一氧化氮合酶抑制剂对软骨修复组织胶原表达的影响

Influence of collagenous expression by iNOS inhibitor in repaired articular cartilages

目的探讨应用苦味酸-天狼猩红染色观察诱导型一氧化氮合酶(iNOS)抑制剂对关节软骨修复组织胶原表达的影响及胶原定量表达的新方法。方法24只新西兰大白兔双侧股骨髁间关节面造成全层软骨缺损,随机均分为对照组、骨形态发生蛋白(BMP)组和iNOS抑制剂S-甲基异硫脲(SMT)组。术后16周及1年各组分别处死4只动物。应用苦味酸-天狼猩红染色检测胶原的分布情况,图像分析技术定量胶原的表达及进行修复组织软骨厚度的测量。结果图像分析结果经GLM-GeneralFactorialANOVA作均数的显著性检验,显示术后16周SMT组Ⅰ型胶原表达量为61.8%,明显少于BMP组的83.6%,和对照组的89.4%相比,差异有统计学意义(F=23.562);术后1年SMT组Ⅰ型胶原表达量为65.9%,明显少于BMP组88.5%和对照组94.6%,差异有统计学意义(F=36.747)。术后16周SMT组Ⅱ型胶原表达量为36.5%,明显多于BMP组12.6%和对照组8.2%,差异有统计学意义(F=68.326);术后1年SMT组Ⅱ型胶原30.7%的表达量多于BMP组的10.8%和对照组的4.5%,差异有统计学意义(F=83.876)。术后16周及1年后SMT组软骨厚度(2.08±0.6,1.85±0.56),与BMP组(1.56±0.38,0.67±0.31)和对照组(0.64±0.43,0.24±0.10)相比,差异有统计学意义(F值分别为57.836,45.087)。结论iNOS抑制剂SMT的应用能明显增加Ⅱ型胶原表达和软骨厚度,对于维持软骨表型有积极意义。而苦味酸-天狼猩红染色是一种观察软骨修复组织中胶原表达的理想方法。

Objective To observed the effect of inducible nitric oxide synthase inhibitor-SMT on collagenous expression in repaired articular cartilages using Sirius-Red stain, and explore a semi-quantitative way of collagenous expression. Methods Twenty-four adult New Zealand White rabbits which had full-thickness defects of cartilage were created in the bilateral trochlear groove were randomly divided into the control, BMP group and SMT group. Four rabbits in each group were executed with anesthesia respectively at sixteen-weeks and one-year after operation. All tissue sections were was stained with Sirius-Red against collagen type-Ⅰ expression and type-Ⅱexpression which were analyzed by Quantiment 500 image analysis system. Cartilage thickness was measured by Quantiment 500 too. Results Sixteen-week and one-year after operation, collagen type-Ⅰexpression of repaired articular cartilage in SMT group（61.8% ,65.9% ） was less than that in BMP group （83.6% ,88.5% ） and the control group （89.4% ,94. 6% ）, and collagen type-Ⅱ expression （36. 5% ,30. 7% ） was much more than in BMP group（ 12. 6%, 10. 8% ） and the control group （8. 2% ,4. 5% ）. Cartilage thickness of SMT group （2.08±0. 6,1.85±0. 56 ） was significant higher than that of BMP group（1.56±0.38,0.67±0.31） and the control group（0.64±0.43,0.24±0. 10）. Conclusions NOS inhibitor SMT could increase collagen type-Ⅱ expression and decrease collagen type-Ⅰ expression, as well as make phenotype well. Sirius-Red stain is a good way to observe collagenous expressions in repaired articular cartilage.