Nishida Yoshihiro, Knudson Cheryl B, Knudson Warren
Department of Biochemistry, Rush Medical College, Rush University Medical Center, 1653 W. Congress Parkway, Chicago, IL 60612, USA.
Osteoarthritis Cartilage. 2004 May;12(5):374-82. doi: 10.1016/j.joca.2004.01.008.
To examine the capacity of recombinant osteogenic protein-1 (OP-1) to inhibit the cartilage extracellular matrix damage that follows treatment with hyaluronan hexasaccharides (HA6).
The effects of OP-1 were examined on isolated human chondrocytes grown in alginate beads as well as articular cartilage slices treated with hyaluronan hexasaccharides. Changes in the relative expression of messenger RNA for hyaluronan synthase- 2, aggrecan and CD44 were determined by competitive quantitative reverse transcriptase-polymerase chain reaction. Cartilage proteoglycan biosynthesis was examined by a (35)S-sulfate incorporation assay. Cell-associated matrix of human chondrocytes was visualized by the use of particle exclusion assay, and alcian blue staining. Cartilage slices were examined for accumulation of proteoglycan by Safranin-O, and hyaluronan by a specific biotinylated probe.
Combined OP-1 and HA6 treatment resulted in enhanced expression of mRNA for aggrecan and HAS-2, compared to the treatment with HA6 only. This increased expression of aggrecan mRNA was paralleled by an increased synthesis of cartilage proteoglycan especially retained in the cell-associated matrix. Co-treatment with OP-1 inhibited the HA6-induced depletion of cell-associated matrices as well as HA6-induced depletion of hyaluronan and proteoglycan within cartilage tissue slices.
These results demonstrate that OP-1 can abrogate the catabolic events associated with a HA6-induced matrix depletion model of osteoarthritis. The mRNA levels of two major cartilage extracellular matrix components, aggrecan and hyaluronan synthase-2 are enhanced above values obtained by either OP-1 or HA6 treatments alone.
研究重组骨生成蛋白-1（OP-1）抑制透明质酸六糖（HA6）处理后软骨细胞外基质损伤的能力。
检测OP-1对在藻酸盐珠中培养的分离人软骨细胞以及用透明质酸六糖处理的关节软骨切片的作用。通过竞争性定量逆转录聚合酶链反应测定透明质酸合酶-2、聚集蛋白聚糖和CD44信使核糖核酸的相对表达变化。通过（35）S-硫酸盐掺入试验检测软骨蛋白聚糖的生物合成。使用颗粒排除试验和阿尔新蓝染色观察人软骨细胞的细胞相关基质。用番红O检测软骨切片中蛋白聚糖的积累，并用特异性生物素化探针检测透明质酸。
与仅用HA6处理相比，OP-1和HA6联合处理导致聚集蛋白聚糖和HAS-2信使核糖核酸表达增强。聚集蛋白聚糖信使核糖核酸表达的增加与软骨蛋白聚糖合成的增加平行，尤其是保留在细胞相关基质中的合成增加。OP-1共同处理抑制了HA6诱导的细胞相关基质的消耗以及HA6诱导的软骨组织切片中透明质酸和蛋白聚糖的消耗。
这些结果表明，OP-1可以消除与HA6诱导的骨关节炎基质消耗模型相关的分解代谢事件。两种主要软骨细胞外基质成分，聚集蛋白聚糖和透明质酸合酶-2的信使核糖核酸水平比单独用OP-1或HA6处理所获得的值有所提高。
