CAJ | 학술논문

目的：观察兔退变椎间盘纤维环和髓核组织中蛋白多糖（PG）、基质金属蛋白酶-3（MMP-3）、基质金属蛋白酶组织抑制因子-2（TIMP-2）的表达，并探讨其意义。方法新西兰大白兔42只，直立位下建立椎间盘退变模型，均分为实验组和对照组。两组分别于建模后4、8、12周各取7只动物，取 L4～5椎间盘组织采用高碘酸-希夫（PAS）染色检测髓核与纤维环组织中的 PG，以灰度值表示其相对表达量；免疫组化 SP法检测椎间盘纤维环和髓核组织中的 MMP-3及 TIMP-2，以平均光密度值（MOD）表示其相对表达量。结果实验组、对照组建模4、8、12周髓核组织中 PG灰度值均高于纤维环组织（P＜0．05）；两组建模12周时纤维环、髓核PG灰度值均高于8周时，且建模8周时髓核 PG灰度值高于4周时（P＜0．05）。随着建模时间的延长，实验组中MMP-3和TIMP-2的相对表达量均增加明显。髓核中，对照组及实验组同一时间点 MMP-3的平均 MOD值明显高于 TIMP-2。实验组纤维环中TIMP-2较 MMP-3表达变化明显。结论椎间盘退变过程中髓核中PG的含量变化早于纤维环，通过对 MMP-3/TIMP-2在椎间盘退变中所起作用的研究结合髓核 PG的变化可能会对临床诊断、基因治疗和组织工程技术的应用有一定帮助。而且对 MMPs和TIMPs的表达水平的调节来控制椎间盘基质代谢的平衡，可能会成为治疗椎间盘退变的新方向。
목적：관찰토퇴변추간반섬유배화수핵조직중단백다당（PG）、기질금속단백매-3（MMP-3）、기질금속단백매조직억제인자-2（TIMP-2）적표체，병탐토기의의。방법신서란대백토42지，직립위하건립추간반퇴변모형，균분위실험조화대조조。량조분별우건모후4、8、12주각취7지동물，취 L4～5추간반조직채용고전산-희부（PAS）염색검측수핵여섬유배조직중적 PG，이회도치표시기상대표체량；면역조화 SP법검측추간반섬유배화수핵조직중적 MMP-3급 TIMP-2，이평균광밀도치（MOD）표시기상대표체량。결과실험조、대조조건모4、8、12주수핵조직중 PG회도치균고우섬유배조직（P＜0．05）；량조건모12주시섬유배、수핵PG회도치균고우8주시，차건모8주시수핵 PG회도치고우4주시（P＜0．05）。수착건모시간적연장，실험조중MMP-3화TIMP-2적상대표체량균증가명현。수핵중，대조조급실험조동일시간점 MMP-3적평균 MOD치명현고우 TIMP-2。실험조섬유배중TIMP-2교 MMP-3표체변화명현。결론추간반퇴변과정중수핵중PG적함량변화조우섬유배，통과대 MMP-3/TIMP-2재추간반퇴변중소기작용적연구결합수핵 PG적변화가능회대림상진단、기인치료화조직공정기술적응용유일정방조。이차대 MMPs화TIMPs적표체수평적조절래공제추간반기질대사적평형，가능회성위치료추간반퇴변적신방향。
Objective To observe the expression of proteoglycan (PG),MMP-3,TIMP-2 in the rabbit degenerative interverte-bral disc fibrous ring and nucleus pulposus tissues and to investigate their significance.Methods 42 New Zealand white rabbits were used for establishing the intervertebral disc degeneration model under the upright position and equally divided into the experi-mental group and the control group.The L4-5 intervertebral disc tissues were taken in 7 rabbits per group at 4,8,12 weeks after modeling.The periodate Schiff (PAS)staining was adopted to detect PG in the nucleus pulposus and fibrous ring tissues with the grayscale value for expressing the relative expression quantity;the immunohistochemical SP method was adopted to detect MMPs-3 and TIMP-2 in the disc fibrous ring and nucleus pulposus tissues,with the mean optical density(MOD)value for expressing the relative expression quantity.Results The PG gray values in the nucleus pulposus tissue after 4,8,12 weeks in the experimental group and the control group model were higher than those in the fiber ring tissue (P<0.05);the PG gray values in the nucleus pulposus and fiber ring tissue at 1 2 weeks were higher than those at 8 weeks,and at 8 weeks of modeling,the PG gray value in the nucleus pulposus was higher than that at 4 weeks (P<0.05).As the time extension of modeling,the relative expression amounts of MMP-3 and TIMP-2 in the experimental group were significantly increased.The average MOD value of MMP-3 in the nucleus pul-posus at the same time point in the control group and the experimental group was obviously higher than that of TIMP-2.The ex-pression of TIMP-2 in the fibrous ring in the experimental group had more change than the expression of MMP-3.Conclusion Dur-ing the intervertebral disc degeneration process,the change of PG content in the nucleus pulposus is earlier than that of the fibrous ring.The study of the MMP3/TIMP-2 role in the intervertebral disc degeneration in combination with the changes of nucleus pulpo-sus PG may conduce to clinical diagnosis,gene therapy and the application of tissue engineering technique.Moreover regulating the expression level of MMPs and TIMPs for controlling the balance of intervertebral disc matrix metabolism could become the new di-rection for treating the intervertebral disc degeneration.