中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (24): 3849-3856.doi: 10.3969/j.issn.2095-4344.0821

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

关节腔注射联合跑台运动建立大鼠慢性骨关节炎模型

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   

  1. 北京联合大学，1生物化学工程学院，2功能食品科学技术研究院，3生物活性物质与功能食品北京市重点实验室，北京市  100191
  • 收稿日期:2017-11-01
  • 通讯作者: 赵晓红，博士，研究员，北京联合大学生物化学工程学院, 功能食品科学技术研究院，生物活性物质与功能食品北京市重点实验室，北京市 100191
  • 作者简介:陈世杰，女，1991年生，河南省宝丰县人，汉族，2017年北京联合大学大学毕业，硕士，主要从事生物活性物质的生理功能研究。
  • 基金资助:

    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)

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

Chen Shi-jie1, 2, Zhu Yang-xiong1, 2, Lao Wen-yan1, 2, 3, Zhou Yan-li1, 2, Zhao Xiao-hong1, 2, 3   

  1. 1College of Biochemical Engineering, 2Institute for Science and Technology of Functional Food, 3Beijing Key Laboratory of Bioactive Substances and Functional Foods, Beijing Union University, Beijing 100191, China
  • Received:2017-11-01
  • Contact: Zhao Xiao-hong, Ph.D., Researcher, College of Biochemical Engineering, Beijing Union University, Beijing 100191, China
  • About author:Chen Shi-jie, Master, College of Biochemical Engineering, Beijing Union University, Beijing 100191, China; Institute for Science and Technology of Functional Food, Beijing Union University, Beijing 100191, China
  • Supported by:

    the Open Project of Beijing Key Laboratory of Bioactive Substances and Functional Foods of Beijing Union University, No. Zk70201501

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摘要：

文章快速阅读：
文题释义：
蛋白多糖：关节软骨是一种特殊类型的结缔组织，由细胞外基质(extracellularmatrix，EMC)和软骨细胞组成，软骨基质中蛋白质与多糖以共价键和非共价键相连构成的多种巨大分子称为蛋白多糖(proteoglycans，PG)。蛋白多糖分子能吸附大量水分，使软骨具有承重作用。当关节软骨受到压力时，软骨中的水分受压流失，而当压力消失时，水分又被重新吸收。因此，蛋白多糖决定了软骨的压力特性，使软骨组织具有延展性和抗压缩能力。
基质金属蛋白酶：是一类广泛存在于机体各种结缔组织中的蛋白酶家族，在细胞外基质代谢的调节中有着非常重要的作用。基质金属蛋白酶是一种水解所有细胞外基质的蛋白水解酶，被认为是机体病理破坏和生理重建的主要基础因素之一，在关节软骨损伤和退变中也起着很重要的作用。
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。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程
ORCID: 0000-0001-6347-7656(陈世杰)

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

Abstract:

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.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoarthritis, Cytokines, Proteoglycans, Collagen Type II, Tissue Engineering

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