基质金属蛋白酶9在大鼠跟腱异位骨化模型中的动态变化研究  

DYNAMIC CHANGES OF MATRIX METALLOPROTEINASE 9 IN HETEROTOPIC OSSIFICATION OF RAT MODEL

作　　者：石伟哲[1,2] 肖海军[1,2] 薛锋[1,2] 吴江[1,2] 

机构地区：[1]南方医科大学第三临床医学院 [2]南方医科大学附属奉贤医院骨科,上海201400

出　　处：《中国修复重建外科杂志》2014年第9期1133-1138,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：上海市卫生局科研课题资助项目(20134204)~~

摘　　要：目的通过观测创伤早期大鼠异位骨化模型中基质金属蛋白酶9（matrixmetalloproteinase9，MMP-9）表达变化，探讨MMP-9对预测异位骨化发生的价值。方法取4～5周龄雄性SD大鼠132只，体重（135．0±6．5）g，随机分为两组（n=66）。实验组通过钳夹并切断跟腱制备异位骨化模型，对照组仅暴露跟腱。术后观察两组大鼠一般情况，2、3、4、5、6、7、8d两组各取8只大鼠，取跟腱组织行大体、组织学及免疫组织化学染色观察，取血清及跟腱组织分别行ELISA及RT-PCR检测MMP-9蛋白含量及基因表达。通过两组剩余10只大鼠术后5、10周x线片及术后10周组织学观察结果，诊断异位骨化形成情况。结果所有大鼠均存活至实验完成。各时间点对照组大鼠跟腱无明显变化，呈正常跟腱结构；实验组随时间延长，跟腱断端硬度逐渐增加，出现大量不规则结缔组织及软骨细胞，MMP-9免疫组织化学染色呈深染阳性。各时间点实验组MMP-9蛋白含量及基因相对表达量均显著高于对照组（P〈0．05）；实验组MMP-9蛋白含量及基因相对表达量均总体呈上升趋势（P〈0．05）。根据x线片及组织学观察结果，术后10周实验组大鼠跟腱组织均发生异位骨化，对照组均无异位骨化发生。结论在大鼠跟腱异位骨化早期，MMP-9表达明显升高，提示其对预测异位骨化发生有参考意义。Objective To explore the value of matrix metalloproteinase 9 （MMP-9） in predicting the occurrence of heterotopic ossification by observing the expression of MMP-9 in heterotopic ossification of the early trauma rat model. Methods A total of 132 male Sprague Dawley rats, aged 4-5 weeks, weighing （135.0 ± 6.5） g, were randomly divided into experimental group and control group （n=66）. In experimental group, the Achilles tendon was cut offand clamped to prepare heterotopic ossification model; in control group, only Achilles tendon was exposed by making a incision. The general condition of the rats was observed after operation; at 2, 3, 4, 5, 6, 7, and 8 days after operation, the Achilles tendon tissue was harvested for gross observation, histological observation, and immunohistochemical staining observation; the serum and Achilles tendon tissue were harvested to detect the expressions of MMP-9 protein and mRNA by ELISA and RT-PCR. The X-ray films at 5 and 10 weeks and histological examination at 10 weeks after operation were used to observe heterotopic ossification. Results All rats survived to the end of the experiment. The Achilles tendon had no significant change in control group at each time point, showing normal tendon structure. In experimental group, the hardness of Achilles tendon tissue gradually increased with the time; there were a large number of irregular connective tissue and cartilage cells; and immunohistochemical staining for MMP-9 was positive results. The MMP-9 protein and mRNA expression levels of experimental group were significantly higher than those of the control group at each time point （P 〈 0.05）. MMP-9 protein and mRNA expression levels of experimental group showed an increasing tendency （P 〈 0.05）. According to the results of X-ray films and histological observation, heterotopic ossification occurred at 10 weeks after operation in experimental group, but no heterotopic ossification was observed in control group. Conclusion In early heterotopic ossification of rat A

关 键 词：基质金属蛋白酶9 异位骨化 细胞外基质 大鼠 

分 类 号：R-332[医药卫生] R641