针刀松解法对肩周炎家兔模型抗氧化能力影响的研究
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摘要
肩周炎全称为肩关节周围炎,以肩部弥漫性疼痛与肩关节活动障碍为主要特征,是多发生于中老年人的慢性肩部疾患,其病变范围可涉及到肩关节周围肌肉、肌腱、筋膜、韧带、滑膜囊、关节囊等软组织。目前对其具体发病机制尚不明了
研究目的：
肩周炎有明显的病理分期,目前的临床研究多认为针刀松解法对粘连期肩周炎治疗效果较好。本研究在建立家兔肩周炎模型的基础上,以针刀松解为干预手段,采用病理学和生物化学方法,从调节抗氧化能力方面寻求肩周炎的发病机制与针刀干预的起效机制,寻找针刀松解法干预肩周炎的最佳时机,为肩周炎的针刀临床治疗提供实验依据。
研究方法：
选用兔龄6±0.5个月的健康雄性新西兰家兔48只,每只体重2.5±0.5kg。随机分为6组,分别为正常组8只、模型组8只、电针组8只,针刀组A(疼痛期干预)8只、针刀组B(僵硬期干预)8只、针刀组C(疼痛期及僵硬期共同干预)8只。应用持续劳损加冰敷方法制备肩周炎家兔模型,针刀各组行针刀治疗,电针组行电针治疗,正常组、模型组,不干预,与治疗各组,同等条件下饲养,共33天。取材分别取血浆、肱二头肌、冈上肌、冈下肌、大圆肌,测定丙二醛水平、超氧化物歧化酶活力、总抗氧化能力、转化生长因子-β1水平,取肱二头肌肌腱起始处至远侧3-4mm肌腱和前下方3-4mm滑膜,固定、包埋切片,予HE染色后光镜下观察其病理改变。
实验结果：
1血浆及局部各肌肉组织的丙二醛、超氧化物歧化酶活力、总抗氧化能力的测定结果显示,模型组的丙二醛水平普遍高于正常组、电针组、针刀组A、针刀组B、针刀组C,且多数具有显著性的差异,而模型组的超氧化物歧化酶活力、总抗氧化能力测定结果要普遍低于正常组、电针组、针刀组A、针刀组B、针刀组C。
针刀组A、针刀组B、针刀组C血浆及局部各肌肉组织的抗氧化能力与脂质过氧化物水平,对比有些具有显著性差异,有些无显著性差异,但相对于模型组,以上三组的丙二醛水平相对较低,而超氧化物歧化酶活力、总抗氧化能力相对较高。
2局部病理组织观察显示,正常组：滑膜：滑膜细胞多为1层,有些部位也可见2-3层,在皱襞较多处滑膜细胞可达5-6层。滑膜细胞深层结构疏松,可见少量毛细血管。肌腱：外膜(腱鞘脏层)由结缔组织包绕,最外层上皮细胞为立方细胞1-2层,排列整齐,腱纤维排列整齐。
模型组：滑膜：滑膜细胞多为1-2层,滑膜下毛细血管较多,管腔内可见较多红细胞。肌腱：外膜上皮细胞排列不整齐,上皮细胞增多可达4-5层,上皮深层下行纤维增多,结构致密。滑膜和肌腱均未见明显炎细胞浸润。
电针组：滑膜细胞排列基本同正常组。肌腱的外膜(腱鞘脏层)上皮细胞增生,但较模型组少,多为2-3层。上皮下结缔组织也较致密。
针刀A组：病理变化基本同电针组。
针刀B组：病理变化基本同电针组。
针刀C组：病理变化基本同电针组,但有一例可见滑膜皱襞明显增多,滑膜上皮下纤维排列较致密,在腱组织纤维内可见有的部位排列不整齐,腱细胞增多,也可见炎细胞浸润。
结论：
1肩周炎的发生与肩周炎家兔模型的活性氧自由基与抗氧化及损伤修复能力之间的平衡有着密切联系,该理论可以从多方面解释肩周炎的发病原因与病理机制.
2针刀可以提高肩周炎家兔模型清除自由基以及组织损伤修复的能力并且调节TGF-β1水平,这可能是针刀发挥作用的机制之一。
3疼痛期干预组、僵硬期干预组及疼痛期、僵硬期同时干预组的病理组织切片之间并无明显差异,这为针刀在肩周炎早期应用提供了实验依据,说明针刀在肩周炎的早期应用可以缩短病程。
Frozen shoulder, which is also called Adhesive capsulitis or painful stiff shoulder, is characterised by spontaneous onset of shoulder pain accompanied by progressive stiffness and disability. It is usually self-limiting but often has a prolonged course. is a common and frequently occurring disease for the middle-aged and elderly. At present its specific pathogenesis is unknown.
Objective
Frozen shoulder has three pathological stages, clinical studies suggest that the treatment effect of acupotomy is better in the adhesion stage. However, we believe that the therapy also can interrupt the pathological process of periarthritis, so the early use of acupotomy could shorten the course of frozen shoulder and reduce the patient suffering. In this study, the intervention effects of acupotomy were observed, through the organization of morphological and biochemical methods in order to seek the pathogenesis of frozen shoulder and the onset mechanism of acupotomy.
Methods
48 male New Zealand rabbits which were 6±0.5 months old, each weighing 2.5±0.5kg were selected. The rabbits were randomly divided into 6 groups:Normal group, model group, electroacupuncture group, acupotomy group A (pain stage of intervention), acupotomy group B (stiffness stage of intervention) only, acupotomy group C (pain and stiffness stage of joint intervention). Each group has 8 animals. Application of ice application plus strain was used to prepare the frozen shoulder rabbit models, acupotomy groups were underwent acupuncture therapy, electroacupuncture group received electroacupuncture treatment, the normal group and model group had non-intervention, and all the groups were raised under the same conditions for 33 days. The plasma, biceps, supraspinatus, and infraspinatus muscle were taken to observe the malondialdehyde levels, superoxide dismutase and total antioxidant capacity; the beginning of the biceps tendon to the far side of the tendon and the synovium were taken to observe the pathological changes by the light microscope after HE staining.
Results
1 The levels of MDA, superoxide dismutase and total antioxidant capacity in the plasma and the muscle tissue showed that the model group, the MDA level of the model group generally higher than the normal group, EA group, acupotomy group A, acupotomy group B, acupotomy group C, and had most significant differences compared with the model group; superoxide dismutase, total antioxidant capacity determination were generally lower than the normal group, EA group, acupotomy group A, acupotomy group B, acupotomy group C.
2 The pathological examination showed Normal Group:synovial:more than 1 layer of synovial cells,and some parts have seen 2 to 3 layers, folds higher in the synovial cells up to 5 to 6 layers. The deep structure of synovial cells was loose and showed few capillary. Tendon:the outer membrane of connective tissue was surrounded by the most outer epithelial cell layer of 1 to 2 cubic cells arranged in neat rows, tendon fibers were arranged regularly.
Model Group:synovial:synovial cells mostly 1 to 2 layers, more capillaries, erythrocyte sedimentation. Tendon:irregular arrangement of epithelial cells of the outer membrane, epithelial cells were increased up to 4 to 5 layers, epithelial deep down fiber increased, dense structure. Synovium and tendon were no significant infiltration of inflammatory cells.
EA group:the basic arrangement of synovial cells with normal group. Tendon of the outer membrane (tendon sheath visceral) epithelial cells, but less than the model group, mostly 2 to 3 layers. Subepithelial connective tissue was more dense.
Acupotomy group A:the same pathological changes with the basic electro-acupuncture group.
Acupotomy group B:the same pathological changes with the basic electro-acupuncture group.
Acupotomy group C:the same pathological changes with the basic electro-acupuncture group, while one case was seen significantly increased synovial plicae, synovial subepithelial fibers arranged in dense, visible in the tendon fibers arranged in irregular, some parts, tendon cells increased, but also shows infiltration of inflammatory cells.
Conclusion
1 The occurrence of the frozen shoulder in rabbit model could have closely relations to the balance between the reactive oxygen species and antioxidant, the damage and repair capacity. The theory can explain the incidence and pathological mechanisms of frozen shoulder properly
2 Acupotomy can improve the tissue injury of frozen shoulder rabbit model and free radical scavenging capacity and adjust the level of TGF-β1, which may play a role in the mechanism of Acupotomy.
3 There was no significant difference in the pathological tissue among the three intervention groups of Acupotomy, this result shows that the application of Acupotomy in the early stage of frozen shoulder can shorten the course.
研究目的：
肩周炎有明显的病理分期,目前的临床研究多认为针刀松解法对粘连期肩周炎治疗效果较好。本研究在建立家兔肩周炎模型的基础上,以针刀松解为干预手段,采用病理学和生物化学方法,从调节抗氧化能力方面寻求肩周炎的发病机制与针刀干预的起效机制,寻找针刀松解法干预肩周炎的最佳时机,为肩周炎的针刀临床治疗提供实验依据。
研究方法：
选用兔龄6±0.5个月的健康雄性新西兰家兔48只,每只体重2.5±0.5kg。随机分为6组,分别为正常组8只、模型组8只、电针组8只,针刀组A(疼痛期干预)8只、针刀组B(僵硬期干预)8只、针刀组C(疼痛期及僵硬期共同干预)8只。应用持续劳损加冰敷方法制备肩周炎家兔模型,针刀各组行针刀治疗,电针组行电针治疗,正常组、模型组,不干预,与治疗各组,同等条件下饲养,共33天。取材分别取血浆、肱二头肌、冈上肌、冈下肌、大圆肌,测定丙二醛水平、超氧化物歧化酶活力、总抗氧化能力、转化生长因子-β1水平,取肱二头肌肌腱起始处至远侧3-4mm肌腱和前下方3-4mm滑膜,固定、包埋切片,予HE染色后光镜下观察其病理改变。
实验结果：
1血浆及局部各肌肉组织的丙二醛、超氧化物歧化酶活力、总抗氧化能力的测定结果显示,模型组的丙二醛水平普遍高于正常组、电针组、针刀组A、针刀组B、针刀组C,且多数具有显著性的差异,而模型组的超氧化物歧化酶活力、总抗氧化能力测定结果要普遍低于正常组、电针组、针刀组A、针刀组B、针刀组C。
针刀组A、针刀组B、针刀组C血浆及局部各肌肉组织的抗氧化能力与脂质过氧化物水平,对比有些具有显著性差异,有些无显著性差异,但相对于模型组,以上三组的丙二醛水平相对较低,而超氧化物歧化酶活力、总抗氧化能力相对较高。
2局部病理组织观察显示,正常组：滑膜：滑膜细胞多为1层,有些部位也可见2-3层,在皱襞较多处滑膜细胞可达5-6层。滑膜细胞深层结构疏松,可见少量毛细血管。肌腱：外膜(腱鞘脏层)由结缔组织包绕,最外层上皮细胞为立方细胞1-2层,排列整齐,腱纤维排列整齐。
模型组：滑膜：滑膜细胞多为1-2层,滑膜下毛细血管较多,管腔内可见较多红细胞。肌腱：外膜上皮细胞排列不整齐,上皮细胞增多可达4-5层,上皮深层下行纤维增多,结构致密。滑膜和肌腱均未见明显炎细胞浸润。
电针组：滑膜细胞排列基本同正常组。肌腱的外膜(腱鞘脏层)上皮细胞增生,但较模型组少,多为2-3层。上皮下结缔组织也较致密。
针刀A组：病理变化基本同电针组。
针刀B组：病理变化基本同电针组。
针刀C组：病理变化基本同电针组,但有一例可见滑膜皱襞明显增多,滑膜上皮下纤维排列较致密,在腱组织纤维内可见有的部位排列不整齐,腱细胞增多,也可见炎细胞浸润。
结论：
1肩周炎的发生与肩周炎家兔模型的活性氧自由基与抗氧化及损伤修复能力之间的平衡有着密切联系,该理论可以从多方面解释肩周炎的发病原因与病理机制.
2针刀可以提高肩周炎家兔模型清除自由基以及组织损伤修复的能力并且调节TGF-β1水平,这可能是针刀发挥作用的机制之一。
3疼痛期干预组、僵硬期干预组及疼痛期、僵硬期同时干预组的病理组织切片之间并无明显差异,这为针刀在肩周炎早期应用提供了实验依据,说明针刀在肩周炎的早期应用可以缩短病程。
Frozen shoulder, which is also called Adhesive capsulitis or painful stiff shoulder, is characterised by spontaneous onset of shoulder pain accompanied by progressive stiffness and disability. It is usually self-limiting but often has a prolonged course. is a common and frequently occurring disease for the middle-aged and elderly. At present its specific pathogenesis is unknown.
Objective
Frozen shoulder has three pathological stages, clinical studies suggest that the treatment effect of acupotomy is better in the adhesion stage. However, we believe that the therapy also can interrupt the pathological process of periarthritis, so the early use of acupotomy could shorten the course of frozen shoulder and reduce the patient suffering. In this study, the intervention effects of acupotomy were observed, through the organization of morphological and biochemical methods in order to seek the pathogenesis of frozen shoulder and the onset mechanism of acupotomy.
Methods
48 male New Zealand rabbits which were 6±0.5 months old, each weighing 2.5±0.5kg were selected. The rabbits were randomly divided into 6 groups:Normal group, model group, electroacupuncture group, acupotomy group A (pain stage of intervention), acupotomy group B (stiffness stage of intervention) only, acupotomy group C (pain and stiffness stage of joint intervention). Each group has 8 animals. Application of ice application plus strain was used to prepare the frozen shoulder rabbit models, acupotomy groups were underwent acupuncture therapy, electroacupuncture group received electroacupuncture treatment, the normal group and model group had non-intervention, and all the groups were raised under the same conditions for 33 days. The plasma, biceps, supraspinatus, and infraspinatus muscle were taken to observe the malondialdehyde levels, superoxide dismutase and total antioxidant capacity; the beginning of the biceps tendon to the far side of the tendon and the synovium were taken to observe the pathological changes by the light microscope after HE staining.
Results
1 The levels of MDA, superoxide dismutase and total antioxidant capacity in the plasma and the muscle tissue showed that the model group, the MDA level of the model group generally higher than the normal group, EA group, acupotomy group A, acupotomy group B, acupotomy group C, and had most significant differences compared with the model group; superoxide dismutase, total antioxidant capacity determination were generally lower than the normal group, EA group, acupotomy group A, acupotomy group B, acupotomy group C.
2 The pathological examination showed Normal Group:synovial:more than 1 layer of synovial cells,and some parts have seen 2 to 3 layers, folds higher in the synovial cells up to 5 to 6 layers. The deep structure of synovial cells was loose and showed few capillary. Tendon:the outer membrane of connective tissue was surrounded by the most outer epithelial cell layer of 1 to 2 cubic cells arranged in neat rows, tendon fibers were arranged regularly.
Model Group:synovial:synovial cells mostly 1 to 2 layers, more capillaries, erythrocyte sedimentation. Tendon:irregular arrangement of epithelial cells of the outer membrane, epithelial cells were increased up to 4 to 5 layers, epithelial deep down fiber increased, dense structure. Synovium and tendon were no significant infiltration of inflammatory cells.
EA group:the basic arrangement of synovial cells with normal group. Tendon of the outer membrane (tendon sheath visceral) epithelial cells, but less than the model group, mostly 2 to 3 layers. Subepithelial connective tissue was more dense.
Acupotomy group A:the same pathological changes with the basic electro-acupuncture group.
Acupotomy group B:the same pathological changes with the basic electro-acupuncture group.
Acupotomy group C:the same pathological changes with the basic electro-acupuncture group, while one case was seen significantly increased synovial plicae, synovial subepithelial fibers arranged in dense, visible in the tendon fibers arranged in irregular, some parts, tendon cells increased, but also shows infiltration of inflammatory cells.
Conclusion
1 The occurrence of the frozen shoulder in rabbit model could have closely relations to the balance between the reactive oxygen species and antioxidant, the damage and repair capacity. The theory can explain the incidence and pathological mechanisms of frozen shoulder properly
2 Acupotomy can improve the tissue injury of frozen shoulder rabbit model and free radical scavenging capacity and adjust the level of TGF-β1, which may play a role in the mechanism of Acupotomy.
3 There was no significant difference in the pathological tissue among the three intervention groups of Acupotomy, this result shows that the application of Acupotomy in the early stage of frozen shoulder can shorten the course.
引文
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