电针对神经根型颈椎病大鼠神经损伤的胶原纤维、TGF-β、TNF-α的影响

Effects of Electroacupuncture on Collagen Fibers,TGF-β and TNF-α Induced by Nerve Injury in Rats with Radiculotic Cervical Spondylosis

目的研究电针干预神经根型颈椎病神经损伤的作用机制。方法选用72只雌性Wistar大鼠,随机分为空白对照组,假手术组,模型组,电针组,每组18只;建立神经根型颈椎病模型后,造模第3 d开始电针刺激大鼠的双侧的天柱、颈百劳、大杼穴,隔天1次,每次20 min;造模后第4周,检测大鼠的痛阈后并取颈部脊髓组织;石蜡切片,Masson染色脊髓组织检测胶原纤维;免疫组化检测颈部脊髓组织的TGF-β、TNF-α的蛋白表达水平;qPCR检测脊髓组织中TGF-β、TNF-α的mRNA表达水平。结果①痛阈检测结果:与空白组比较,假手术组的差异不具备统计学意义(P>0.05);模型组与假手术组比较,大鼠的痛阈值明显下降,电针组与模型组比较,大鼠痛阈值显著提高(P<0.05)。②Masson染色后,与空白组对照组相比,假手术组胶原纤维未见明显变化(P>0.05);与假手术组相比,模型组胶原纤维明显增加(P<0.05);与模型组相比,电针组胶原纤维明显减少(P<0.05)。③免疫组化检测结果,与空白对照组比较,假手术组TGF-β、TNF-α蛋白未见明显变化(P>0.05);与假手术组相比,模型组TGF-β、TNF-α蛋白明显增加(P<0.05);与模型组相比,电针组TGF-β、TNF-α蛋白明显减少(P<0.05)。④qPCR检测结果,空白对照组与假手术组相比,差异没有统计学意义(P>0.05);与假手术组相比,模型组TNF-αmRNA、TGF-β1mRNA表达水平均显著上升(P<0.05);与模型组相比,电针组TNF-αmRNA、TGF-β1mRNA表达水平明显降低(P<0.05)。结论电针治疗神经根型颈椎病大鼠模型,通过下调颈髓组织中TGF-β、TNF-α蛋白及mRNA表达水平,提高神经根型颈椎病大鼠的痛阈值并降低胶原纤维的含量,减少神经外瘢痕组织的增生,从而减轻神经的炎性损伤及促进神经损伤的修复。

Objective:To study the mechanism of electrioacupuncture intervention on nerve injury in cervical radiculopathy.Methods:72 female Wistar rats were randomly divided into blank control group,sham operation group,model group and electroacupuncture group,18 rats each group.After establishing the cervical spondylosis model of nerve root type,on the 3rd day of modeling,the rats'bilateral Tianzhu,Jingbailao and Dashu points were stimulated by electroacupuncture,once every other day for 20 min each time.At the 4th week after modeling,the rats'pain threshold was detected and their cervical spinal cord tissues were taken.Paraffin sections were obtained,and collagen fibers were detected by Masson staining of spinal cord.The protein expression levels of TGF-βand TNF-αwere detected by immunohistochemistry.The mRNA expression levels of TGF-βand TNF-αin the spinal cord were detected by qPCR.Results:1.The results of pain threshold detection:Compared with the blank group,the difference of the sham operation group was not statistically significant(P>0.05).Compared with that of the sham operation group,the pain threshold of rats of the model group was significantly decreased,and the pain threshold of rats of the electroacupuncture group was significantly increased(P<0.05).2.After Masson staining,there was no significant change in collagen fibers of the sham operation group compared with that of the blank control group(P>0.05);Compared with that of the sham operation group,the collagen fibers of the model group were significantly increased(P<0.05).Compared with that the model group,the collagen fibers of the electroacupuncture group were significantly reduced(P<0.05).3.Compared with that of the blank control group,the TGF-βand TNF-αproteins of the sham operation group had no significant changes(P>0.05).Compared with that of the sham operation group,the TGF-βand TNF-αproteins of the model group were significantly increased(P<0.05).Compared with that of the model group,the TGF-βand TNF-αproteins of the electroacupuncture group were significantly decreased(P<0.05).④There was no significant difference in qPCR results between the blank control group and the sham operation group(P>0.05).Compared with that of the sham operation group,the expression levels of TNF-αmRNA and TGF-β1mRNA of the model group were significantly increased(P<0.05).Compared with that of the model group,the expression levels of TNF-αmRNA and TGF-β1mRNA of the electroacupuncture group were significantly decreased(P<0.05).Conclusion:Electroacupuncture can reduce the expression levels of TGF-βand TNF-αprotein and mRNA in cervical spinal cord tissue,increase the pain threshold of CSR rats,reduce the content of collagen fibers,and reduce the hyperplasia of extraneuronal scar tissue,so as to alleviate the inflammatory injury of nerves and promote the repair of nerve injury.