文献类型：期刊文献

中文题名：关节腔注射联合跑台运动建立大鼠慢性骨关节炎模型

英文题名：Establishment of a rat model of chronic osteoarthritis by intra-articular injection combined with treadmill exercise

作者：陈世杰[1,2];朱杨雄[1,2];劳文艳[1,2,3];周艳丽[1,2];赵晓红[1,2,3]

第一作者：陈世杰

机构：[1]北京联合大学生物化学工程学院;[2]北京联合大学功能食品科学技术研究院;[3]北京联合大学生物活性物质与功能食品北京市重点实验室

第一机构：北京联合大学生物化学工程学院

年份：2018

卷号：22

期号：24

起止页码：3849-3856

中文期刊名：中国组织工程研究

外文期刊名：Chinese Journal of Tissue Engineering Research

收录：CSTPCD;;北大核心:【北大核心2017】；

基金：北京联合大学生物活性物质与功能食品北京市重点实验室开放课题(Zk70201501)~~

语种：中文

中文关键词：骨关节炎;模型;关节软骨;滑膜;跑台运动;强度;组织病理学;细胞因子;蛋白聚糖;Ⅱ型胶原;组织构建

外文关键词：Osteoarthritis;Cytokines;Proteoglycans;Collagen Type Ⅱ;Tissue Engineering

摘要：背景:建立骨关节炎动物模型是研究骨关节炎病理机制、预防及治疗的重要手段。目的:进一步验证关节腔注射联合跑台运动建立大鼠慢性骨关节炎模型的可行性。方法:大鼠随机分为安静组、低强度组(跑台坡度0°,速度15 m/min)、中强度组(坡度5°,速度20 m/min)和高强度组(坡度10°,速度27 m/min),每组6只。实验开始后,采用关节腔注射白陶土-鹿角菜胶诱发大鼠膝关节急性损伤,术后第3天各运动组大鼠在跑台上以慢速跑步适应跑道,5 d后开始正式实验,每天运动1 h,每周休息1 d,持续6周。每周定期测量大鼠关节肿胀度,动物处死后,取膝关节制作石蜡组织切片,进行苏木精-伊红和番红-O-固绿染色,观察关节软骨组织病理学变化,取血清和滑膜,ELISA法测定滑膜基质金属蛋白酶3、基质金属蛋白酶组织抑制剂1、白细胞介素1β水平,黄嘌呤氧化酶法测定血清超氧化物歧化酶含量,以及免疫组化检测关节软骨Ⅱ型胶原的表达情况。结果与结论:(1)与安静组比较,中、高强度组大鼠膝关节组织病理学评分均明显增高(P<0.05或P<0.01);关节肿胀度明显增大(P<0.01或P<0.05);滑膜基质金属蛋白酶3、基质金属蛋白酶组织抑制剂1和白细胞介素1β含量明显增高(P<0.01),基质金属蛋白酶组织抑制剂1/基质金属蛋白酶3浓度比值显著降低(P<0.01);血清超氧化物歧化酶活性和关节软骨Ⅱ型胶原平均光密度值显著降低(P<0.01);而低强度组除大鼠关节软骨Ⅱ型胶原平均光密度值明显降低(P<0.05)外,其余指标均无明显变化;(2)中、高强度组各项指标变化与低强度组差别显著(P<0.01或P<0.05);(3)高强度组除大鼠滑膜基质金属蛋白酶3含量明显高于中强度组(P<0.01)及基质金属蛋白酶组织抑制剂1/基质金属蛋白酶3浓度比值明显低于中强度组(P<0.05)外,其余指标与中强度组差别不显著;(4)结果表明,中、高强度运动可加重大鼠关节肿胀损伤、组织病理学异常征象,加重大鼠氧化损伤及炎性损伤,同时加速关节软骨基质降解,从而造成大鼠慢性骨关节炎产生,结合实际操作情况,中等强度运动为建立大鼠慢性骨关节炎模型的最佳运动条件。
BACKGROUND： Establishment of an animal model of osteoarthritis is an important means to study the pathogenesis, prevention and treatment of osteoarthritis. OBJECTIVE： To further verify the feasibility of establishing a rat model of osteoarthritis by intra-articular injection combined with treadmill exercise. METHODS： The rats were randomly divided into sedentary, low-intensity（incline 0° and speed 15 m/min）, moderate-intensity（incline 5° and speed 20 m/min） and high-intensity（incline 10° and speed 27 m/min） groups（n=6 per group）. The acute injury of the knee joint was induced by injecting kaolin and carrageen-λ into the articular cavity. At 3 days after modeling, the adaptive exercise was conducted at a low speed, and 5 days later the formal training was conducted, 1 hour per day and 1 day off every week for continuous 6 weeks. The degree of joint swelling was measured once a week. The rats were sacrificed after 6 weeks of exercise and the paraffin continuous sections of knee joints were prepared and stained with hematoxylin and eosin, and safranin 0 and fast green to observe the pathological changes of articular cartilage. The contents of matrix metalloproteinase-3, tissue inhibitor of metalloproteinases 1, and interleukin-1β in synovial tissues were detected with ELISA kits. The content of superoxide dismutase in serum was measured by xanthine oxidase method, and the expression of collagen type II was detected by immunohistochemistry. RESULTS AND CONCLUSION： Compared with the sedentary group, the histopathological scores were significantly increased in the moderate-, and high-intensity groups（P〈0.05 or P〈0.01）; the degree of joint swelling was significantly increased（P〈0.05 or P〈0.01）; the contents of matrix metalloproteinase-3, tissue inhibitor of metalloproteinases 1, and interleukin-1β in synovial tissues were significantly up-regulated（P〈0.01）, and the tissue inhibitor of metalloproteinases 1/matrix metalloproteinase-3 ratio was significantly reduced（P〈0.01）; the level of superoxide dismutase in serum and the average optical density value of collagen type II in the articular cartilage were significantly decreased（P〈0.01）. The average optical density value of collagen type II in the low-intensity group was significantly reduced（P〈0.05）, but there was no significant change in the other indexes. The changes in the indexes in the moderate-and high-intensity groups were significantly different from those in the low-intensity group（P〈0.01 or P〈0.05）. In the high-intensity group, the content of matrix metalloproteinase-3 in serum was significantly higher（P〈0.01） and the tissue inhibitor of metalloproteinases 1/matrix metalloproteinase-3 ratio in the synovium was significantly lower（P〈0.05） compared with the moderate-intensity group, but there was no significant difference in the other indexes between the two groups. These results indicate that the moderate-and high-intensity exercise can aggravate joint swelling and histopathological abnormalities, aggravate oxidative and inflammatory damage and accelerate cartilage degradation in rats, eventually resulting in chronic osteoarthritis. According to the actual operation, the moderate-intensity exercise is the best choice for establishing the chronic osteoarthritis model in rats.