抗炎干预对兔髌腱及止点结构及细胞因子变化的影响

发布时间：2018-06-24 20:24

  本文选题：髌腱 + 腱止点　； 参考：《北京体育大学》2017年博士论文

【摘要】：目的:观察不同周数跳跃训练及抗炎干预对兔髌腱及腱止点组织学,免疫组化,实时荧光定量PCR等指标的变化,探讨负荷积累量与抗炎干预对髌腱及腱止点结构变化、炎症反应相关蛋白、细胞外基质合成与降解相关蛋白及自噬相关蛋白的影响,观察布洛芬抗炎药预防腱病的效果,并揭示其作用机理。方法:68只雌性新西兰大白兔(体重2.46±0.11kg)随机分为4组,即空白对照组CON,跳跃训练组OJ,跳跃加抗炎药组JAD以及抗炎药组OAD,各组又按时间不同分为2周,4周,6周及8周组。其中研究一仅涉及腱止点,研究二仅涉及髌腱,研究三涉及二者之间的关系。跳跃训练采用三维测力台记录起跳、落地峰值力及冲量,分别于对应的时间点取材,腱止点标本行HE、番红及免疫组化染色,髌腱行免疫组化及RT-PCR。观察指标包括纤维软骨带厚度、细胞密度,细胞因子TNFα、HMGB1、MMP-1、MMP-2、Beclin-1、LC3B、IL-1β、COX-2、PTGES2、COL-I、COL-III;其中腱止点检测所有指标的蛋白表达,髌腱样本仅检测后5个指标的蛋白及mRNA表达量。结果:(1)不同周数跳跃训练后经体重标化的生物力学结果显示,OJ4W组的起跳冲量显著低于OJ2W组(P=0.005);(2)组织学结果显示,与相同时间点的CON组相比,OJ组随跳跃训练周数的变化依次呈现出以下变化规律:腱止点处细胞密度增大,纤维软骨带增厚,潮线模糊或消失,细胞形态改变,分布不均,骨腱结合部固有的分层结构消失,偏振光下胶原波浪形结构消失、排列不连续;番红染色下粘多糖蛋白异常增多维持在一定的水平上;(3)免疫组化结果显示,与相同时间点的CON组相比,OJ2W、OJ4W组TNFα、IL-1β、COX-2、PTGES2、MMP2显著升高(P0.05),OJ6W组IL-1β、COX-2、MMP2、COL-III显著升高(P0.05),OJ8W组IL-1β、COX-2、COL-I显著升高但MMP1、Beclin显著降低(P0.05);与同时间点的OJ组相比,JAD2W组COX-2、PTGES2、MMP2显著降低(P0.05),JAD6W组MMP1、COL-I、COL-I/COL-III、LC3B抗炎干预对兔髌腱及止点损伤形成的影响显著降低(P0.05),JAD8W组HMGB1、MMP1、COL-I、LC3B显著降低(P0.05)。结论:(1)髌腱及止点结构、炎症因子变化关系密切。TNFα、IL-1β、MMPs、COL-I、COL-III等蛋白表达具有一定的时序性规律。(2)早期存在一定程度的炎症反应是有益的,而持续存在的炎症反应是对机体不利的。(3)抗炎干预具有较好的短期效应,集中体现在早期有助于缓解炎细胞浸润及保护胶原蛋白,但长期效应来看,仅能提前机体修复反应的时间点,对最终的愈合质量帮助不大。(4)髌腱对载荷的承载能力明显大于腱止点,可以解释腱止点损伤多于肌腱的原因。
[Abstract]:Objective: to observe the changes of histological, immunohistochemical and real-time fluorescent quantitative PCR in rabbit patellar tendon and tendon after different weeks of jumping training and anti-inflammatory intervention, and to investigate the changes of load accumulation and anti-inflammatory intervention on patellar tendon and tendon insertion. The effects of ibuprofen anti-inflammatory drugs on the prevention of tendon disease were observed and the mechanism of the effects of ibuprofen anti-inflammatory drugs on the prevention of tendonitis was observed in order to investigate the effects of inflammatory response related proteins, extracellular matrix synthesis and degradation related proteins and autophagy related proteins. Methods Sixty-eight female New Zealand white rabbits (weight 2.46 卤0.11kg) were randomly divided into four groups: control group (control group), jumping training group (OJ), jumping plus anti-inflammatory drug group (JAD) and anti-inflammatory drug group (OAD). Each group was divided into 2 weeks, 4 weeks, 6 weeks and 8 weeks groups. Among them, one involves tendon insertion, the other involves patellar tendon, and the third concerns the relationship between the two. The jump was recorded by a three dimensional force measuring table. The peak force and impulse were recorded at the corresponding time point. The specimens were collected at the corresponding time point. The specimens were stained with HEH, Phanerin and immunohistochemistry, and the patellar tendon was stained with immunohistochemistry and RT-PCR. The observed parameters included the thickness of fibrochondrocytes, cell density, cytokine TNF 伪, HMGB1, MMP-1, MMP-2, Beclin-l, LC3BU, IL-1 尾, COX-2PTGES2PTGES2, COL-ICOL-III, in which the protein expression of all the indexes was detected at the tendon insertion point, and the protein and mRNA expression in the patellar tendon samples was detected only for the last five indexes. Results: (1) after different weeks of jumping training, the biomechanical results showed that the take-off impulse of OJ4W group was significantly lower than that of OJ2W group (P0. 005); (2). Compared with the Con group at the same time point, the changes of the number of jump training weeks in the OJ group showed the following changes in turn: the cell density at the tendon stop point increased, the fibrous cartilage band thickened, the tidal line blurred or disappeared, and the cell morphology changed unevenly. The lamellar structure of the bone tendon disappeared, the collagen wavy structure disappeared under polarized light, and the abnormal increase of mucopolysaccharide protein was maintained at a certain level. (3) Immunohistochemical results showed that the abnormal increase of mucopolysaccharide protein was maintained at a certain level. 涓庣浉鍚屾椂闂寸偣鐨凜ON缁勭浉姣，

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