大鼠髋脱位在退变过程中软骨细胞增殖与凋亡的变化及意义  

Proliferation and apoptosis of chondrocytes in dislocational hip of neonatal rats in the process of degeneration

作　　者：王彭[1] 宁波[1] 裴新红[1] 马瑞雪[1] 

机构地区：[1]复旦大学附属儿科医院骨科,上海211102

出　　处：《中华小儿外科杂志》2013年第12期938-943,共6页Chinese Journal of Pediatric Surgery

基　　金：基金项目：国家自然科学基金（3{1973138）;上海市科委自然基金（9411963200）

摘　　要：目的探讨新生大鼠髋脱位在自然演变过程中关节软骨细胞的增殖能力和凋亡的变化及意义。方法将新生大鼠按数字随机法随机分成实验组和对照组，实验组模拟襁褓位固定双下肢伸髋伸膝位持续10天。解除外固定后每鼠龄2、4、8周处死并离断髋关节，一侧用于大体形态测量与番红。固绿染色评估；另一侧用于体外原代细胞培养，流式分析增殖指数与凋亡率；CCK8计算增殖速率。采用t检验进行统计分析，P〈0．05为差异有统计学意义。结果①大体形态：实验组大鼠去除外固定后未获得复位，股骨头一直脱出于髋臼外上方，对照组髋关节臼深指数随着生长发育逐渐增大，而实验组在4周时明显增大后至8周时却又明显变小，并且与不同时段相比明显小于对照组，差异有统计学意义（P〈0．01）。对照组股骨头形态指数随生长发育无明显变化，而实验组逐渐减小，并明显小于对照组。②组织形态学：番红一固绿染色可见对照组随着生长发育，关节软骨逐渐变薄，且层次清晰；实验组在4周软骨形态失常，且浅层基质淡染，细胞表现出排列紊乱，局部出现细胞聚集成簇；8周浅层软骨出现大面积的失染，浅层出现软骨细胞丢失，伴随裂隙和溃疡，移行层细胞却增多；Mankin评分显示在4、8周阶段实验组的评分显著高于对照组（P〈0．01）。③细胞增殖能力分析：流式分析结果显示不同时段实验组的增殖指数均明显高于对照组；CCK-8也显示在不同时段实验组的增殖速率高于对照组，差异有统计学意义（P〈0．01）。④凋亡检测：流式分析显示在鼠龄2、8周阶段实验组凋亡率明显高于对照组（P〈0．01）；而在4周阶段却无显著差异。结论新生大鼠髋脱位关节软骨细胞表现出较强的增殖活性与异常增高的凋亡对早期发生退行性改变有着重要意义。Objective To explore the proliferation and apoptosis of chondrocytes during the de- genration process resulted from dislocation of the hip in neonatal rat model. Methods Neonatal Wistar rats were randomly divided into two groups （experimental and control group）. Developmental disloca- tion of the hip （DDH） model was mimicked by swaddling position, which was to immobilize the bilat eral hips and legs at extension with medical tape for ten days. The rats were sacrificeal at 2, 4 and 8 weeks of age and hips were isolated for gross measurement and sanfranin （）-fast green staining. Chon- drocytes were isolated for proliferation index （PI） and apoptosis ratio （AR） by flow cytometry （FCM） and CCK-8 test. Student＇s-t test was applied for statistical analysis. Results The gross sample showed the abnormal relationship of the hips, proving the success of model. The anatomy didn＇t get reduction with development. Acetabula depth ratio （ADR） of control group increased along with de- velopment, however, ADR in DDH group showed an obvious increase at 4-weeks with a deep decrease at 8 weeks and were smaller than those in the control group at each stage. Femoral head ratio （FHR） in control group didn＇t show significant changes along with growing up; however, FHR in DDH group decreased along with development and was smaller than those in the control group. The sanfranin O- fast green staining revealed that the cartilage in control group became thinner and showed distinct lay ers along with development, whereas the shape of cartilage in DDH group deformed from 4 weeks afterbirth and extracellular matrix （ECM） became less stained, chondrocytes disordered and clusters pres- ented. Mankin score showed significant differenee at 4 and 8 weeks after birth （P〈2（）. Ill ）. PI of DDH group showed higher than those in tile conlrol group at each stage, as well as proliferative ratio by CCK 8 lest. AR in I）DH group was higher than that in control group al 2 and 8 weeks, while no sig nifieance

关 键 词：髋脱位 软骨 关节 大鼠 

分 类 号：R726.5[医药卫生—儿科]