羟基喜树碱对兔膝关节粘连中成纤维细胞的影响研究

Hydroxycamptothecin reduces intraarticular scar adhesion in a rabbit model of knee synarthrophysis

目的 探讨局部应用羟基喜树碱对兔膝关节粘连中成纤维细胞的影响. 方法 将24只新西兰大白兔随机分成2组（n=12）：去除右侧股骨髁两侧约10 mm＆#215;10mm的骨皮质,暴露松质骨.然后将浸透有0.1 mg/mL羟基喜树碱（实验组）或生理盐水（对照组）的棉片局部应用于手术区域.4周后通过组织学观察来评价羟基喜树碱对膝关节粘连瘢痕组织中胶原及成纤维细胞的影响.从兔膝关节瘢痕组织中分离、培养成纤维细胞,将所得成纤维细胞分为2组（n=6）：0.1 mg/L羟基喜树碱组（应用0.1 mg/L羟基喜树碱处理）和空白组（不用药物,仅进行换液处理）,24 h后应用细胞流式仪检测羟基喜树碱对成纤维细胞凋亡率的影响. 结果 实验组兔膝关节粘连瘢痕组织中胶原含量明显低于对照组,成纤维细胞数量为（15.7±4.6）个,显著低于对照组[（30.7±5.2）个],差异有统计学意义（t =5.274,P＜0.001）.0.1 mg/L羟基喜树碱组成纤维细胞的凋亡率为18.2％±0.7％,明显高于空白组（8.2％±0.4％）,差异有统计学意义（t=30.442,P＜0.001）. 结论 在兔膝关节粘连模型中应用羟基喜树碱可以通过抑制成纤维细胞的增殖、诱导成纤维细胞的凋亡来减少膝关节粘连.

Objective To study the effect of topical application of hydroxycamptothecin on the fibroblasts in a rabbit model of knee synarthrophysis.Methods Twenty-fiour mature male New Zealand white rabbits,weighing 2.0 to 2.5 kg,were used for this experiment.They were randomly and equally divided into 2 groups：a 0.1 mg/mL hydroxycamptothecin group and a control （saline） group.Approximately 10 mm ＆#215; 10 mm of the cortical bone was removed in each rabbit from both sides of the right femoral condyle to expose the underneath cancellous bone.Hydroxycamptothecin （0.1 mg/mL） or saline was applied to the cancellous bone for 10 min.After 4 weeks,all animals were sacrificed.The collagen density and fibroblasts of the intraarticular adhesion were assessed by histological evaluation.The fibroblasts were isolated from the intraarticular adhesion scar and cultured.The fibroblasts harvested were divided into 2 groups （n =6） which were subjected to treatment of 0.1 mg/L or 0 mg/L hydroxycamptothecin.Cell apoptosis was detected by flow cytometry after 24 hours.Results The collagen density of the intraarticular tissue in the experimental group was obviously lower than in the control group.The fibroblasts in the 0.1 mg/mL hydroxycamptothecin group were 15.7 ±4.6,significantly fewer than those（30.7 ± 5.2） in the control groups（t =5.274,P 〈 0.001）.The apoptosis rate of fibroblasts after 24 hours＇ culture in the 0.1 mg/L hydroxycamptothecin group （18.2% ±0.7%） was significantly higher than that （8.2% ±0.4%） in the control groups （t=30.442,P 〈 0.001）.Conclusion Topical application of hydroxycamptothecin may reduce intraarticular scar adhesion by inhibiting proliferation of fibroblasts and inducing apoptosis of fibroblasts in a rabbit model of knee synarthrophysis.