应力刺激下TNF-α在大鼠跟腱末端区病变过程中的表达

发布时间：2018-02-14 05:55

本文关键词： 末端病 TNF-α NF-κB　出处：《苏州大学》2014年硕士论文　论文类型：学位论文

【摘要】：研究目的：人们从分子水平进行了大量的有关TNF-α与NF-κB信号通路以及两者之间相互关系的研究，但对于在末端病发生过程中作用的研究还处于探索阶段，直接相关研究很少。TNF-α作为重要的炎症因子，可单独或通过与其他细胞因子的相互作用引起促进炎症反应。NF-κB信号通路从多方面参与末端病的形成，但这是一个复杂的细胞因子网络，其具体调控及反馈调节机制、细胞因子网络的交互作用、信号通路间的可能联系尚有待逐步揭示。因而进一步加强对TNF-α介导的NF-κB信号转导途径的研究，尤其是TNF-α和NF-κB抑制剂进行末端病治疗，可能为延缓或阻断末端病进程提供新的尝试，并为末端病的早期预防和临床治疗提供理论依据研究方法：本实验通过大负荷跳跃运动方式建立末端病损伤模型，从炎症因子TNF-α与NF-κB入手，探讨末端病发生的分子机制。研究方法：48只8周龄雄性SD大鼠，购买于苏州大学动物实验中心。大鼠随机分为对照组和跳跃组，，对照组24只，跳跃组24只。对照组正常环境下饲养，跳跃组放于半自动跳跃电刺激笼内进行为期6周的跳跃运动。每两周取材一次，大鼠麻醉处死，取大鼠双后足跟腱及近端跟骨区域，一组材料利用免疫组化法检测末端组织内TNF-α和NF-κB的蛋白含量，另外一组制作HE石蜡切片并染色观察。观察比较三组实验组和对照组大鼠的跟腱末端区的病理组织变化，分析不同组大鼠的TNF-α、NF-κB的表达情况。研究结果： (1)HE染色结果表明，跳跃组与对照组病理组织形态出现较大差异，且随着造模时间延长，差异明显增大。 (2)对照组2、4、6周大鼠肌腱、纤维及钙化软骨和跟骨TNF-α阳性细胞数无显著性差异（p＞0.05），与对照组相比，跳跃2、4、6周组大鼠肌腱、纤维及钙化软骨和跟骨TNF-α阳性细胞数显著增加，具有非常显著性差异（P＜0.01）。 (3)对照组2、4、6周大鼠肌腱、纤维及钙化软骨和跟骨NF-κ B阳性细胞数无显著性差异（p＞0.05），与对照组相比，跳跃2、4、6周组大鼠在肌腱、纤维及钙化软骨和跟骨NF-κB阳性细胞数增加显著，，具有非常显著性差异（P＜0.01）。研究结论： (1)TNF-α和NF-κB对应力刺激敏感，TNF-α和NF-κB表达升高，是末端病产生的机制之一。 (2)在末端区各区TNF-α和NF-κB的含量不同，其中肌腱和纤维及钙化软骨区偏高，骨区则偏低。
[Abstract]:Objectives of the study:. A lot of studies have been carried out on TNF- 伪 and NF- 魏 B signaling pathway and their relationship at molecular level, but the role of TNF- 伪 and NF- 魏 B in the process of terminal disease is still in the exploratory stage. As an important inflammatory factor, there are few direct correlation studies. TNF- 伪 can promote inflammatory response. NF- 魏 B signaling pathway is involved in the formation of terminal disease in many ways, either alone or through interaction with other cytokines. But this is a complex cytokine network, and its specific regulation and feedback regulation mechanisms, the interaction of cytokine networks, The possible relationship between signal pathways has yet to be gradually revealed. Therefore, the study of TNF- 伪 -mediated NF- 魏 B signal transduction pathway, especially the treatment of terminal diseases by TNF- 伪 and NF- 魏 B inhibitors, has been further strengthened. It may provide a new attempt to delay or block the progression of terminal diseases, and provide theoretical basis for early prevention and clinical treatment of terminal diseases. Research methods:. In this experiment, the model of terminal disease injury was established by means of high-load jumping exercise, and the molecular mechanism of terminal disease was discussed from the inflammatory factors TNF- 伪 and NF- 魏 B. Methods Forty eight 8-week-old male Sprague-Dawley rats were purchased from Suzhou University Animal experiment Center. The rats were randomly divided into two groups: control group (n = 24) and jumping group (n = 24). The jumping group was placed in a semi-automatic jumping electric stimulation cage for 6 weeks. The rats were anesthetized and killed every two weeks. The Achilles tendon and the proximal calcaneal area of the rats were harvested. One group of materials was used to detect the protein content of TNF- 伪 and NF- 魏 B in the terminal tissues by immunohistochemical method, the other group was made of HE paraffin sections and stained. The pathological changes of the ends of the Achilles tendon in the experimental group and the control group were observed and compared. To analyze the expression of TNF- 伪 and NF- 魏 B in different groups of rats. Results of the study:. The results of HE staining showed that the histopathology of the jumping group and the control group were significantly different, and the difference increased with the prolongation of the model making time. (2) there was no significant difference in the number of TNF- 伪 positive cells in tendon, fiber, calcified cartilage and calcaneal bone of rats in the control group at 6 weeks (P > 0.05). Compared with the control group, the number of TNF- 伪 positive cells in tendon, fiber and calcified cartilage and calcaneal bone were significantly increased in the control group. There was significant difference (P < 0.01). (3) there was no significant difference in the number of NF- 魏 B positive cells in tendon, fiber and calcified cartilage and calcaneal bone of rats in the control group at 6 weeks, and the number of NF- 魏 B positive cells in the control group was not significantly different from that in the control group (P > 0.05). The number of NF- 魏 B positive cells in fibrous and calcified cartilage and calcaneus increased significantly (P < 0.01). The study concluded that:. The increased expression of TNF- 伪 and NF- 魏 B is one of the mechanisms of terminal diseases. (2) the contents of TNF- 伪 and NF- 魏 B in the terminal region were different, in which the tendons and fibers and calcified cartilage were higher and the bone areas were lower.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：G804.2

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