Alvarez-Berdugo D, Rofes L, Farré R, Casamitjana J F, Enrique A, Chamizo J, Padrón A, Navarro X, Clavé P
Gastrointestinal Motility Laboratory, Hospital de Mataró, Consorci Sanitari del Maresme, Mataró, Spain.
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Instituto de Salud Carlos III, Barcelona, Spain.
Neurogastroenterol Motil. 2016 Jan;28(1):91-100. doi: 10.1111/nmo.12701. Epub 2015 Nov 4.
Previous studies have found that TRPV1 and TRPA1 receptor agonists improve swallow response in patients with oropharyngeal dysphagia (OD), but little is known about the expression of these receptors in the human oropharynx. The aim of this study was to assess the expression and localization of TRPV1 and TRPA1 in human samples from the oropharynx of healthy patients, to provide the basis for new pharmacological treatments for OD.
Samples from oropharyngeal regions innervated by cranial nerves V, IX, and X (tongue, pharynx, and epiglottis) were obtained during ENT surgery and processed either for mRNA (21 patients) or for immunohistochemical assays (seven patients). The expression analysis was performed with RT-qPCR using ACTBh as reference gene. Hemotoxylin and eosin staining was used to study the histology; the immunohistochemical assay used (i) neuron-specific enolase to detect nerve fibers or (ii) fluorescent probes to locate TRPV1 and TRPA1.
TRPV1 was expressed in the three studied regions, with higher levels in CN V region (tongue) than in CN X region (epiglottis; p < 0.05), and was localized at epithelial cells and nociceptive fibers in all studied regions. TRPA1 was also expressed in all studied regions, but was always localized below the basal lamina. No immunoreactivity for TRPA1 was found on epithelial cells.
CONCLUSIONS & INFERENCES: TRPV1 and TRPA1 are widely expressed in the human oropharynx with two distinct patterns. Our study further confirms that TRPV1/A1 receptors are promising therapeutic targets to develop active treatments for OD patients.
先前的研究发现,瞬时受体电位香草酸亚型1(TRPV1)和瞬时受体电位锚蛋白1(TRPA1)受体激动剂可改善口咽吞咽困难(OD)患者的吞咽反应,但对于这些受体在人类口咽部的表达情况知之甚少。本研究的目的是评估TRPV1和TRPA1在健康患者口咽部人类样本中的表达及定位,为OD的新药理学治疗提供依据。
在耳鼻喉科手术期间获取由颅神经V、IX和X支配的口咽区域(舌、咽和会厌)的样本,并对其进行mRNA检测(21例患者)或免疫组织化学分析(7例患者)。使用β-肌动蛋白(ACTBh)作为参考基因,通过逆转录定量聚合酶链反应(RT-qPCR)进行表达分析。苏木精和伊红染色用于研究组织学;免疫组织化学分析使用(i)神经元特异性烯醇化酶检测神经纤维,或(ii)荧光探针定位TRPV1和TRPA1。
TRPV1在所有三个研究区域均有表达,在颅神经V区域(舌)中的表达水平高于颅神经X区域(会厌;p<0.05),并且在所有研究区域均定位于上皮细胞和伤害性纤维。TRPA1在所有研究区域也有表达,但总是定位于基膜下方。在上皮细胞上未发现TRPA1的免疫反应性。
TRPV1和TRPA1在人类口咽部广泛表达,呈现两种不同的模式。我们的研究进一步证实,TRPV1/A1受体是为OD患者开发有效治疗方法的有前景的治疗靶点。