穴位埋线治疗变应性鼻炎的神经免疫联动机制研究  

Effects of Catgut Implantation at Acupoints on Neural Immune Linkage of Rats with Allergic Rhinitis

作　　者：杨莎莎[1] 曾斌[1] 刘代恩 李昕蓉[2] 刘洋[2] 向南[1] 袁淑贤[1] 张勤修[2] 

机构地区：[1]成都中医药大学,四川成都610072 [2]成都中医药大学附属医院耳鼻咽喉科,四川成都610072

出　　处：《中华中医药学刊》2017年第10期2480-2484,I0002-I0006,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81273985;81473523);国家"十二五"科技支撑计划项目(2015BA I04B00)

摘　　要：目的:探讨穴位埋线治疗变应性鼻炎(AR)的可能作用机制一神经免疫联动机制,即穴位埋线通过下调神经肽P物质(SP),进而对肥大细胞膜上游TLR2、TLR4及下游NF-κBp65信号转录调节通路产生影响,最终抑制肥大细胞脱颗粒。方法:48只SD大鼠随机分为6组:正常对照组,模型组,布地奈德治疗组,穴位埋线治疗1周、2周、3周组。除正常对照组外,其余各组均采用卵清蛋白致敏SD大鼠的方法制作变应性鼻炎动物模型。造模成功后,进行穴位埋线的3组均在迎香穴(LI20)、曲池穴(LI11)、足三里穴(ST36)埋入2-0羊肠线,分别经治疗1、2、3周阶段性观察,布地奈德组给予布地奈德滴鼻剂5μg/(kg·d)滴鼻,每日1次,模型组维持卵清蛋白滴鼻,正常组给予生理盐水滴鼻。观察内容:(1)大鼠变应性鼻炎相关的行为学评分变化;(2)HE染色,观察各组鼻黏膜病理形态学改变;(3)免疫组织化学法分别检测各组鼻黏膜组织中感觉神经肽(SP)、NF-κBp65蛋白的表达。(4)免疫荧光双染法观察TLR2、TLR4在肥大细胞膜表面的蛋白表达差异。(5)甲苯胺蓝染色法检测各组鼻黏膜组织中肥大细胞脱颗粒率的差异。结果:随穴位埋线周期的延长,埋线第3周较第1、2周、布地奈德治疗组可明显改善AR症状,具有统计学意义(P<0.01)。HE染色显示埋线治疗组、布地奈德组大鼠鼻黏膜纤毛倒伏、脱落及嗜酸性粒细胞浸润等状况较埋线模型组减轻。免疫组织化学检测:与模型组相比,埋线第3周大鼠鼻黏膜组织中SP、NF-κBp65蛋白含量降低,具有统计学意义(P<0.05)。免疫荧光双标检测:与模型组相比,布地奈德治疗组、埋线治疗第3周TLR2平均光密度降低,具有统计学意义(P<0.05);布地奈德治疗组TLR4平均光密度明显降低,具有非常显著的统计学意义(P<0.01);其余各组不具备统计学意义。甲苯胺蓝染色检测:鼻黏膜组织中肥大细胞脱颗粒率,与模型组相比,穴位�Objective ： To explain the mechanisms on neural immune linkage of rats with allergic rhinitis treated by catgut implantation at acupoints by cutting neuropeptide substance P（SP） and then mast cell membrane upstreams TLR2 and TLR4 and downstreams NF - κBp65 transcriptional regulation signal pathway eventually to inhibit mast cell degranulation. Methods：Forty- eight SD rats were randomly divided into 6 groups：normal control group, model group and budesonide treatment group, catgut implantation at acupoints for 1,2 and 3 weeks groups. In addition to the normal control group, the rest groups adopted ovalbumin sensitization method to make allergic rhinitis SD rats model. After the success of the building, three groups of catgut implantation were treated at acupoint Yingxiang （ LI20 ） , Quchi （ LI11 ） , Zhusanli （ ST36 ） into a 2 - 0 gut and observe the effect after 1,2 and 3 weeks The budesonide group were given budesonide nasal drop 5 μg/ intranasal （kg · d）, once per day. The model group maintained intranasal ovalbumin and normal group was given intranasal saline solution. Observe the following indexes ： （1）The related behavior score changes ; （2）HE staining ： nasal mucosa pathological morphological changes;（3）By immunohistochemical and pathological methods to test the changes of sub- stance P（SP） and NF- κBp65 protein expression ;（4）By immunofluorescence double staining methods to test the changes of TLR2 and TLR4 protein expression differences in mast cell membrane ;（5）By toluidine blue staining method to test mast cell degranulation rate differences. Results ： Along with the extension of catgut implantation at acupoints, the treatment for 3 weeks can obviously improve the symptoms of AR better than treatment for 1 or 2 weeks and budesonide treatment group,with statistically signifieant（P 〈0. 01 ）. HE staining showed catgut implantation at acupoints and budesonide group can change nasal mucosa cilia state such as lodging, falling off and eosinophil

关 键 词：穴位埋线 变应性鼻炎 神经免疫联动机制 肥大细胞 信号通路 

分 类 号：R245.91[医药卫生—针灸推拿学]