P物质在组胺鼻激发诱导眼部变应性炎症中的作用初探  

A preliminary study on the role of substance P in histamine-nasal-spray-induced allergic conjunctivitis in guinea pigs

作　　者：李彤[1] 赵长青[1] 

机构地区：[1]山西医科大学第二医院耳鼻咽喉头颈外科,太原030001

出　　处：《中华耳鼻咽喉头颈外科杂志》2015年第10期836-841,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金（81271059）;山西省基础研究项目（2012011039-1）;山西省留学回国人员科研资助项目（2014-重点-3）

摘　　要：目的借助豚鼠组胺鼻激发诱导结膜变应性炎性反应过程，探讨非肾上腺素能非胆碱能神经（non-adrenergic noncholinergic nerve，NANC）及其介质P物质（substance P，SP）在此炎性反应过程中可能存在的作用及对肥大细胞（mastcell，MC）和嗜酸粒细胞（eosinophil，Eos）的影响。方法雄性豚鼠40只利用随机数字表随机分为A—E5组，每组8只，A组为空白对照组，B组为单纯组胺激发组，C、D、E各组序贯阻断胆碱能、肾上腺素能神经及SP受体后，一侧鼻腔滴注组胺，30min后观察同侧结膜血管通透性和组织学、MC、Eos形态与功能以及SP分泌的变化。结果运用SPSS17．0统计软件进行分析。结果组胺鼻激发能够引起同侧结膜血管通透性增大及sP释放增多；胆碱能神经阻滞剂阿托品、sP受体阻滞剂能够抑制上述变化。A、B、C、D、E组同侧结膜组织伊文蓝含量分别为（13．78±2．48）、（29．62±3．31）、（19．03±1．47）、（18．42±2．52）、（14．83±2．14）μg/ml，A组与B组，B组与c组，c组与E组相比，差异有统计学意义（t值分别为-10．66、7．97、4．51，P值均〈0．05）。5组同侧结膜组织中SPmRNA的相对表达量分别为（1．00±0．04）、（1．61±0．09）、（1．26±0．03）、（1．27±0．06）、（1．08±0．05），A组与B组，B组与C组，C组与E组相比，差异有统计学意义（t值分别为-22．04、12．93、11．85，P值均〈0．05）。5组同侧结膜组织中组织类胰蛋白酶mRNA的相对表达量分别为（1．00±0．01）、（1．01±0．05）、（1．02±0．17）、（1．00±0．14）、（1．01±0．20），嗜酸粒细胞主要碱性蛋白1mRNA的相对表达量分别为（1．00±0．03）、（1．02±0．15）、（0．94±0．08）、（1．01±0．07）、（0．98±0．13），各组比较差异均无统计学意义（F值分别为1．93、0．57，P值均〉0．05）。结论豚鼠组胺滴鼻可以诱发Objective To investigate the effect of the non adrenergic non cholinergic nerve （NANC） and substance P （SP） in allergic rhinoconjunctivitis by observing histamine nasal provocation induced conjunctivitis in guinea pigs. Methods Forty male guinea pigs were randomly divided into five groups with each group consisting of eight guinea pigs. All anesthetized guinea pigs were exposed either to histamine （0.2%, 5 μl） （group B - E） or saline （5 μl, group A） via unilateral nostril. No pretreatment was done in group A and B while pretreatment was done in groups C - E through injection into the unilateral common carotid artery with eholinergic nerve inhibitor （ atropine, 1 mg/kg, group C ）, cholinergic nerve inhibitor plus adrenergic nerve inhibitors （ atropine, 1 mg/kg, phentolamine, 1 mg/kg plus Esmolol, 1 mg/kg, group D） and cholinergic nerve inhibitor, adrenergic nerve inhibitors plus SP receptor antagonist （the same treatment with group D plus D-SP 10-6mol/L, 1 μl/g, group E）, respectively. To assess the ipsilateral conjunctival inflammatory reaction, conjunctiva leakage with Evans blue dye assessments and HE staining of conjunctival tissues were performed. The SP expression in ipsilateral conjunctival tissue in different groups of guinea pigs were assessed by immunofluorescence and RT-PCR. The activity of eosinophils was assessed by eosinophil major basic protein 1 （MBP1） with RT-PCR, meanwhile, the activity of mast cells was assessed by tryptase with RT-PCR. SPSS 17.0 software was used to analyze the data. Results At 30 min after nasal application of histamine, ipsilateral conjunctivitis was successfully induced as shown by the change of conjunctiva leakage and histology. The content of Evans blue in ipsilateral conjunctival tissue of group A - E was （13.78±2.48）, （29.62±3.31）, （19.03 ±1.47）, （18.42 ±2.52）, （14.83 ±2. 14） μg/ml, respectively. There was statistically significant difference between group A and B （ t = - 10.66, P 〈 0.05 ） , group B a

关 键 词：鼻激发试验 结膜炎 变应性 P物质 速激肽类 

分 类 号：R765[医药卫生—耳鼻咽喉科]