针刀干预对颈椎病模型兔颈肌Beclin-1和PTEN基因和蛋白表达的影响

Effects of acupotomy therapy on expression of Beclin-1, PTEN genes and proteins in cervical muscle in cervical spondylosis rabbits

目的:观察针刀干预对颈椎病模型兔颈肌Beclin-1和PTEN基因和蛋白表达的影响,探讨针刀治疗颈椎病的可能效应机制。方法:将40只新西兰兔随机分为空白组、模型组、针刀组及电针组,每组10只,通过长期低头位造模方法建立颈椎病动物模型,造模后各组行颈椎X线侧位片,以验证造模是否成功。造模成功后针刀组行针刀干预,电针组行电针干预作对照,采用Real-time PCR法和Western blot法检测细胞自噬相关因子Beclin-1、PTEN的基因和蛋白表达水平。结果:Real-time PCR实验结果显示,模型组Beclin-1、PTEN mRNA表达水平较空白组显著升高(P<0.05,P<0.01);针刀组Beclin-1、PTEN mRNA表达水平较模型组显著降低(P<0.05);电针组Beclin-1、PTEN mRNA表达水平较针刀组显著升高(P<0.05)。Western blot实验结果显示,模型组Beclin-1、PTEN蛋白表达水平较空白组显著升高(P<0.05);针刀组Beclin-1、PTEN蛋白表达水平较模型组显著降低(P<0.05);电针组Beclin-1、PTEN蛋白表达水平较针刀组显著升高(P<0.05)。结论:针刀干预可以降低颈椎病兔颈肌Beclin-1、PTEN基因和蛋白表达水平,良性调节颈肌细胞的自噬水平,这是其治疗颈椎病的可能作用机制之一。

Objective: To observe the effects of acupotomy therapy on expression of Beclin-1, PTEN genes and proteins in cervical muscles in a rabbit cervical spondylosis model, and to study the possible mechanism of acupotomy on treatment of cervical spondylosis from the perspective of autophagy. Methods: Forty rabbits were randomly divided into four groups: control group, model group, acupotomy group and electro-acupuncture group, with 10 rabbits in each group. Cervical spondylosis model was built by prolonged flexed neck posture. After modeling, cervical X-ray lateral radiographs were performed to verify the success of modeling. After successfully modeling, acupotomy and electro-acupuncture were respectively employed in acupotomy group and electro-acupuncture group. Expression of gene and proteins of Beclin-1 and PTEN in cervical muscles were detected by Real-time PCR and Western blot. Results: The results of Real-time PCR tests showed that Beclin-1 and PTEN mRNA expression of model group increased significantly in comparison with control group(P<0.05, P<0.01). Beclin-1 and PTEN mRNA expression decreased significantly when compared acupotomy group with model group(P<0.05). Beclin-1 mRNA and PTEN mRNA expression of electro-acupuncture group significantly increased compared with acupotomy group(P<0.05). The results of Western blot tests showed that Beclin-1 and PTEN proteins expression of model group increased significantly in comparison with control group(P<0.05). Beclin-1 and PTEN proteins expression decreased significantly when compared acupotomy group with model group(P<0.05). Beclin-1 and PTEN proteins expression of electro-acupuncture group significantly increased compared with acupotomy group(P<0.05). Conclusion: Acupotomy therapy can reduce genes and proteins expression of Beclin-1 and PTEN in cervical muscles. Acupotomy therapy has a positive regulating effect on autophagy, which might be a possible mechanism of acupotomy in the treatment of cervical spondylosis.