2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/646082摘要：鼻黏膜受神經系統及免疫系統的調控，執行保護下呼吸道的任務。免疫系統的反應緩慢但有特異性，例如鼻黏膜表皮細胞具有各種受體(如Toll-like receptors )偵測外來致病原，繼而分泌抗菌物質(如β-defensin, SPLUNC1 等)；而神經系統的反應迅速，例如打噴嚏、分泌鼻水及擴張血管等。自1950 年代，就發現這兩個系統存在雙向的互動(crosstalk)來對抗外來物的入侵，這種互動主要依賴neuropeptides, neurotrophins 及cytokines,chemokines 等小分子作用，例如神經細胞可分泌cytokines, chemokines 來活化免疫系統，而神經細胞也具有受體可接受其作用(IL-1, IL-2, IL-6 等)；同樣的，免疫細胞也可分泌神經生長激素NGF 影響neural plasticity；免疫細胞同樣具有神經激素的受體(ACTH, PRL,TRH receptors)接受其作用，因而形成完整的互動網絡；這種雙向的溝通形成互補的作用。而鼻黏膜上的表皮細胞是呼吸系統的第一道防線，執行免疫功能。表皮細胞可分泌NGF, BDNF 及IL-6, IL-8, MCP-1 等分子，因此表皮細胞上的神經免疫互動必然存在。但最近發現在老鼠鼻黏膜表皮上有特異的solitary chemosensory cells (SCCs)被緻密的三叉神經末稍包圍，並利用味覺受體(如T2Rs)感知革蘭式陰性菌的bacterial quorum-sensingmolecules，以活化相鄰的神經末稍，改變動物的呼吸型態，1 顯見除了埋藏在表皮細胞間的游離神經末稍可感知外來有害刺激外(如capsaicin、acetic acid)，SCCs 與神經末稍的聯結，也執行固有免疫的保護機制。而在人體呼吸道的纖毛細胞也發現T2R4, T2R43,T2R38, T2R46 等味覺受體，與老鼠的SCCs 雷同，可接受苦味分子的刺激，使纖毛揮動加速，促進排除功能。2 因此，在表皮細胞上呈現的神經免疫互動，更形重要。本計畫將使用下鼻甲黏膜(控制組)，慢性鼻竇炎無鼻息肉之黏膜及鼻息肉以進行下列研究。第一年要釐清的目標，是以免疫染色探究人體的鼻黏膜表皮層有否如老鼠特異分化的chemosensory cells。第二年的目標，以RT-PCR、免疫染色、Flow cytometry、RT-Q-PCR，探查表皮細胞是否具有neuropeptides, neurotrophins,的受體存在；第三年研究的目標，以表皮細胞培養的模式，加入neuropeptides, neurotrophins 刺激表皮細胞，並測量cytokines, chemokines 分泌的改變，或反之亦然；以探討兩系統互動的分子機制。並分別定量三種不同鼻黏膜的表現，以瞭解不同發炎狀態表皮細胞的角色。經由本計畫的研究，希望回答以下問題：表皮細胞是否具有神經傳導物質受體以接受刺激？當表皮脫落壞死，暴露的神經末稍則更容易被刺激，因此在表皮復原過程當中，神經傳導物質是否會促進表皮細胞增生及分化？表皮細胞在發炎狀態是否會分泌神經生長激素導引神經走向？藉以上的研究辨識神經性發炎的生物標記，幫助臨床診斷；並了解鼻黏膜神經性發炎的機轉，進一步尋求可能的治療方法。<br> Abstract: Nasal mucosa is the contact zone that could eliminate small particles and water-solublevolatile compounds from inhaled air so that only clean, humidified air reaches the lowerairway and thus protects the lung. The processes that carry out these functions are under strictregulation by both immunological and neurological systems. The nasal epithelial cellsexpressed receptors (such as Toll-like receptors) which detect potential pathogens. Via signaltransduction, the cells may secret antimicrobial agents such as β−defensin, SPLUNC1. On theother hand, multiple subsets of nociceptive, parasympathetic, and sympathetic nervesinnervate human nasal mucosa. These play carefully coordinated roles in regulating glandular,vascular, and other processes. The crosstalk between immunological and neurological systemshas been discovered since 1950s. Neuronal cells contain receptors that respond to smallmolecules from immunocytes such as cytokines and chemokines (IL-1, IL-2, IL-6 etc.). Viceversa, some immunocytes may secrete nerve growth factors which control neural plasticityand contain neurotransmitter receptors (ACTH, PRL, TRH receptors). Therefore, the crosstalkbetween both systems is mutually complementary.The nasal epithelial cells are the first contact of human airway to external environment.They have been considered as important sensors and effectors for immune systems. Althoughsome evidences showed that nasal epithelial cells may produce NGF and BDNF and theycontain biding sites for SP, GRP, VIP and M1, 3; whether the neuroimmune crosstalk actuallyhappened in nasal epithelial cells is not clear yet. Other than free intra-epithelial nerveendings can detect certain irritants, recent findings from mouse revealed that the epitheliumalso houses a population of trigeminally innervated solitary chemosensory cells (SCCs) thatexpress T2R bitter taste receptors along with their downstream signaling components. TheseSCCs have been postulated to enhance the chemoresponsive capabilities to some irritantsincluding acyl–homoserine lactone bacterial quorum-sensing molecules. Furthermore, motilecilia of human airway epithelia are found to be chemosensory using expressed sensory bittertaste receptors. Therefore, it is now obvious that nasal epithelial cells play much importantrole in both immunological and neurological systems than previously known.At present study, we use nasal mucosa obtained from patients who received operationdiagnosed as chronic hypertrophic rhinitis, chronic paranasal sinusitis without polyps andnasal polyps. The goal of first year is to investigate if solitary chemosensory cells (SCCs) thatexpress T2R bitter taste receptors are also present in human nasal mucosa by usingimmunohistochemical staining. For the second year, the goal is to investigate if neuropeptides,neurotrophins, cytokines, chemokines and their receptors are present in nasal epithelial cellsby using RT-PCR, immunohistochemical staining, flow cytometry and RT-Q-PCR. For thethird year, cultured nasal epithelial cell under air-liquid interface would be used to investigatethe effect of neuropeptides, neurotrophinson the secretion of cytokines, chemokines in nasalepithelial cells.From our study, we might be able to answer the questions: Whether nasal epithelial cellshave receptors respond to neuropeptides or neurotrophins? What is the role of exposed nerveendings to the healing of nasal epithelial cells? Whether neurotransmitter is able to ameliorateproliferation and differentiation of epithelial cells? If epithelial cells would secrete nervegrowth factors to guide the growth of injured nerve endings? This study will help usidentifying the molecular markers and improving our therapeutic methods in treatingneurogenic inflammation.鼻息肉表皮細胞神經生長因子 betaIL-6IL-8MCP-1SPLUNC1神經免疫互動nasal polypepithelial cellsnerve growth factor betaIL-6IL-8MCP-1SPLUNC1neuroimmune crosstalk