世界中医药

目的:观察电针对颈椎病大鼠椎间盘软骨细胞及MCP-1/CCR2信号通路的影响。方法:将45只雄性SD大鼠,按照随机数字表法分为3组,假手术组、模型组和电针组,每组15只。除假手术组外均建立动静力失衡性颈椎病模型,并于造模后3个月开始电针干预双侧颈夹脊穴,30 min/次,1次/d,14 d为1个疗程。研究中通过电子透射电镜观察颈椎间盘软骨细胞的形态判定颈椎病造模是否成功;而后采用免疫组化检测颈椎间盘软骨细胞中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达情况;接着采用实时聚合酶链式反应(Real time PCR)检测MCP-1/CCR2信号通路,观察电针对MCP-1/CCR2信号通路的影响。结果:1)假手术组的颈椎间盘软骨细胞表面较光滑,可见微绒毛样凸起,细胞整体形态上类似椭圆形,细胞核形态类似一个肾脏,且细胞核完整无破损,染色质均匀;而细胞外的基质中含有丰富且排列有序的胶原纤维;模型组中的颈椎间盘软骨细胞出现退行性改变,其细胞表面的微绒毛样凸起明显减少,细胞核破损,染色质明显固缩,细胞外基质中的胶原纤维明显减少,排列无序。提示颈椎病造模成功。2)与假手术组比较,模型组和电针组大鼠TNF-α和IL-6表达均较密集,阳性表达增多(P<005),其中是电针组TNF-α和IL-6表达明显较模型组降低(P<005),提示电针能够抑制颈椎病大鼠椎间盘软骨细胞的TNF-α和IL-6表达。3)与假手术组比较,模型组和电针组颈椎间盘中MCP-1/GAPDH和CCR2/GAPDH均增高,差异有统计学意义(P<005);电针组与模型组比较,颈椎间盘中MCP-1/GAPDH和CCR2/GAPDH低于模型组,差异有统计学意义(P<005)。结论：电针可以抑制颈椎病大鼠椎间盘软骨细胞中TNF-α和IL-6表达,可能与抑制MCP-1/CCR2信号通路的激活有关。

English Summary:

To observe the effect of electroacupuncture on chondrocyte and MCP-1/CCR2 signaling pathway of intervertebral disc in rats with cervical spondylosis.Methods:A total of 45 male SD rats were divided into three groups according to random number table，a sham operation group，a model group and an electro-acupuncture group，with 15 rats in each group.Except for the sham operation group，the models of dynamic and static imbalance cervical spondylosis were established，and electro-acupuncture intervention on bilateral cervical Jiaji(EX-B2)acupoints was started 3 months after modeling，30 minutes a day for 14 days.In this study，the morphology of cervical intervertebral disc chondrocyte was observed by transmission electron microscopy to determine the success of cervical spondylosis modeling.Then the expression of tumor necrosis factor-alpha(TNF-alpha)and interleukin-6(IL-6)in cervical intervertebral disc chondrocyte was detected by immunohistochemistry.Then real-time polymerase chain reaction(Real-time PCR)was used to detect MCP-1/CCR2 signaling pathway and observe the effect of electroacupuncture on MCP-1/CCR2 signaling pathway.Results:1)The surface of cervical intervertebral disc chondrocyte in sham operation group was smooth，with microvilli-like protuberance，oval shape of cell as a whole，and the shape of nucleus was similar to that of a kidney，and the nucleus was intact without damage and the chromatin was uniform; the extracellular matrix contained abundant and orderly collagen fibers; the degeneration of cervical intervertebral disc chondrocyte appeared in the model group.The microvilli-like protrusions on the cell surface were significantly reduced，the nucleus was damaged，the chromatin was obviously condensed，and the collagen fibers in the extracellular matrix were significantly reduced and arranged in disorder.It is suggested that cervical spondylosis was successfully modeled.2)Compared with sham operation group，the expression of TNF-alpha and IL-6 in model group and electro-acupuncture group were more intensive and positive(P<005).Among them，the expression of TNF-alpha and IL-6 in electro-acupuncture group was significantly decreased than that in the model group(P<005)，suggesting that electro-acupuncture could inhibit the expression of TNF-alpha and IL-6 in intervertebral disc chondrocytes of rats with cervical spondylosis.3)Compared with sham operation group，MCP-1/GAPDH and CCR2/GAPDH in cervical intervertebral disc of the model group and the electro-acupuncture group increased，with statistical significance(P<005); compared with model group，MCP-1/GAPDH and CCR2/GAPDH in cervical intervertebral disc of electro-acupuncture group were lower than model group，with statistical significance(P<005).Conclusion:Electroacupuncture can inhibit the expression of TNF-alpha and IL-6 in the chondrocyte of intervertebral disc of rats with cervical spondylosis，which may be related to the inhibition of the activation of MCP-1/CCR2 signaling pathway.

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