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[大鼠模型异位骨化中基质金属蛋白酶9的动态变化]

[Dynamic changes of matrix metalloproteinase 9 in heterotopic ossification of rat model].

作者信息

Shi Weizhe, Xiao Haijun, Xue Feng, Wu Jiang

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2014 Sep;28(9):1133-8.

Abstract

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 off and 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 Achilles tendon, the expression of MMP-9 increases significantly, indicating that it has reference significance in predicting heterotopic ossification.

摘要

目的

通过观察基质金属蛋白酶9(MMP - 9)在早期创伤大鼠模型异位骨化中的表达情况,探讨其在预测异位骨化发生中的价值。

方法

选取132只4 - 5周龄、体重(135.0±6.5)g的雄性Sprague Dawley大鼠,随机分为实验组和对照组(n = 66)。实验组切断并钳夹跟腱制备异位骨化模型;对照组仅切开暴露跟腱。术后观察大鼠一般情况;术后2、3、4、5、6、7、8天,取跟腱组织进行大体观察、组织学观察及免疫组织化学染色观察;采集血清及跟腱组织,采用酶联免疫吸附测定法(ELISA)和逆转录聚合酶链反应(RT - PCR)检测MMP - 9蛋白及mRNA表达。术后5周和10周拍摄X线片,术后10周进行组织学检查以观察异位骨化情况。

结果

所有大鼠均存活至实验结束。对照组各时间点跟腱无明显变化,腱结构正常。实验组跟腱组织硬度随时间逐渐增加;有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阳性结果。实验组各时间点MMP - 9蛋白及mRNA表达水平均显著高于对照组(P < 0.05)。实验组MMP - 9蛋白及mRNA表达水平呈上升趋势(P < 0.05)。根据X线片及组织学观察结果,实验组术后10周发生异位骨化,而对照组未观察到异位骨化。

结论

在大鼠跟腱早期异位骨化中,MMP - 9表达显著增加,表明其在预测异位骨化方面具有参考意义。

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