探讨基质金属蛋白酶3(matrixmetalloproteinase3,MMP-3)在强直性脊柱炎(ankylosingspondylitis,AS)棘上韧带中的表达及在韧带退变中的作用。方法免疫组化检测MMP-3、I型胶原在韧带中的表达,酶联免疫吸附试验(enzymelinkedimmunosorbentassay,ELISA)检测韧带中蛋白聚糖(proteoglycan)的含量,扫描电镜及透射电镜对棘上韧带进行形态学观察。Image—ProPlus(IPP)量化分析胶原原纤维分布密度及胶原阳性染色面积百分比。结果MMP-3在研究组韧带血管平滑肌细胞及内皮细胞中高表达,对照组未见表达;蛋白聚糖含量对照组为(19.462±2.174)μg/mg,研究组为(7.477±2.512)μg/mg;电镜下对照组胶原纤维排列规律,研究组胶原原纤维裸露明显,大量基质降解;对照组胶原原纤维分布密度为(218.02±10.39)/μm2,研究组为(122.47±22.42)/μm。对照组I型胶原的阳性染色面积百分比为(60.23±3.41)%,研究组为(43.37±5.96)%,2组差异有统计学意义(P〈0.05)。结论活动期AS患者棘上韧带血管中MMP-3高表达,可能通过促进血管生长、降解蛋白聚糖、破坏韧带正常的微观结构,从而参与韧带退变骨化的病理过程。
Objective To study the expression of matrix metalloproteinase 3 (MMP-3) and its effect on supraspinal ligament degeneration due to ankylosing spondylitis (AS). Methods Expression of MMP-3 and type I collagen in ligaments was detected by immunohistochemistry. Proteoglycan level in ligaments was measured by ELISA. Supraspinal ligament morphology was observed under scanning electron microscope and transmission electron microscope, respectively. Collagen fibril distribution density and positively-stained collagen area were analyzed by Image-Pro Plus (IPP). Results MMP-3 was highly expressed in vascular smooth muscle cells and endothelial cells in AS group but not in control group. The proteoglyean level was significantly higher in control group than in AS group ( 19. 462 ± 2. 174 vs 7. 477 ± 2. 512 μg/mg). The collagen fibrils were regularly arranged in control group and apparently exposed with massive degradation of matrix in AS group. The distribution density of collagen fibrils was 218.02 ± 10.39/pjn2 and 122.47 ± 22.42/μm2 in control group and AS group, respectively. The proportion of positively-stained collagen area was significantly higher in control group than in AS group [ (60.23 ± 3.41 ) % vs (43.37±5.96) %, P 〈 0.05 ]. Conclusion MMP-3 is highly expressed in supraspinal ligament blood vessels of active AS patients by promoting the growth of blood vessels, activating the degradation of proteoglycan, and destroying the normal ultra-structure of ligaments, and is thus involved in the degenerative ossification process of ligaments.