| Objective: Allergic rhinoconjunctivitis is a common disorder.Symptoms of conjunctivitis(tearing and itching) occur frequently in patients with allergic rhinitis.High prevalence of allergies rhinoconjunctivitis was observed in many developed countries,while there isn’t relevant statistical data in our country.The pathogenesis underlying these symptoms remains to be elucidated.We hypothesize that these symptoms arise via a combination of mechanisms,including direct contact of natural pollen with the the nasal mucosa and conjunctiva and reflex mechanisms of the nasal-nasal and nasal-ocular reflex.The aim of the present study was to study the epidemiology of allergic conjunctivitis in allergic rhinitis patients from Taiyuan,China,and to analyze the correlation of clinical characteristics between allergic rhinitis and allergic conjunctivitis.On the other hand,to investigate the effect of the nasal-nasal and nasal-ocular reflex in allergic rhinoconjunctivitis by observing histamine nasal provocation induced rhinoconjunctivitis in guinea pigs. Methods: 1 Clinical epidemiological investigation A questionnaire survey of 80 allergic rhinitis patients from the city of Taiyuan was carried out.Their clinical situations and treatment status were investigated in detail,and a database was set up for statistics. 2 nasal-nasal reflex and neurogenic inflammation 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%,5μl)(group 2~5) or saline(5μl,group 1) via unilateral nostril.No pretreatment was done in group 1 and 2 while pretreatment were done in groups 3~5 through injection into the unilateral common carotid artery with cholinergic nerve inhibitor(atropine,1mg/kg,group 3),cholinergic nerve inhibitor plus adrenergic nerve inhibitors(atropine,1mg/kg;phentolamine,1mg/kg plus Esmolol,1mg/kg;group 4) and cholinergic nerve inhibitor,adrenergic nerve inhibitors plus SP receptor antagonist(the same treatment with group 4 plus D-SP 1×10-6mol/L,1ul/g,group 5),respectively.To assess the both sides of nasal mucosal inflammatory reaction,leakage of nasal mucosa with Evans blue dye assessments and HE staining of nasal mucosal tissues were performed.The SP expression in the both sides of nasal mucosal tissues in different groups of guinea pigs were assessed by immunofluorescence,RT-PCR and Elisa.The activity of eosinophils in the both sides of nasal mucosal tissues in different groups of guinea pigs were assessed by HE staining,quantifing the expression of eosinophil cationic protein(ECP) using Elisa and eosinophil major basic protein 1(MBP1) with RT-PCR,meanwhile,the activity of mast cells were assessed by toluidine blue staining quantifing the expression of tumor necrosis factor-(TNF-) using Elisa and Tryptase with RT-PCR.SPSS 17.0 software was used to analyze the data. 3 nose- eye reflex and neurogenic inflammation 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%,5μl)(group 2~5) or saline(5μl,group 1) via unilateral nostril.No pretreatment was done in group 1 and 2 while pretreatment were done in groups 3~5 through injection into the unilateral common carotid artery with cholinergic nerve inhibitor(atropine,1mg/kg,group 3),cholinergic nerve inhibitor plus adrenergic nerve inhibitors(atropine,1mg/kg,phentolamine,1mg/kg plus esmolol,1mg/kg,group 4) and cholinergic nerve inhibitor,adrenergic nerve inhibitors plus SP receptor antagonist(the same treatment with group 4 plus D-SP 1×10-6mol/L,1ul/g,group 5),respectively.To assess the ipsilateral nasal mucosal and conjunctival inflammatory reaction,leakage of conjunctiva and nasal mucosa with Evans blue dye assessments and HE staining of conjunctival and nasal mucosal tissues were performed.The SP expression in ipsilateral conjunctival and nasal mucosal tissues in different groups of guinea pigs were assessed by immunofluorescence,RT-PCR and Elisa.The activity of eosinophils in ipsilateral conjunctival and nasal mucosal tissues in different groups of guinea pigs were assessed by HE staining,quantifing the expression of eosinophil cationic protein(ECP) using Elisa and eosinophil major basic protein 1(MBP1) with RT-PCR,meanwhile,the activity of mast cells were assessed by toluidine blue staining,quantifing the expression of tumor necrosis factor-(TNF-) using Elisa and Tryptase with RT-PCR. SPSS 17.0 software was used to analyze the data. Results 1 Clinical epidemiological investigation In 80 allergic rhinitis patients,66(82.5%) were accompanied with allergic conjunctivitis.The episode ages and their courses of allergic rhinitis and allergic conjunctivitis had no significant differences( P>0.10).In respect of severity of allergic rhinitis in 66 patients,29 were mild,29 moderate and 8 severe.In respect to severity of allergic conjunctivitis,29 were mild,29 moderate and 8 severe. 2 nasal-nasal reflex and neurogenic inflammation At 30 min after one nasal application of histamine,bilateral rhinitis was successfully induced as shown by the change of nasal mucosa leakage and histology.Nasal mucosa leakage and SP expression were significantly higher in group 2 than in group 1(all P <0.05).Pretreatment with cholinergic nerve inhibitor(group 3) reduced the bilateral nasal mucosa leakage and the SP expression than in group 2 while pretreatment with cholinergic nerve inhibitor, adrenergic nerve inhibitors plus SP receptor antagonist(group 5) reduced the bilateral nasal mucosa leakage and the opposite SP expression than in group 3(all p<0.05).The levels of ECP,TNF- and the expression of MBP1 and tryptase of bilateral nasal mucosa in all groups were not significantly different(p>0.05). 1) nose- eye reflex and neurogenic inflammation At 30 min after one nasal application of histamine,ipsilateral conjunctivitis was successfully induced as shown by the change of conjunctiva leakage and histology. Conjunctiva leakage and SP expression were significantly higher in group 2 than in group 1(all P <0.05).Pretreatment with cholinergic nerve inhibitor(group 3) reduced the ipsilateral conjunctiva leakage and the SP expression than in group 2 while pretreatment with cholinergic nerve inhibitor, adrenergic nerve inhibitors plus SP receptor antagonist(group 5) reduced the ipsilateral conjunctiva leakage and the SP expression than in group 3(all p<0.05).The levels of ECP,TNF- and the expression of MBP1 and tryptase of ipsilateral conjunctiva in all groups were not significantly different(p>0.05). Conclusions 1 The morbidity of allergic conjunctivitis in allergic rhinitis patients is high in Taiyuan,China.There are correlations between clinical characteristics of allergic rhinitis and allergic conjunctivitis,and their severity are highly coherent. 2 Histamine provocation in one nasal induced allergic inflammatory response of opposite nasal mucosa and ipsilateral conjunctiva in guinea pigs. 3 Neural factors including parasympathetic nerve,NANC nerves and its medium SP participated this nose-nose and nose-ocular reflex process. |
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