跳跃负荷致髌骨髌腱结合部和髌腱纤维化机制及冷水浴干预影响

发布时间：2018-08-30 18:18

【摘要】：目的:通过电刺激定量跳跃装置使兔完成相应的跳跃训练时长,探索髌骨髌腱结合部和髌腱纤维化进程机制,并探究冷水浴干预对髌骨髌腱结合部纤维化的影响。方法:将18周龄雌性新西兰兔随机分为对照组、跳跃组、运动后冷水浴组和单纯冷水浴组,各组又分为2、4、6、8周4个时长。跳跃组完成每天150次,每周5天的跳跃训练;运动后冷水浴组每天跳跃训练完成后进行3次,每次5min的间歇冷水浴;单纯冷水浴组完成相同的冷水浴任务。在对应时长取下髌骨髌腱结合部(PPTJ)和髌腱(PT)进行检测。采用HE、番红、马松、免疫组织化学和免疫组织荧光染色及Real Time PCR技术,描述评价跳跃和跳跃后冷水浴干预对PPTJ和PT组织学变化及纤维化和抗纤维化相关因子表达的影响。结果:(1)与对照组同时间点相比,跳跃组PPTJ所有时间点的FZT、GA、CA、TGF-β1、Smad3、CTGF、COLI、COLIII显著升高(p0.01);跳跃4周组PT的TGF-β1、CTGF、COLI、COLIII mRNA及蛋白显著增高(p0.05)。跳跃组内各时间点相比,PPTJ的6周组FZT、GA、CA、TGF-β1、Smad3、CTGF、COLI最高(P0.05);PT的4周组TGF-β1、COLI、COLIII mRNA及蛋白最高(p0.01)。(2)与跳跃组同时间点相比,运动后冷水浴组PPTJ的FZT、GA、CA、TGF-β1、Smad3、CTGF、TIMP-1、COLI、COLIII 4周、6周组显著下降(P0.05);HGF、MMP-1、MMP-9 4周组显著增加(P0.01)。(3)与对照组和单纯冷水浴组相比,运动后冷水浴组PPTJ纤维化因子表达显著升高(P0.01),抗纤维化因子表达显著降低(P0.01),单纯冷水浴组与对照组无差异(P0.05)。结论:(1)跳跃负荷导致PT纤维化峰值期早于PPTJ出现,PT出现于4周,PPTJ在6周。(2)跳跃负荷致PPTJ和PT纤维化机制为增加TGF-β1和Smad3分泌,上调CTGF表达,促使COLI、COLIII合成增加,胶原纤维面积增大,致使发生纤维化改变。(3)单纯冷水浴对PPTJ组织无影响,运动后冷水浴可以改善PPTJ纤维化,且在4周效果显著。(4)运动后冷水浴可能通过两条途径抑制PPTJ纤维化进程,一是下调TGF-β1和Smad3表达,抑制后续信号通路;二是促进HGF分泌,促使TIMP-1与MMP-1、MMP-9恢复平衡,降低COLI、COLIII合成,减少胶原纤维面积,抑制纤维化进程。
[Abstract]:AIM: To explore the mechanism of fibrosis of patellar tendon junction and patellar tendon, and to explore the effect of cold water bath on fibrosis of patellar tendon junction. Methods: Female New Zealand rabbits aged 18 weeks were randomly divided into control group, jumping group, cold water bath group and post-exercise cold water bath group. Each group was divided into 2,4,6,and 8 weeks with 4 time lengths.The jumping group completed 150 times a day and 5 days a week jumping training.The cold water bath group was given intermittent cold water bath 3 times a day and 5 minutes each time after exercise.The cold water bath group completed the same cold water bath task.The patellar patellar tendon junction (PPTJ) and patellar tendon junction (PPTJ) were removed in corresponding time. Patellar tendon (PT) was examined. HE, saffron, masson, immunohistochemistry, immunohistochemistry, immunohistofluorescence staining and Real Time PCR were used to describe and evaluate the effect of cold water bath intervention after jumping and jumping on the histological changes of PPTJ and PT, and the expression of fibrosis and anti-fibrosis related factors. FZT, GA, CA, TGF-beta 1, Smad3, CTGF, COLI, COLIII were significantly increased at time points (p0.01); TGF-beta 1, CTGF, COLI II mRNA and protein were significantly increased in the four-week skipping group (p0.05). Compared with each time point in the skipping group, FZT, GA, CA, TGF-beta 1, Smad3, CTGF and COLI were the highest in the six-week group of PPTJ (p0.05); TGF-beta 1, COLI 2, COLI, COLI II and protein were the highest in the four-week skipping group of PT (p0.05). (2) Compared with the jumping group, FZT, GA, CA, TGF-beta 1, Smad3, CTGF, TIMP-1, COLI, COLIII 4 weeks, 6 weeks group decreased significantly (P 0.05); HGF, MMP-1, MMP-9 4 weeks group increased significantly (P 0.01). (3) Compared with the control group and the simple cold water bath group, the expression of PPTJ fibrosis factor in the cold water bath group increased significantly (P 0.01). Conclusion: (1) The peak of PT fibrosis appeared earlier than PPTJ, PT appeared in 4 weeks and PPTJ appeared in 6 weeks. (2) The mechanism of PPTJ and PT fibrosis was to increase the secretion of TGF-beta 1 and Smad3, up-regulate the expression of CTGF and promote the synthesis of COLI and COLIII. (3) Cold water bath alone has no effect on PPTJ tissue, and cold water bath after exercise can improve PPTJ fibrosis, and the effect is significant in 4 weeks. (4) Cold water bath after exercise may inhibit PPTJ fibrosis process through two ways, one is to down-regulate the expression of TGF-beta 1 and Smad3, inhibit the subsequent signaling pathway; Secondly, it can promote the secretion of HGF, restore the balance between TIMP-1 and MMP-1, MMP-9, reduce the synthesis of COLI and COLIII, reduce the area of collagen fibers and inhibit the fibrosis process.
【学位授予单位】：北京体育大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：G804.2

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