Tonon R, D'Andrea P
Dipartimento di Biochimica, Biofisica e Chimica delle Macromolecole, Università di Trieste, Italy.
J Bone Miner Res. 2000 Sep;15(9):1669-77. doi: 10.1359/jbmr.2000.15.9.1669.
Cell-to-cell interactions and gap junctions-dependent communication are crucially involved in chondrogenic differentiation, whereas in adult articular cartilage direct intercellular communication occurs mainly among chondrocytes facing the outer cartilage layer. Chondrocytes extracted from adult articular cartilage and grown in primary culture express connexin 43 (Cx43) and form functional gap junctions capable of sustaining the propagation of intercellular Ca2+ waves. Degradation of articular cartilage is a characteristic feature of arthritic diseases and is associated to increased levels of Interleukin-1 (IL-1) in the synovial fluid. We have examined the effects of IL-1 on gap junctional communication in cultured rabbit articular chondrocytes. Incubation with IL-1 potentiated the transmission of intercellular Ca2+ waves and the intercellular transfer of Lucifer yellow. The stimulatory effect was accompanied by a dose-dependent increase in the expression of Cx43 and by an enhanced Cx43 immunostaining at sites of cell-to-cell contact. IL-1 stimulation induced a dose-dependent increase of cytosolic Ca2+ and activates protein tyrosine phosphorylation. IL-1-dependent up-regulation of Cx43 could be prevented by intracellular Ca2+ chelation but not by inhibitors of protein tyrosine kinases, suggesting a crucial role of cytosolic Ca2+ in regulating the expression of Cx43. IL-1 is one of the most potent cytokines that promotes cartilage catabolism; its modulation of intercellular communication represents a novel mechanism by which proinflammatory mediators regulate the activity of cartilage cells.
细胞间相互作用和缝隙连接依赖的通讯在软骨形成分化中至关重要,而在成年关节软骨中,直接的细胞间通讯主要发生在面向软骨外层的软骨细胞之间。从成年关节软骨中提取并在原代培养中生长的软骨细胞表达连接蛋白43(Cx43)并形成能够维持细胞间Ca2+波传播的功能性缝隙连接。关节软骨的降解是关节炎疾病的一个特征,并且与滑液中白细胞介素-1(IL-1)水平的升高有关。我们研究了IL-1对培养的兔关节软骨细胞中缝隙连接通讯的影响。用IL-1孵育可增强细胞间Ca2+波的传递和荧光素黄的细胞间转移。这种刺激作用伴随着Cx43表达的剂量依赖性增加以及细胞间接触部位Cx43免疫染色的增强。IL-1刺激诱导细胞溶质Ca2+剂量依赖性增加并激活蛋白酪氨酸磷酸化。细胞内Ca2+螯合可阻止IL-1依赖性的Cx43上调,但蛋白酪氨酸激酶抑制剂则不能,这表明细胞溶质Ca2+在调节Cx43表达中起关键作用。IL-1是促进软骨分解代谢最有效的细胞因子之一;其对细胞间通讯的调节代表了促炎介质调节软骨细胞活性的一种新机制。