电针联合壮医药线点灸对带状疱疹后遗神经痛模型大鼠脑组织NR1、NR2B的影响

Effect of Electroacupuncture Combined with Zhuang Medicine Medicated Thread Moxibustion on NR1 and NR2B in Posterior Herpes Zoster Neuralgia Rats

目的观察电针、药线点灸、电针合药线点灸对带状疱疹后遗神经痛模型大鼠的镇痛效果及作用机制。方法将50只大鼠随机等分为5组,10只为空白对照组,另40只采用RTX腹腔注射来复制实验模型,造模成功后随机等分为模型对照组、电针治疗组、药线点灸治疗组、电针合药线点灸治疗组,共干预14 d。观察各组机械性痛阈(Paw withdrawl threshold,PWT)的改变,取脑组织,采用免疫印迹法(Western blot,WB)法检测NR1、NR2B磷酸化水平的变化,采用实时荧光定量检测(real-time fluorescencespectrometry,QPCR)NR1、NR2B mRNA表达的变化。结果(1)PWT:在注射后1、4、7、14、21、36 d检测,模型对照组的大鼠PWT值依次降低,与空白对照组对比有显著差异(P<0.001);与模型对照组比较,3个治疗组PWT值均显著提高(P<0.001),其中药线点灸治疗组较电针治疗组提高更为明显(P<0.01),电针合药线点灸治疗组提高最为明显(P<0.001)。(2)WB结果:采用IPP软件蛋白条带灰度值进行分析,结果表明,与空白组比较,模型组NR1,P-NR1、NR2B、P-NR2B的表达明显增加(P<0.05);与模型组比较,电针治疗组,药线点灸治疗组和电针合药线点灸治疗组,P-NR1及P-NR2B蛋白表达量均明显降低(P<0.05)。(3)NR1 mRNA表达:模型对照组脑组织中NR1 mRNA表达水平最高,明显高于其余4组,数据差异有统计学意义(P<0.001);各治疗组组间对比,电针和药线点灸组的NR1 mRNA表达水平明显低于单纯电针治疗组和药线点灸治疗组,差异显著(P<0.001)。(4)NR2B mRNA表达:模型对照组、电针治疗组、药线点灸治疗组NR2B mRNA表达量较空白对照组均明显升高(P<0.001),其中模型对照组的NR2B mRNA表达量显著高于电针治疗组、药线点灸治疗组和电针合药线点灸治疗组(P<0.01);各治疗组中电针合药线点灸治疗组的NR2B mRNA表达量最低。结论电针及药线点灸对大鼠的机械性痛阈具有显著的提升效果,可减轻大鼠后遗神经痛,两种疗法联合治疗可起到协同增效的作用,其镇痛机制可能与下调脑组织中NR1、NR2B水平相关。

Objective To observe the analgesic effects and its mechanisms of electroacupuncture(EA),medicated thread moxi⁃bustion and the combination of EA and medicated thread moxibustion on postherpetic neuralgia model rats.Methods Fifty rats were randomly and equally divided into five groups,10 of which were blank control group,and the other 40 rats were treated with intraperitoneal injection of RTX as the experimental model.After successful modeling,they were randomly and equally divided in⁃to the model control group,EA treatment group,medicated thread moxibustion treatment group and EA combined with medicated thread moxibustion treatment group,for a total of 14 days.The changes of Paw withdrawl threshold(PWT)in each group were ob⁃served.The brain tissue was taken out.The phosphorylation levels of NR1 and NR2B were detected by western blot(WB)meth⁃od,and the mRNA expression levels of NR1 and NR2B were quantified by real-time fluorescence spectrometry(QPCR).Results(1)PWT:PWT values of rats in the model control group were successively decreased and significantly different from those of the blank control group detected on day 1,4,7,14,21 and 36 after injection(P<0.001).Compared with those of the model control group,the PWT values of the three treatment groups were significantly increased(P<0.001),and the improvement in the medi⁃cated thread moxibustion treatment group was more significant than that in the EA treatment group(P<0.01).The improvement in the EA combined with medicated thread moxibustion treatment group was mostly significant(P<0.001).(2)WB results:thegray values of protein bands in IPP software were analyzed.The results showed that compared with those of the blank group,the expressions of NR1,P-NR1,NR2B and P-NR2B in the model group were significantly increased(P<0.05).Compared with those of the model group,the protein expression levels of P-NR1 and P-NR2B in the EA treatment group,the medicated thread moxibustion treatment group and the EA combined with medicated thread moxibustion treatment group were significantly reduced(P<0.05).(3)NR1 mRNA expression:the expression level of NR1 mRNA in the brain tissue of the model control group was the highest,significantly higher than that of the other four groups,and the difference between the data was statistically significant(P<0.001).As for the inter-group comparison,the NR1 mRNA expression levels in the EA and medicated thread moxibustion groups were significantly lower than those in the EA treatment group and the medicated thread moxibustion group,with significant differences(P<0.001).(4)NR2B mRNA expression:NR2B mRNA expression levels in the model control group,the EA treat⁃ment group and the medicated thread moxibustion treatment group were significantly higher than that in the blank control group(P<0.001),and NR2B mRNA expression level in the model control group was significantly higher than that in the EA treatment group,the medicated thread moxibustiontreatment group and the EA combined with herbal line moxibustion treatment group(P<0.01).Among all the treatment groups,the NR2B mRNA expression in the EA and medicated thread moxibustion treatment group was the lowest.Conclusion Both EA and medicated thread moxibustion have significant effect on increasing the mechanical pain threshold of rats and reducing the postherpetic neuralgia.The combination of the two therapies can play a synergistic role,and its analgesic mechanism may be related to the down-regulation of NR1 and NR2B levels in the brain tissue.