项七针对颈椎间盘退变大鼠颈椎间盘低氧诱导因子1α及Notch1表达的影响

Effect of“nape seven needles”on expression of hypoxia inducible factor-1αand Notch1 in cervical intervertebral disc degeneration rats

目的:观察项七针对颈椎间盘退变(CDD)大鼠颈椎间盘中低氧诱导因子1α(HIF-1α)及Notch1表达的影响,探讨项七针治疗CDD的分子机制。方法:SD大鼠随机分为正常组、模型组、非穴组及项七针组,每组10只。动静力失衡法制备CDD大鼠模型。项七针组选取“风府”和双侧“天柱”“风池”“完骨”进行针刺,非穴组针刺尾尖及腋下非经非穴,两组均留针20 min,每日治疗1次,治疗6 d,休息1 d,共治疗4周。采用斜板实验评估各组大鼠的运动功能;螺旋CT评估颈椎退变情况;Western blot法、免疫组织化学法及实时荧光定量PCR法检测颈椎间盘组织中HIF-1α和Notch1蛋白及mRNA的表达水平。结果:治疗后,与正常组比较,模型组大鼠斜板角度减小(P<0.05),CT评分、颈椎间盘组织中HIF-1αmRNA表达水平均升高(P<0.05);与模型组和非穴组比较,项七针组大鼠斜板角度明显增大(P<0.05),CT评分降低(P<0.05),颈椎间盘中HIF-1α、Notch1阳性表达增加(P<0.05),HIF-1αmRNA、Notch1蛋白及mRNA表达水平均升高(P<0.05)。结论:项七针治疗可以减轻大鼠CDD程度,可能与上调大鼠颈椎间盘组织中HIF-1α和Notch1的表达有关。

Objective To observe the effect of“nape seven needles”on the expressions of hypoxia inducible factor-1α(HIF-1α)and Notch1 in cervical intervertebral disc of cervical disc degeneration(CDD)rats,so as to reveal its underlying mecha-nisms in improving CDD.Methods SD male rats were randomly divided into normal,model,non-acupoint and nape seven needles groups,with 10 rats in each group.Staticdynamic imbalance method was used to establish CDD model.Rats in the nape se-ven needles group were treated with acupuncture at“Fengfu”(GV16),and bilateral“Fengchi”(GB20),“Wangu”(GB12)and“Tianzhu”(BL10),and rats in the non-acupoint group received acupuncture at the sham acupoints at the caudal tip and armpit,both for 20 min,6 days a week for 4 weeks.After intervention,tilted plane test and spiral CT were used to assess the neck movement function and cervical degeneration degree of rats;immunohistochemistry was used to observe the protein expression levels of HIF-1αand Notch1 in cervical discs tissue;Western blot and real-time quantitative PCR were used to detect the protein and mRNA expression levels of HIF-1αand Notch1 in cervical discs tissue,respectively.Results After modeling,the cervical curvature was straightened,with narrowed intervertebral space,rough and hardened articular surface,osteophytes,and blurred articular space and articular process,which was relatively milder in the nape seven needles group.Compared with the normal group,the angle of tilted plane was significantly reduced(P<0.05),while cervical scores,HIF-1αmRNA expression level in cervical intervertebral disc tissue were significantly increased(P<0.05)in the model group.Compared with the model and the non-acupoint groups,cervical scores were significantly reduced(P<0.05),while the angle of tilted plane,HIF-1αand Notch1 positive expressions,HIF-1αmRNA expression level,Notch1 protein and mRNA expression levels in cervical intervertebral disc tissue were significantly increased(P<0.05)in the nape seven needles group.Conclusion Acupuncture of“nape seven needles”can reduce the degree of cervical degeneration in rats,which was possibly associated with its effects in up-regulating the expressions of HIF-1αand Notch1 in cervical intervertebral disc tissue,promoting the proliferation and recovery of endogenous cells in nucleus pulposus.