Cai Hui-Qian, Su Sheng-Yong, Zhang Xi, Qin Mei-Xiang, Huang Xiao-Zhen, Dai Qi, Jiang Xiang-Yu, Lin An, Huang Mei
The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning 530001, China.
Department of Acupuncture and Moxibustion, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023.
Zhen Ci Yan Jiu. 2020 Oct 25;45(10):799-805. doi: 10.13702/j.1000-0607.191004.
To observe the effects of mild moxibustion on the expression of autophagy and apoptosis factors Beclin-1, Bcl-2 mRNA and protein in spinal cord (including nerve root tissues) of cervical spondylotic radiculopathy (CSR) rats, so as to explore the analgesic mechanism of mild moxibustion at "Dazhui" (GV14) on CSR.
SD rats were randomly divided into blank group, model group, mild moxibustion group and mild moxibustion+3-methyladenine(3-MA) group, with 10 rats in each group. CSR model was established by inserting the wire into the cervical nerve root. The rats in the blank group were only fed normally without any intervention.The rats in the mild moxibustion group and mild moxibustion+3-MA group were given mild moxibustion at GV14 for 10 min each time,and intraperitoneal injection of 1 mL 0.9% normal saline and 1 mL 3-MA(15 mg/kg)separately. Rats in the model group were given 0.9% normal saline every day. All the three interventions were started from the 3rd day after modeling for 7 days. The rat's behavioral reaction of gait was scored and the pain threshold of rat was measured with a pain analyzer; the expressions of Beclin-1 and Bcl-2 mRNA and protein in the spinal cord (including nerve root) were detected by fluorescence quantitative PCR and immunohistochemistry, separately. The autophagosome and ultrastructure of the spinal nerve root tissue were observed by transmission electron microscope.
After modeling, the gait score was significantly increased (<0.01) and the pain threshold significantly decreased (<0.01) in the model group in comparison with the blank group. There was no statistical difference in Beclin-1 and Bcl-2 mRNA and protein expression between the blank and the model groups. After intervention, compared with the model group, the gait scores were significantly reduced (<0.01), the pain threshold and the expressions of Beclin-1 and Bcl-2 mRNA and protein were significantly increased (<0.01，<0.05) in the mild moxibustion and mild moxibustion+3-MA groups. The improvement of the above indicators as more significant in the mild moxibustion group than that in the mild moxibustion+3-MA group (<0.05). After modeling, the organelles in the spinal nerve root tissue cells of the model group were damaged and there were a small amount of autophagosomes. Compared with the model group, the ultrastructure of the spinal nerve root tissue cells in the mild moxibustion group were relatively complete, and the number of autophagosomes increased.
Mild moxibustion at GV14 has a good analgesic effect on CSR rats, which may be related to its effects in up-regulation of Beclin-1 and Bcl-2 expressions and activation of autophagy and inhibition of apoptosis.
观察温和灸对神经根型颈椎病（CSR）大鼠脊髓（含神经根组织）自噬及凋亡相关因子Beclin-1、Bcl-2 mRNA和蛋白表达的影响，以探讨温和灸“大椎”（GV14）穴治疗CSR的镇痛机制。
将SD大鼠随机分为空白组、模型组、温和灸组和温和灸+3-甲基腺嘌呤（3-MA）组，每组10只。采用将丝线插入颈神经根的方法制备CSR模型。空白组大鼠正常饲养，未作任何干预。温和灸组和温和灸+3-MA组大鼠每次给予GV14温和灸10分钟，并分别腹腔注射1 mL 0.9%生理盐水和1 mL 3-MA（15 mg/kg）。模型组大鼠每日给予0.9%生理盐水。三种干预均于造模后第3天开始，持续7天。对大鼠步态行为反应进行评分，并用疼痛分析仪测量大鼠痛阈；分别采用荧光定量PCR和免疫组化法检测脊髓（含神经根）中Beclin-1和Bcl-2 mRNA及蛋白的表达。通过透射电镜观察脊神经根组织的自噬体及超微结构。
造模后，模型组步态评分显著升高（<0.01），痛阈显著降低（<0.01），与空白组比较差异有统计学意义。空白组与模型组Beclin-1和Bcl-2 mRNA及蛋白表达差异无统计学意义。干预后，与模型组比较，温和灸组和温和灸+3-MA组步态评分显著降低（<0.01），痛阈及Beclin-1和Bcl-2 mRNA及蛋白表达显著升高（<0.01，<0.05）。上述指标温和灸组改善程度优于温和灸+3-MA组（<0.05）。造模后，模型组脊神经根组织细胞内细胞器受损，有少量自噬体。与模型组比较，温和灸组脊神经根组织细胞超微结构相对完整，自噬体数量增多。
温和灸GV14穴对CSR大鼠有良好的镇痛作用，其机制可能与上调Beclin-1和Bcl-2表达、激活自噬及抑制凋亡有关。
