兔膝关节损伤后软骨细胞中OPG与RANKL表达

目的:随着中国经济的高速发展,人民生活水平日益增高。体育运动,高能量机动车造成的膝关节损伤数量不断增加,关节软骨的损伤恢复一直不是很理想。针对其后期的恢复研究做了许多相关的实验。本实验通过对兔膝关节软骨细胞损伤后软骨细胞中OPG(骨保护素)与RANKL(核激活因子受体配体)两种因子在损伤后与正常软骨细胞中的表达的比较,探讨损伤后软骨细胞中OPG和RANKL的表达。方法:25只大耳白兔随机分成5组,每组5只。前4组在10天完成20只兔左后腿膝关节软骨损伤动物模型,最后1组5只做手术对照组,术后处置相同。在术后1周,2周,4周,8周时取膝关节软骨损伤处关节软骨制成标本对照组取正常膝关节软骨制成标本。应用HE染色观察软骨细胞恢复情况及SP免疫组化法检测软骨细胞中OPG与RANKL的表达。在光镜下观察,通过医学图像分析软件(media cybernetics image-proplus6.0)图片分析,测定镜下相对灰度值并计算每个标本的平均灰度值,单因素方差分析法进行数据分析。结果:OPG在膝关节损伤后2-4周表达最高,4-8周有下降趋势。RANKL在膝关节软骨损伤后明显表达并逐步的呈升高趋势。实验组镜下观察表达较强烈,测定镜下平均灰度值与对照组有显著性差异(P0.05)。结论:1.OPG在膝关节软骨损伤后显著性表达。2-4周表达较高,4-8周趋于降低。2.RANKL在膝关节软骨损伤后显著表达,1-8周呈平稳上升趋势。

The Expressions of OPG and RANKL in the Rabbit Injured Knee JointCartilage Cells

Objective:With the rapid development of Chinese economy, the people''s living standards are increasing. The num- berof knee injuries caused by doing sports and high-energy motor vehicle is increasing. But the restoration of the articular cartilage has notbeen very satisfactory. So a lot of research-related experiments were used to analyze their recovery in the current study. By comparing theexpression of the two factors of OPG (osteoprotegerin) and RANKL (Ligand of receptor activator of NF-kbeta, RANKL) between the injuryand normal cartilage cells, we investigate the distribution of OPG and RANKL content of rabbit knee joint cartilage injury cells in differentperiods.Methods:25 rabbits were randomly divided into five groups. There were five rabbits in each group. Among the five groups,the four groups were used to complete the animal models of 20 rabbits left hind knee cartilage injury during 10 days, and another onegroup were used to control group under the same disposal surgery. The rabbits in the first four groups were killed after 1 week, 2 weeks, 4weeks, and 8 weeks, respectively. At the same time, the knee injury cartilages were made as the specimen to observe the structure. Therabbits of control group were killed in appropriate time when the rabbits were healed after surgery and stuffed knee cartilage. The recoveryof Chondrocytes was observed by HE staining. The expression of the OPG and RANKL in chondrocytes were detected with SP immunohistochemicalmethod. The medical image analysis software (media cybernetics image-proplus6.0) was used to analyze Image, calculatethe relative gray value and the average gray value of each specimen under the light microscope; ANOVA was used to data analysis.Results:OPG expression was highest in 2-4 weeks after injury, while it showed a decrease trend in 4-8 weeks. RANKL expression afterknee cartilage injury was clear and gradually increasing, which may have some correlation with the heal shaping process of cartilageinjury. At the same time, more intense expression were observed in the experimental group. There were significant difference of the averagegray value between the experimental group and the control group (P<0.05).Conclusion:1.OPG expression was significant after kneecartilage injury. Higher expression appeared in 2-4 weeks. It shows decreasing trend in the 4-8 weeks. 2. RANKL expression was alsosignificant, which showed steady upward trend in the 1-8 weeks.