和双醋瑞因存在的情况下进行培养。使用定量实时逆转录聚合酶链反应（qPCR）确定GlcN、姜黄素和双醋瑞因对基因表达的影响。
在转录水平上，5 mM GlcN和50 μM双醋瑞因增加了软骨特异性基因如聚集蛋白聚糖（AGC）和II型胶原蛋白（COL2）的表达，同时降低了I型胶原蛋白（COL1）mRNA水平。此外，GlcN和双醋瑞因拮抗了IL-1β介导的基因表达模式变化。这些作用与细胞增殖的显著降低和软骨细胞特异性细胞形态的形成有关。相比之下，姜黄素在较低浓度下无效，但在较高剂量时甚至会损伤细胞。
GlcN和双醋瑞因均通过改变增殖、形态和COL2/COL1 mRNA比率，促进了永生化人C-28/I2软骨细胞的分化软骨细胞表型。此外，这两种药物通过增强AGC和COL2以及降低COL1 mRNA水平，拮抗了IL-1β的抑制作用。
