椎间盘体外培养方法及其初步应用的研究

Study on a Culture Method of Intervertebral Disc In Vitro and Its Initial Application

[目的 ]探讨一种新的椎间盘藻酸盐体外培养方法。同时将这种培养方法初步应用于椎间盘退变的研究 ,以进一步了解退变的表现和可能的机制。[方法 ]取大鼠椎间盘分置于藻酸盐和对照培养基进行培养 ,应用不同剂量的萘啶酸退变剂对藻酸盐培养方法培养的椎间盘进行处理 ,分别测定上述两个实验过程中相关生化指标的改变。[结果 ]藻酸盐方法培养的椎间盘其胶原含量、含水率、碱性磷酸酶 (alkalinephosphatase ,AKP)活性随培养时间延长无明显变化 (P>0 0 5 ) ;而在对照组 ,以上指标出现了明显降低 (P <0 0 1)。随退变剂浓度的升高 ,羟脯氨酸 (hydroxyproline ,HYP)、一氧化氮 (nitricoxide ,NO)含量、谷胱甘肽过氧化物酶 (glutathioneperoxidase ,GSH Px)活性升高。[结论 ]藻酸盐培养方法有利于维持椎间盘正常的代谢和功能。HYP的增加可能是椎间盘退变的表现之一 ,而NO和GSH Px在椎间盘退变中可能起到某些重要作用。

Objective] To study a new culture method of intervertebral disc with alginate in vitro and using the method to study initially the manifestation and probable mechanism of disc degeneration. [Method]Lumbar discs separated from Wistar rats were cultured with and without alginate;secondly,nalidixic acid known as a definite degeneration causing agent was added to the disc cultured with alginate,whereby some biochemical changes were determined successively. [Results] With prolongation of culture time,there were no significant changes in contents of collagen,water proportion and the activities of AKP in alginate group ( P>0 05 ),but they decreased dramatically in the control group ( P<0 01 ).By rise of nalidixic acid concentration,the contents of HYP and NO,the activities of GSH Px were found increased significantly. [Conclusion] Alginate culture method is better than general method to promote the retention of normal structure and metabolic activities of the intervertebral disc.The increase of the content of HYP is probably a main degenerative manifestation of intervertebral disc,NO and GSH Px may play important role in the processes.