Carney S L, Billingham M E, Caterson B, Ratcliffe A, Bayliss M T, Hardingham T E, Muir H
Biochemistry Division, Kennedy Institute of Rheumatology, Hammersmith, London, UK.
Matrix. 1992 Apr;12(2):137-47. doi: 10.1016/s0934-8832(11)80055-7.
The metabolism of newly-synthesised and total ("resident") proteoglycans was examined in control and osteoarthritic cartilage explants obtained from an experimental model (Pond and Nuki, 1973) of canine osteoarthritis. The following findings were obtained: (i) Non-labelled proteoglycans extracted from normal cartilage with 4 M guanidine HCl showed two bands visualised by staining with toluidine blue. The electrophoretic mobilities of proteoglycans from osteoarthritic cartilage were unchanged but the relative abundance of the slower migrating band increased with time after surgery. (ii) There were qualitative differences in the proteoglycan breakdown products released into the medium of explant cultures of osteoarthritic compared with control cartilage. This was apparent for both labelled and total unlabelled proteoglycans. (iii) There were similarities in the electrophoretic mobilities of the major labelled and non-labelled proteoglycan breakdown products suggesting that total ("resident") proteoglycans and newly-formed proteoglycans were degraded by similar mechanisms. There were however some differences in the labelled and non-labelled proteoglycans, suggesting that the mechanisms of breakdown were not identical. (iv) Immunoblotting techniques showed differences in the distribution of various glycosaminoglycans in proteoglycan breakdown products from control compared with osteoarthritic cartilage explant cultures. (v) Monoclonal antibodies 7-D-4 and 3-B-3 (which recognise unusual native chondroitin sulphate epitopes) showed greatly increased expression on proteoglycans from osteoarthritic cartilage compared with controls.
在从犬骨关节炎实验模型(Pond和Nuki,1973)获取的对照和骨关节炎软骨外植体中,研究了新合成的和总的(“驻留”)蛋白聚糖的代谢情况。得到了以下研究结果:(i)用4M盐酸胍从正常软骨中提取的未标记蛋白聚糖,经甲苯胺蓝染色后显示出两条带。骨关节炎软骨中蛋白聚糖的电泳迁移率没有变化,但手术后随着时间的推移,迁移较慢的条带的相对丰度增加。(ii)与对照软骨相比,释放到骨关节炎外植体培养物培养基中的蛋白聚糖降解产物存在质的差异。这在标记的和总的未标记蛋白聚糖中都很明显。(iii)主要的标记和未标记蛋白聚糖降解产物的电泳迁移率存在相似性,这表明总的(“驻留”)蛋白聚糖和新形成的蛋白聚糖通过相似的机制降解。然而,标记的和未标记的蛋白聚糖之间存在一些差异,这表明降解机制并不完全相同。(iv)免疫印迹技术显示,与骨关节炎软骨外植体培养物相比,对照中蛋白聚糖降解产物中各种糖胺聚糖的分布存在差异。(v)单克隆抗体7-D-4和3-B-3(识别异常的天然硫酸软骨素表位)显示,与对照相比,骨关节炎软骨中的蛋白聚糖表达大幅增加。