低强度脉冲超声干预膝关节炎模型兔软骨细胞Ⅱ型胶原和基质金属蛋白酶13的表达

Effects of low intensity pulsed ultrasound on the expression of type II collagen and matrix metalloproteinase-13 in chondrocytes of a rabbit model of knee arthritis

背景:低强度脉冲超声安全无创、穿透性强,近年来,越来越多的研究表明,低强度脉冲超声在促进损伤关节软骨修复方面具有重要的作用。目的:观察低强度脉冲超声对膝关节炎模型兔软骨细胞Ⅱ型胶原和基质金属蛋白酶13表达的影响。方法:纳入20只新西兰白兔,均建立膝关节炎模型,建模后随机分为对照组和实验组,每组10只。实验组动物右侧膝关节利用超声波骨折愈合仪进行低强度脉冲超声治疗,对照组动物予以右侧膝关节假低强度脉冲超声治疗。干预后8周,取胫骨内侧端以及膝关节股骨内侧端软骨组织,经常规处理后,进行甲苯胺蓝染色,并置于显微镜下进行病理学观察,利用q RT-PCR法软骨细胞基质金属蛋白酶13和Ⅱ型胶原表达量检测。观察实验动物一般情况和关节软骨病理形态,以及基质金属蛋白酶13、Ⅱ型胶原的q RT-PCR检测结果。结果与结论:1甲苯胺蓝染色和病理学观察:治疗8周后,对照组关节软骨出现明显的裂隙,并从表面延伸至深层,且存在严重的染色缺失现象;实验组可观察到关节软骨表面形成裂隙,存在中度染色缺失现象;2q RT-PCR检测:软骨细胞培养5,10 d后,实验组和对照组相同时间点基质金属蛋白酶13和Ⅱ型胶原基因表达差异均有显著性意义(P<0.05);3结果证实:低强度脉冲超声可调节膝关节炎模型兔软骨细胞Ⅱ型胶原和基质金属蛋白酶13基因的表达。

BACKGROUND： Low intensity pulsed ultrasound is safe, non-invasive, and has strong penetration. In recent years, more and more studies have indicated that low intensity pulsed ultrasound has important role in promoting the repair of damaged articular cartilage. OBJECTIVE： To investigate the effect of low intensity pulsed ultrasound on the expression of type II col agen and matrix metal oproteinase 13 in articular cartilage of rabbits of knee arthritis. METHODS： A total of 20 New Zealand white rabbits were included in this study to establish knee arthritis model. Al models were randomly divided into control group and experimental group, 10 rats in each group. In the experimental group, the right knee joint received low intensity pulsed ultrasound treatment with an ultrasonic fracture healing instrument. In the control group, right knee joint received false low intensity pulsed ultrasound therapy. At 8 weeks after intervention, cartilage tissue was col ected from medial end of tibia and medial end of femur. After conventional treatment, toluidine blue staining was performed. Pathological observation was conducted under a microscope. Quantitative real-time polymerase chain reaction was used to detect matrix metal oproteinase 13 and type II col agen expression in chondrocytes. The general situation of the experimental animals was observed. The results of articular cartilage and the quantitative real-time polymerase chain reaction results of matrix metal oproteinase-13 and type II col agen were observed under a microscope. RESULTS AND CONCLUSION： （1） Toluidine blue staining and pathological observation： 8 weeks after treatment, obvious fissure appeared in articular cartilages, and extending from the surface to the deep layer in the control group, and there was a lack of staining. In the experimental group, cracks formed on articular cartilage surface, and there was a moderate staining missing. （2） Quantitative real-time polymerase chain reaction： at 5 and 10 days after culture, significant differences in gene expression of matrix metal oproteinase-13 and type II col agen were detected at various time points in the experimental and control groups （P 〈 0.05）. （3） The results showed that the low intensity pulsed ultrasound could affect the gene expression of matrix metal oproteinase-13 and type II col agen in the rabbit model of knee arthritis.