【作者】 郭佳佳；

【导师】 张玉芹；

【作者基本信息】 首都体育学院 ， 运动人体科学， 2010， 硕士

【摘要】 目的采用“电击跳跃法”建立大鼠跟腱末端病模型,并对模型进行火针治疗。观察末端区组织和细胞形态变化、HSP70表达和细胞凋亡指数变化,依据末端结构特点从病因、病理角度探讨火针治疗末端病的作用机理,为末端病的火针疗法提供理论依据。方法以1月龄雄性SD大鼠为研究对象,通过“电击跳跃法”建立跟腱末端病模型。将88只大鼠随机分为3大组,分别为：正常组,治疗组(治疗1、2、3、4、5组)和对照组(对照1、2、3、4、5组)。造模后进行火针治疗,于1次治疗后第1、2、3、4天分别处死治疗1、2、3、4组及其相应对照组,再于4次治疗后第4天处死治疗5组和对照5组,取每只大鼠单侧后足跟腱和完整的跟腱末端区组织,经固定、脱钙、包埋、切片、贴片、烤片后,常规HE染色,观察末端区组织和细胞形态变化；采用免疫组化ABC法检测末端区跟腱、纤维软骨区、钙化软骨区、跟骨、腱骨关节面软骨的HSP70阳性率；应用TUNEL法检测跟腱、纤维软骨区、钙化软骨区、跟骨、腱骨关节面软骨、腱围的细胞凋亡指数(AI)。所有数据用SPSS13.0进行统计分析。结果(1)HE染色镜下观察：经1次火针治疗后,治疗1、2、3组大鼠跟腱末端区与对照组相比,在病理上无明显变化。治疗4组在腱围、跟腱等部位的病理有所改善,但差异并不显著。而经4次火针治疗后,治疗5组从病理上看明显减轻,更接近于正常组。(2)HSP70表达：1次火针治疗后,治疗组大鼠末端区主要在纤维软骨区、腱骨关节面软骨两个部位的HSP70阳性率比对照组有不同程度升高；经4次火针治疗后,纤维软骨区、腱骨关节面软骨两个部位的HSP70阳性率明显高于对照组；4次火针治疗后在纤维软骨区、腱骨关节面软骨两个部位的HSP70阳性率明显高于1次火针治疗。(3)细胞凋亡检测：1次火针治疗后,治疗组大鼠末端区主要在跟腱、纤维软骨区、腱围这三个部位的AI比对照组有不同程度降低；经4次火针治疗后,跟腱、纤维软骨区、腱骨关节面软骨、腱围的AI明显低于对照组；4次火针治疗后在跟腱、纤维软骨区、腱骨关节面软骨、腱围的AI明显低于1次火针治疗。结论(1)8周“电击跳跃法”造模训练成功地建立了大鼠跟腱末端病模型。(2)1次火针治疗诱导末端区合成大量HSP70,主要表现在纤维软骨区、腱骨关节面软骨两个部位,且在针刺后当天即达到一个高峰,之后3天呈连续下降趋势。4次火针治疗后这两个部位的HSP70合成量大大增加,HSP70阳性率明显升高,4次火针治疗能更进一步促进末端病的恢复。(3)1次火针治疗加速了跟腱、纤维软骨区、腱围三个部位凋亡细胞的吸收和清除,促进了末端区组织和细胞的修复。而4次火针治疗后,在跟腱、腱骨关节面软骨、纤维软骨区、腱围等多个部位的细胞凋亡指数都发生了改变,末端区的整体结构得到了恢复,4次火针治疗在促进末端病的整体恢复上效果更好。更多还原

【Abstract】 Purpose Establish the rat model of Achilles Tendon’s enthesiopathy by "electric shock jump".Then treat the model rats by using heated needling.By observing the terminal region of cell and tissue morphology, expression of HSP70 and change of cell apoptosis index, discuss the mechanisiam of heated needling therapy on enthesiopathy from a pathological point of view so as to provide a theoretical basis for heated needling therapy.Methods Take 1-month-old male SD rats as the object for study. By "electric shock jump" establish the rat model of Achilles Tendon’s enthesiopathy. The 88 rats were randomly divided into 3 groups, namely:the normal group, treatment group (treatment group 1,2,3,4,5) and control group (control group 1,2,3,4,5).After modeling, treat the rats by heated needling.Then kill each group of rats and the corresponding control group at the 1st,2nd,3rd,4th day after the first treatment and the last group and its control group at the fourth day after four times treatment. Cut unilateral hind tendon terminal region of the Achilles tendon and complete organization,then fixed,decalcified,embedded,cut,patch and baked it. After conventional HE staining,observe the terminal region of cell and tissue morphology. Using immunohistochemistry of ABC to detect positive rate of HSP70 at T,UF,MF,C and CA. Apply TUNEL to detect the apoptosis index(AI)of T,UF,MF, C,CA and PT. All datas are analyzed with SPSS13.0.Results (1) HE staining microscopic observation:After one time of heated needling treatment,compared with the corresponding control group,terminal region of Achilles tendon of treat group 1,2,3 rats has no significant change in the pathology. The treat group 4 has largely improved at T,CA and other parts of the Achilles tendon in the pathology, but the difference was not significant. After treated by heated needling therapy for 4 times, treat group 5 significantly reduced from the pathological point of view, closer to the normal group. (2) The expression of HSP70:After 1 time heated needling therapy, HSP70-positive rate of the treatment group in the terminal region mainly at the parts of UF and CA increased in varying degrees than the control group;(2) After 4 times heated needling therapy, HSP70-positive rate of the treatment group at the parts of UF and CA was significantly higher than the control group. HSP70-positive rate of the treatment group at the parts of UF and CA after 4 times heated needling therapy was significantly higher than the 1 time heated needling therapy groups. (3) The detection of apoptosis:After 1 time heated needling therapy, AI of the treatment group in the terminal region mainly at the parts of T,UFand PT decreased in varying degrees than the control group; After 4 times heated needling therapy, AI of the treatment group at the parts of T,UF, CA and PT was significantly lower than the control group; AI of the treatment group at the parts of T,UF, CA and PT after 4 times heated needling therapy was significantly lower than the 1 time heated needling therapy groups.Conclusions (1) Training by "electric shock jump" for 8 weeks successfully established the rat model of Achilles Tendon’s enthesiopathy. (2)1 time heated needling therapy can cause the tissues and cells of tenninal region to induce a large number of synthesis of HSP70 mainly at the two parts of UF and CA, and reached a peak on the therapy day, then showed a continuous downward trend during 3 days after therapy.After 4 times heated needling therapy, the synthesis of HSP70 on the two parts largely increased and positive rate of HSP70 also increased,further enhanced the recovery of enthesiopathy. (3) 1 time heated needling therapy accelerated the purge and absorption of apoptosis cells at T,UF and PT, promoted some tissues and cells repairing. After 4 times heated needling therapy,the main parts of T,CA,UF and PT had taken place great changes.So it is better for the overall restoration.更多还原