摘要
目的：以Ｒｈｏｄａｍｉｎｅ标记的核心蛋白作为荧光探针，观察腰椎间盘中的蛋白多糖聚合体的分布。方法：将小牛的肩胛软骨溶于含蛋白酶抑制剂的４．０Ｍ盐酸胍中，通过分解状态下的高速离心、透析、聚合状态下的高速离心提取出蛋白多糖聚合体。然后经分解状态下的超速离心、酶解、层析及与荧光素耦合等方法制备出一种带有荧光素的核心蛋白，作为透明质酸的特异性荧光探针来观察椎间盘中蛋白多糖聚合体的分布。结果：胎儿、正常成年人及椎间盘突出症病人的椎间盘中蛋白多糖聚合体的形态学分布是不同的。另外，在细胞表面及胶原纤维间的特殊部位也有蛋白多糖聚合体的分布。结论：蛋白多糖聚合体在椎间盘中的分布随年龄增加及退变而减少。
bjective:To observe the changes of morphological distribution of human lumbar disc proteoglycan aggregates with fluoresent probe. Methods: Calfscapular cartilage was extracted with 40M guanidine hydrochloride which contained protease inhibitors. Proteoglycan aggregates were extracted by highspeed centrifugation underdissociative conditions,dialyzing and highspeed centrifugation underassociative conditions. Then,proteoglycan monomer and core protein with fluorescent,a kind of fluorescent probe for proteoglycan aggregate were prepared by ultracentrifugation under dissociative conditions,enzymolysis,chromatographing and conjugating with fluorescein. With the fluorescent probe,distribution of proteoglycan aggregates in human lumbar intervertebral disc were observed. Results: It was found that proteoglycan aggregates in both annulus fibrosis and nucleus pulposus distributed on the cellular surface and in the extracellular matrix. The distribution of proteoglycan aggregatesis different between fetal lumbar disc and that of the adult. In the latter,proteoglycan aggregates distributed in parallel with the collagen fibrils. In discs from patients suffered from lumbar disc herniation,few of proteoglycan aggregates were observed. Conclusion: Proteoglycan aggregates of human lumbar disc decreased with age and degeneration.
出处
《中华骨科杂志》
CAS
CSCD
北大核心
1998年第8期460-462,I001,共4页
Chinese Journal of Orthopaedics
关键词
腰椎
椎间盘
蛋白多糖聚合体
荧光探针
uman Lumbar vertebrae Intervertebral disc Proteoglycans
