[Visual acupotomy intervention mitigates pain reaction by improving intervertebral disc degeneration and inhibiting apoptosis of nucleus pulposus cells in rabbits with cervical spondylosis].

To investigate the effect of visual acupotomy intervention on intervertebral disc degeneration, nucleus pulposus cell apoptosis and expression of apoptosis related proteins in rabbits with cervical spondylosis (CS), so as to explore its mechanism underlying improvement of CS.A total of 48 male New Zealand rabbits were randomly divided into blank control, model, acupotomy and medication (meloxicam) groups, with 12 rabbits in each group. The neck type CS model was established by forcing the rabbit to make a neck flexion for 5 hours in a restrained chamber, once daily for 12 weeks. Rabbits of the medication group received an intramuscular injection of meloxicam (0.35 mg/kg), once daily for 4 consecutive weeks, and those of the acupotomy group received ultrasound-guided acupotomy intervention, once a week for 4 weeks. The pain threshold (PT) was measured by using a VonFrey electronic pain detector. The levels of prostaglandin E2 (PGE2), 5-hydroxytryptamine (5-HT) and substance P (SP) in serum were detected by ELISA. The severity of intervertebral disc degeneration was observed by using magnetic resonance imaging (MRI) and given scores in accordance with Suzuki's and colleague's "new classification system of cervical disk degeneration". The apoptosis of nucleus pulposus cells was analyzed by TUNEL staining. The protein expression levels of apoptosis-related protein Fas, cysteinyl aspartate-specific protease-3 (Caspase-3), B-cell lymphoma-2 asso-ciated X protein (Bax) and B-cell lymphoma-2 protein (Bcl-2) were measured by Western blot.Compared with the blank control group, the PT and Bcl-2 expression and MRI score were significantly down-regulated (P<0.01, P<0.001), whereas the contents of serum PGE2, 5-HT and SP, ratios of TUNEL-positive cells, and expression of Fas, Caspase-3 and Bax were considerably up-regulated (P<0.001, P<0.05, P<0.01) in the model group. In contrast to the model group, both the medication and acupotomy groups had an obvious increase in the levels of PT and Bcl-2 expression and MRI score (P<0.05, P<0.01), and a significant decrease in the contents of serum PGE2, 5-HT, SP, ratios of TUNEL-positive cells, and expression of Fas, Caspase-3 and Bax proteins (P<0.05). No significant differences were found between the medication and acupotomy groups in all the indexes mentioned above (P>0.05).Visual acupotomy intervention can mitigate the pain state of CS rabbits, which may be related to its functions in improving the intervertebral disc degeneration, reducing inflammatory reactions and apoptosis of nucleus pulposus cells.目的：观察可视化针刀对颈型颈椎病（CS）兔椎间盘退变、髓核细胞凋亡和凋亡相关蛋白表达的影响，探讨针刀治疗CS的可能机制。方法：将新西兰兔随机分为空白对照组、模型组、美洛昔康组和针刀组，每组12只。采用改良长期低头位法制备颈型CS兔模型。美洛昔康组予美洛昔康（0.35 mg/kg）肌肉注射，每日1次，连续4周；针刀组予超声引导下针刀干预，每周1次，连续4周。应用VonFrey电子痛阈仪测定新西兰兔压力痛阈值；ELISA法检测血清中前列腺素E2（PGE2）、5-羟色胺（5-HT）和P物质（SP）的含量；MRI观察新西兰兔颈椎间盘并以Suzuki分级法评价其退变程度；TUNEL染色法观察颈椎间盘髓核细胞凋亡情况并计算凋亡率；Western blot法检测颈椎间盘髓核凋亡相关因子Fas、半胱氨酸天冬氨酸特异性蛋白酶3（Caspase-3）、B淋巴细胞瘤-2（Bcl-2）及其相关的X蛋白（Bax）蛋白表达量。结果：与空白对照组比较，治疗前模型组、美洛昔康组、针刀组和治疗后模型组压力痛阈值均显著降低（P<0.01）；与模型组比较，治疗后美洛昔康组和针刀组压力痛阈值显著升高（P<0.05）。与空白对照组比较，模型组血清中PGE2、5-HT、SP含量显著升高（P<0.001，P<0.05），颈椎间盘信号显著降低（P<0.001），颈椎间盘髓核细胞凋亡率及Fas、Caspase-3、Bax蛋白表达量显著升高（P<0.01，P<0.001），Bcl-2蛋白表达量显著降低（P<0.01）；与模型组比较，美洛昔康组和针刀组血清中PGE2、5-HT、SP含量显著降低（P<0.05），椎间盘信号显著增强（P<0.05, P<0.01），髓核细胞凋亡率及Fas、Caspase-3、Bax蛋白表达量显著降低（P<0.05），Bcl-2蛋白表达量显著升高（P<0.05）。结论：可视化针刀可抑制髓核细胞凋亡，从而改善CS兔的颈椎间盘退变，这可能是针刀治疗CS的作用机制之一。.