摘要
目的:观察电针敏化穴位对结肠炎模型大鼠脑干迷走神经运动背核(DMV)乙酰胆碱转移酶阳性(Ch AT+)神经元的影响,探讨其改善大鼠结肠炎性损伤的机制。方法:将79只雄性SD大鼠随机分为正常组(20只)、正常+敏化穴位组(5只)、模型组(34只)、电针1组(15只)和电针2组(5只)。模型组和电针组均予5%葡聚糖硫酸钠(DSS)自由饮用6d建立结肠炎大鼠模型。通过尾静脉注射伊文氏蓝(EB)溶液确定模型大鼠的敏化穴位后,正常+敏化穴位组、电针1组和电针2组于敏化穴位处施以电针(疏密波,频率2 Hz/15 Hz,电流强度2 m A),每天干预30 min,电针1组连续干预6 d,正常+敏化穴位组、电针2组只干预1 d。实验第0、7、13天,评定正常组、模型组和电针1组大鼠的疾病活动指数(DAI)评分、结肠组织学损伤评分及足底机械缩足反射阈值、热痛潜伏期;实验第7天,采用免疫荧光法观察正常组、正常+敏化穴位组、模型组和电针2组大鼠脊髓背角不同板层神经元和DMV中Ch AT+神经元激活情况。结果:结肠炎模型大鼠体表EB渗出点主要分布于T12~S1节段,以L2和L5节段最集中,因此,选择L5节段的上巨虚穴作为敏化穴位。第7、13天,与正常组比较,模型组大鼠足底机械缩足反射阈值降低(P<0.001),DAI评分和结肠组织学损伤评分升高(P<0.001);第7天,模型组热痛潜伏期低于正常组(P<0.001);第13天,与模型组比较,电针1组大鼠足底机械缩足反射阈值升高(P<0.001),DAI评分和结肠组织学损伤评分降低(P<0.01,P<0.05)。与正常组比较,正常+敏化穴位组和模型组大鼠脊髓L4~L6节段背角浅层(第Ⅰ、Ⅱ板层)神经元及DMV中Ch AT+神经元激活数量增多(P<0.05,P<0.01);与正常+敏化穴位组和模型组比较,电针2组大鼠脊髓背角浅层神经元和DMV中Ch AT+神经元激活数量增多(P<0.001)。结论:结肠炎大鼠体表T12~S1节段支配区域(穴位)发生敏化;电针敏化穴位可有效缓解结肠炎模型大鼠结肠炎性损伤,其机制可能与激活脊髓背角浅层神经元和DMV中Ch AT+神经元有关。
Objective To observe the effect of electroacupuncture(EA)at sensitized acupoints on choline acetyltransferase positive(Ch AT+)neurons in dorsal motor nucleus(DMV)of brainstem vagus in the colitis model rats and explore the mechanism of the improvement in colonic inflammatory injury in the rats.Methods A total of 79 male SD rats were randomized into five groups,i.e.a normal group(20 rats),a normal plus sensitized acupoint group(5 rats),a model group(34 rats),an EA-1 group(15 rats)and an EA-2 group(5 rats).In the model group and the EA groups,5%dextran sulfate sodium(DSS)was adopted for 6-day free drinking to establish colitis model rats.By injecting Evans blue(EB)into the caudal vein in the model rats,the sensitized acupoints were determined.Afterwards,in the normal plus sensitized acupoint group,the EA-1 group and the EA-2 group,EA was exerted at the sensitized acupoints,with disperse-dense wave,2 Hz/15 Hz in frequency and 2 m A in intensity,for 30 min of intervention each day.The intervention lasted for 6 days in the EA-1 group and for 1 day in both the normal plus sensitized acupoint group and the EA-2 group.On day 0,7 and 13 of experiment,successively,the score of disease activity index(DAI),the score of colonic histological damage,as well as the changes in the mechanical paw withdrawal threshold and thermal paw withdrawal latency were evaluated in the normal group,the model group and the EA-1 group.On day 7 of experiment,using immunofluorescence staining,the activation of different lamina neurons of spinal dorsal horn and Ch AT+neurons in DMV was observed in the normal group,the normal plus sensitized acupoint group,the model group and the EA-2 group separately.Results The EB extravasating areas were distributed in the segments from T12 to S1 on the body surface of colitis model rats,mainly focusing at L2 and L5.Therefore,"Shangjuxu"(ST 37)was taken as the sensitized acupoint.Compared with the normal group on day 7 and 13 of experiment,the mechanical paw withdrawal threshold were reduced(P<0.001),DAI scores and the scores of colonic histological damage were increased(P<0.001)in the model group.Compared with the normal group on day 7 of experiment,thermal paw withdrawal latency in the model group was reduced(P<0.001).Compared with the model group on day 13 of experiment,the mechanical paw withdrawal threshold was increased(P<0.001),DAI score and the score of colonic histological damage were reduced(P<0.01,P<0.05)in the EA-1 group.Compared with the normal group,the activated numbers of the neurons in superficial laminae(ⅠandⅡ)at spinal dorsal horn of L4 to L6 and Ch AT+neurons in DMV were increased in the normal plus sensitized acupoint group and the model group(P<0.05,P<0.01).Compared with the normal plus sensitized acupoint group and the model group,the activated numbers of the neurons in superficial laminae at spinal dorsal horn of L4 to L6 and Ch AT+neurons in DMV were increased in the EA-2 group(P<0.001).Conclusion The segmental dominance(acupoints)from T12 to S1 on the body surface of colitis rats is sensitized.EA at sensitized acupoints effectively relieves colonic inflammatory injury,which is probably by activating superficial lamina neurons of spinal dorsal horn and Ch AT+neurons of DMV.
作者
于清泉
李桐
张知云
宿杨帅
何伟
王毅
张晓宁
景向红
YU Qing-quan;LI Tong;ZHNAG Zhi-yun;SU Yang-shuai;HE Wei;WANG Yi;ZHANG Xiao-ning;JING Xiang-hong(Intitute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700,China;Hubei University of TCM)
出处
《中国针灸》
CAS
CSCD
北大核心
2021年第1期45-51,共7页
Chinese Acupuncture & Moxibustion
基金
国家重点研发计划资助项目:2019YFC1709002
第四届中国科协青年人才托举工程项目:2018-2020QNRC
中国中医科学院基本科研业务费优秀青年科技人才(创新类)培养专项项目:ZZ13-YQ-069。
关键词
结肠炎
敏化穴位
迷走神经运动背核
Ch
AT+神经元
抗炎
colitis
sensitized acupoints
dorsal motor nucleus of vagus
Ch AT+neurons
anti-inflammation
