摘要
背景:关节软骨细胞凋亡的异常表达会引起软骨退变,最终发展为骨性关节炎。目的:观察发育性髋关节脱位模型经过髋关节复位法处理后细胞凋亡相关因子的表达情况。方法:随机选取28日龄的新西兰大白兔60只,雌雄不限,在兔右后肢屈髋伸膝位时用管型石膏固定8周制作髋关节脱位动物模型,固定的右后肢作为实验组,同只兔的左后肢不做任何处理作为对照组。8周后根据X射线片、髋臼指数、股骨头脱位程度判断造模情况。选取造模成功的48只兔,根据兔右后肢不同的处理方式,将实验组再分为A组(固定8周)、B组(固定10周)、C组(固定8周后解除固定复位2周),每组16只。实验结束后苏木精-伊红染色观察髋臼软骨软骨细胞形态;免疫组织化学法检测软骨细胞中Caspase-3、Bcl-2的表达;TUNEL法检测髋臼软骨中凋亡率的表达水平;并进行相关性分析。结果与结论:(1)A、B、C实验组髋臼指数均较其对照组明显增大(P <0.05);(2)苏木精-伊红染色后对照组软骨细胞形态规整,A、B实验组出现空泡细胞,部分结构消失,C实验组髋臼和股骨头关节软骨细胞结构较完整,空泡现象较少;(3)TUNEL结果示C实验组的凋亡率低于B实验组,但高于其对照组(P <0.05),说明经过复位法处理后凋亡率有所降低;(4)相关性分析示实验组细胞凋亡率与Caspase-3表达水平呈正相关,与Bcl-2表达水平呈负相关;(5)结果说明,复位组较固定8周、固定10周Bcl-2高表达,凋亡率较固定10周低表达,说明复位处理后软骨细胞凋亡速度放缓,同时凋亡效应蛋白Caspase-3较固定10周低表达,凋亡程度有所降低。复位法可降低髋关节脱位中髋臼软骨细胞凋亡程度的表达水平,延缓软骨进一步退变的进程,从而减缓患者的病情恶化的病程。
BACKGROUND: Abnormal increase in the number of apoptotic articular chondrocytes can induce cartilage degeneration, further deteriorate and develop into osteoarthritis.OBJECTIVE: To observe the expression of apoptosis-related factors in the treatment of developmental dysplasia of the hip by reduction.METHODS: Sixty 28-day-old New Zealand white rabbits(either sex) were randomly selected, then the right hind limbs were fixed with tubular gypsum in flexion of hip and extension of knee for 8 weeks to establish an animal model of developmental dysplasia of the hip, and the left hind limbs served as controls. Subsequently, the successful models were judged by the X-ray film, acetabular index, and dislocation of the femoral. Forty-eight model rabbits were equally randomized into group A(8 weeks of fixation), group B(10 weeks of fixation), group C(embolia at 2 weeks after 8 weeks of fixation). The morphology of chondrocytes at the acetabular cartilage was observed after hematoxylin-eosin staining. The expression levels of Caspase-3 and Bcl-2 in chondrocytes were detected by immunohistochemistry. The apoptosis rate in the acetabular cartilage was detected by TUNEL method, and the correlation was analyzed.RESULTS AND CONCLUSION:(1) The acetabular index in the groups A, B and C was significantly higher than that in the control group(P <0.05).(2) The morphology of chondrocytes in control group after hematoxylin-eosin staining was regular, and the groups A and B appeared with vacuolate cells, part of the structure disappeared, and the structures of acetabular and femoral head articular cartilage were complete,with less vacuole in the group C.(3) TUNEL method found that the apoptosis rate in the group C was lower than that in the group B, but higher than that in the control group(P < 0.05), indicating the decreased apoptotic rate after reduction.(4) Correlation analysis showed that the apoptosis rate was positively correlated with the expression level of Caspase-3 and negatively correlated with Bcl-2 expression.(5) To conclude, the reduction group shows a highly expressed Bcl-2, decreased expression of Caspase-3 and reduced apoptosis rate. Therefore,the reduction method can reduce the apoptosis rate of acetabular chondrocytes in developmental dysplasia of the hip, delay the degeneration of cartilage, and slow the course of disease progression.
引文
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