针刀干预颈椎间盘退变模型兔的实验研究

发布时间：2018-03-05 10:33

本文选题：针刀　切入点：针刺　出处：《南京中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的：通过“低头位+风寒湿刺激”的方法建立颈椎间盘退变动物模型,用针刀干预模型动物,从组织形态学及椎间盘细胞外基质组分、代谢相关酶和细胞因子含量变化等方面来探讨针刀疗法能否通过延缓甚至修复椎间盘退变来达到治疗颈椎病的目的。方法：购进6+0.5月龄健康雄性新西兰大白兔,将其分为空白组、模型组、针刀组和针刺组,每组6只。除对照组外其余各组均用“低头位+风寒湿刺激”的方法建立实验模型。造模结束后,通过对比空白组兔和造模组兔颈椎X线,初步判定造模是否成功。造模成功后,模型组常规饲养,用针刀和毫针分别对针刀组和针刺组进行干预治疗,针刀组每周干预1次,共干预4次,针刺组每周干预3次,共干预12次。干预结束后,HE染色,光镜下观察四组实验动物颈部肌肉及椎间盘的组织形态学变化。用ELISA法检测四组实验动物椎间盘内MMP-3、TGF-β1含量,Western blot法检测椎间盘内II型胶原、蛋白多糖的蛋白表达水平。结果：空白组动物颈椎X线显示：颈椎生理曲度正常存在,椎间隙无增宽或狭窄,椎体排列整齐,椎体周围光滑无骨赘形成。造模后的实验动物颈椎X线显示：颈椎正常生理曲度消失,变直甚至反弓,椎间隙变窄,椎体排列不整齐,椎体边缘毛糙有骨赘形成。针刀及针刺干预结束后,颈部肌肉HE染色光镜下观察显示：空白组实验动物颈肌组织切片可见肌肉组织结构清晰,肌细胞无变性水肿,无炎性细胞浸润。模型组可见肌纤维排列不齐、断裂。肌细胞肿胀变形,肌核分布不均,大量炎性细胞浸润。针刀组可见肌肉组织结构较模型组整齐,炎性细胞浸润明显减少,细胞核排列较前均匀整齐。针刺组可见肌纤维排列较模型组明显整齐,肌细胞仍有轻度肿胀、扭曲,间质内有少量炎性细胞浸润,但较模型组明显减轻。椎间盘HE染色光镜下观察显示：空白组髓核居中,大小正常,纤维环包绕四周,纤维排列整齐。模型组可见髓核明显固缩,纤维排列杂乱、扭曲变形,甚至断裂。针刀组、针刺组均仍可见髓核固缩,但较之模型组,明显改善。周围纤维排列较模型组明显整齐,裂隙变小。ELISA结果：模型组兔椎间盘内MMP-3含量较空白组明显升高,二者有极明显差异(P0.01)；针刺组与模型组相比明显降低,有明显差异(P0.05)；针刀组与模型组相比明显降低,有极明显差异(P0.01)；针刀组与针刺组偏低,有极显著差异(P0.01)。模型组兔椎间盘内TGF-β1的含量较空白组明显降低,二者有极明显差异(P0.01)；针刺组与模型组相比明显升高,有明显差异(P0.05)；针刀组与模型组相比明显升高,有极明显差异(P0.01)；针刀组比针刺组略升高,无统计学意义(P0.05)。Western blot结果：所测四组实验动物椎间盘中Ⅱ型胶原蛋白表达水平模型组较空白组明显降低,二者有极明显差异(P0.01)；针刺组与模型组相比明显升高,有明显差异(P0.05)：针刀组与模型组相比明显升高,有极明显差异(P0.01)；针刀组较针刺组偏高,有显著差异(P0.05)。所测四组实验动物椎间盘中蛋白多糖蛋白表达水平较空白组明显降低,二者有极明显差异(P0.01)；针刺组与模型组相比明显升高(P0.01)；针刀组与模型组相比明显升高,有明显差异(P0.05)；针刺组较针刀组略偏高,但无统计学意义(P0.05)。结论：1.对比空白组和模型组颈椎X线、颈肌及椎间盘组织形态学变化,“低头位+风寒湿”刺激可成功建立椎间盘退变实验模型,该实验模型是成功的。2.针刀及针刺干预均能使椎间盘中蛋白多糖和II型胶原蛋白表达升高,使MMP-3含量降低、TGF-β1含量升高,说明针刀及针刺干预治疗颈椎病可能是通过对椎间盘内细胞因子及相关代谢酶的良性调节,抑制细胞外基质的降解,缓解甚至修复椎间盘的退变。3.从颈肌、椎间盘组织形态及椎间盘微量物质的对比,总体来讲,针刀疗效略优于针刺。
[Abstract]:Objective: to establish the model of cervical intervertebral disc degeneration animal by means of "bow + cold wet stimulation, the animal model with Acupotomy intervention from histomorphology and disc extracellular matrix components, metabolism related enzymes and cytokines content, to explore whether by acupotomy therapy can delay or even repair of intervertebral disc degeneration to achieve the treatment of cervical spondylosis. Methods: the purchase of 6+0.5 month old male New Zealand rabbits were divided into blank group, model group, acupuncture group and acupuncture group, 6 rats in each group. All groups except the control group by method of" head + wind cold stimulation of the experimental model is established after the modeling. By contrast, the blank group and model group rabbit rabbit cervical X-ray, determine the initial success of modeling. After successful modeling, the model group, with the needle knife and needle of acupotomy group and acupuncture group intervention, Acupotomy group Intervention 1 times a week, a total of 4 times a week group of acupuncture intervention, intervention 3 times, a total of 12 times. The intervention after the intervention, HE staining, the morphological changes of muscle and disc four groups of experimental animal neck under light microscope. ELISA was used to detect four groups of experimental animal intervertebral disc MMP-3, TGF- beta 1 content II Western, the detection of intervertebral disc by blot collagen, the expression level of protein polysaccharide protein. Results: blank group: Animal cervical X-ray normal cervical lordosis, intervertebral broadening or stenosis, vertebral body arranged around the vertebral body smooth without osteophyte formation. Experimental animal model: cervical X-ray disappeared the normal cervical lordosis, straight and even anti bow, narrowing of intervertebral space, vertebral malalignment, rough edge vertebral osteophyte formation. Acupuncture and acupuncture intervention after the neck muscle HE staining light microscope observation showed that: blank group experimental animal muscle Histological sections showed the muscle tissue structure is clear, muscle cells without edema, no inflammatory cell infiltration. The model group showed that the muscle fibers were uneven fracture. Muscle cell swelling, muscle nuclear uneven distribution, a large number of inflammatory cells infiltration. Acupotomy group visible muscle tissue structure orderly compared with model group, inflammatory cell infiltration reduce nuclear arrangement was uniform. The acupuncture group showed muscle fibers were compared with the model group was neat, muscle cells still had mild swelling, distortion, interstitial infiltration of inflammatory cells, but was significantly reduced compared with model group. The intervertebral disc HE staining by light microscope observation showed that: blank group nucleus center size normally, fiber ring wrapped around arrangedregularly. Model group showed obvious nucleus pyknosis, fibers arranged disorderly, distorted, or even fracture. Acuknife group, the acupuncture group was still visible nucleus pyknosis, but significantly compared with the model group, improve the week. Wai fibers were arranged in the model group was significantly smaller in order, crack.ELISA results: in the model group, the rabbit intervertebral disc MMP-3 was significantly higher than that in blank group two there were significant differences (P0.01); acupuncture group compared with model group significantly decreased, had significant difference (P0.05); the knife needle group and the model group significantly decreased there were significant differences (P0.01); low Acupotomy group and acupuncture group, there were significant differences between the model group (P0.01). The content of the rabbit intervertebral disc TGF- beta 1 was significantly lower than those in the control group two, there were significant differences (P0.01); Acupuncture group compared with model group significantly increased, there was significant difference (P0.05); acupuncture group compared with model group increased significantly (P0.01); acupuncture group than in acupuncture group increased slightly, no statistical significance (P0.05).Western blot results: the measured four groups of experimental animal intervertebral discs of type II collagen expression in model group than in the blank group Significantly, the two have very significant difference (P0.01); acupuncture group compared with model group increased significantly, there was significant difference (P0.05): acupuncture group compared with model group increased significantly (P0.01); Acupotomy group than in the acupuncture group, there was significant difference (P0.05). The four group the experimental animal intervertebral disc proteoglycan expression levels were significantly lower than those in the control group two, there were significant differences (P0.01); acupuncture group and model group were significantly higher than (P0.01); acupuncture group compared with model group increased significantly and had significant difference (P0.05); acupuncture group compared with Acupotomy group is slightly higher, but no statistical significance (P0.05). Conclusion: 1. compared with the blank group and the model group of cervical intervertebral disc and cervical X-ray, histological changes, "bow + cold wet" stimulation of intervertebral disc degeneration can be successfully established the experimental model, the experimental model is.2. and the needle acupuncture intervention can work The intervertebral disc proteoglycan and type II collagen expression increased, and decreased MMP-3 contents, increased TGF- beta 1 content, indicating needle knife and acupuncture intervention for the treatment of cervical spondylosis may be through regulating on intradiscal cytokines and related metabolic enzymes, inhibition of extracellular matrix degradation, alleviate or even repair the degeneration of intervertebral disc.3. from the neck, contrast, intervertebral discs and disc trace substances in general, Acupotomy is better than acupuncture.

【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245;R-332

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