Mendonça Jeferson Cedaro de, Dolci José Eduardo Lutaif
Department of Otorhinolaryngology, Santa Casa de Sao Paulo, Faculdade de Ciências Médicas.
Braz J Otorhinolaryngol. 2005 Mar-Apr;71(2):123-31. doi: 10.1016/s1808-8694(15)31299-4. Epub 2005 Aug 2.
Neuropeptides are important neurotransmitters in nasal physiology and the increasing knowledge of their role in nasal diseases brings new therapeutic perspectives. The investigation of human nasal mucosa neuropeptides is based mostly on immunocytochemistry, a complex approach whose resulting factors may be variable. Aiming to make this kind of research available, an immunofluorescence approach for vasoactive intestinal peptide (VIP) in human nasal mucosa is proposed and evaluated.
Transversal cohort.
Human inferior turbinate samples were obtained at time of nasal surgery from eight patients. The samples were fixed in Zamboni solution (4% phosphate-buffered paraformaldehyde and 0.4% picric acid), snap-frozen and stored at -70 degrees C. 14 microm sections were then obtained. Immunofluorescence staining for VIP (Peninsula Laboratories) was performed and its images documented by conventional photography. The method's specificity, sensitivity and reproducibility of execution were evaluated. Additionally, the reproducibility of interpretation of results was evaluated through the comparison of staining scores (0 to 4) attributed to the images by six observers.
The results showed the approach to be very specific and sensible, besides being reproducible in its execution. The interpretation of results may depend on the observer's accuracy in judging immunofluorescence images, but it showed uniformity.
The proposed method was highly useful for research purposes in neuropeptides in human nasal mucosa.
神经肽是鼻腔生理学中的重要神经递质，对其在鼻腔疾病中作用的认识不断增加带来了新的治疗前景。对人鼻黏膜神经肽的研究主要基于免疫细胞化学，这是一种复杂的方法，其结果因素可能存在差异。为了使这类研究可行，本文提出并评估了一种用于检测人鼻黏膜中血管活性肠肽（VIP）的免疫荧光方法。
横向队列研究。
在鼻手术时从8名患者获取下鼻甲样本。样本用赞博尼溶液（4%磷酸盐缓冲多聚甲醛和0.4%苦味酸）固定，速冻后储存在-70℃。然后获得14微米厚的切片。进行VIP（半岛实验室）的免疫荧光染色，并用传统摄影记录其图像。评估该方法的特异性、敏感性和执行的可重复性。此外，通过比较6名观察者对图像给出的染色评分（0至4）来评估结果解释的可重复性。
结果表明该方法非常特异且灵敏，执行过程具有可重复性。结果的解释可能取决于观察者判断免疫荧光图像的准确性，但显示出一致性。
所提出的方法对人鼻黏膜神经肽的研究目的非常有用。
