2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/650861摘要：乾燥症是最常見的自體免疫病之一，但是由於臨床上屬於慢性漸進性病程，常常無法在初期即有顯著的症狀，同時又受限於檢驗的敏感性及特異性不足，往往在確定診斷時病情已經達到相當程度的代償不能。乾燥症雖然有一部分患者屬於輕症，但是大多數患者卻會隨著時間健康情形慢慢惡化。因此乾燥症並不是只是單純的口乾、眼乾，伴隨而來的是慢性牙齦炎、蛀牙、口腔黏膜潰瘍、口腔念珠菌感染、胃發炎、食道逆流等逐漸導致消化吸收不良，同時因乾燥嚴重影響睡眠進而衍生出憂鬱及生活品質的惡化。相對的早期診斷治療可以有效改善病程，並降低因長期疾病所衍生的間接醫療成本。同時除了原發性乾燥症外，沒有積極有效治療的其他結締組織病或自體免疫病，常常會隨著時間併發次發性乾燥症，而這些風濕病幾乎涵蓋了目前所有的自體免疫病，其中自體免疫性甲狀腺炎是造成次發性乾燥症最常見的原因。因此乾燥症是長期以來造成健康衝擊又被低估最重要的慢性病之一。受限於臨床檢驗的不足使得乾燥症的早期診斷不易，近年來隨著免疫學的進展發現微小RNA 藉由調控各種訊息RNA 的表現，與細胞的分化、細胞週期、細胞凋亡及免疫功能等都息息相關，也因此微小RNA 的表現可能是形成腫瘤、神經精神病變、病毒感染及各種自體免疫病重要的關鍵過程。在自體免疫病方面初步的研究顯示微小RNA-146a 與類風濕性關節有關、微小RNA-155 與紅斑性狼瘡有關等等。雖然愈來愈多的證據顯示為小RNA 對自體免疫病的重要性，但是對於乾燥症的研究去相當有限，由於微小RNA 也會出現於各種體液，同時研究也顯示唾液中也存在各種微小RNA。因此我們希望藉由對微小RNA 在唾液、血清及免疫細胞的表現及功能來探討原發性及次發性乾燥症的病理機轉、自體免疫抗體在其中的角色，並有助於發展對乾燥症早期診斷的生物標幟及治療的標的。以下是我們預計完成的工作：(A)比較乾燥症、自體免疫性甲狀腺炎及健常人血清、唾液及調控T 淋巴球培養上清液細胞激素的表現(B) 比較不同疾病活性的乾燥症外來體微粒微小RNA 的表現(C) 比較乾燥症、自體免疫性甲狀腺炎及健常人外來體微粒微小RNA 的表現(D)比較乾燥症、自體免疫性甲狀腺炎及健常人血清及單核球細胞微小RNA-155、微小RNA-146a、微小RNA-17-5-p、微小RNA181a 極微小RNA150 的表現(E) 比較乾燥症、自體免疫性甲狀腺炎及健常人藉由調控微小RNA155 再調控T 細胞及培養上清液來探討為小RNA155 的功能(F) 比較自體抗體anti-SSA/SSB 及甲狀腺抗體anti-TPO/TA 對於調控T 細胞及培養上清液微小RNA-155 的影響(G)比較乾燥症、自體免疫性甲狀腺炎及健常人藉由調控微小RNA-146a 再調控T 細胞及培養上清液來探討為小RNA146a 的功能(H)比較自體抗體anti-SSA/SSB 及甲狀腺抗體anti-TPO/TA 對於調控T 細胞及培養上清液微小RNA-146a 的影響我們希望藉由這些實驗來確認乾燥症患者為小RNA 的異常，並釐清其功能及免疫相關的致病機轉，以其有助於將來臨床診斷及治療上的應用。<br> Abstract: Sjogren’s syndrome, characterized by systemic autoimmunity and inflammation anddysfunction in the exocrine organs primarily the salivary and lachrymal glands, is one of themost common systemic autoimmune diseases. Because of subtle clinical manifestation andcourse of slow progression, Sjogren’s syndrome is under-diagnosed for a long time. Agrowing evidence suggest that Sjogren’s syndrome is associated with much long-termcomplication and sometimes life-threatening events including gingivitis, dental caries, oralcandidiasis, atrophy gastritis, poor appetite, malnutrition, sleep disturbance and depression etc.In addition, secondary Sjogren’s syndrome is more common than we known, especially withautoimmune thyroiditis. The exact cause of exocrine dysfunction in Sjogren’s syndrome is notknown but it is likely that both immunologically mediated and non-immune mechanismscontribute significantly, and the related reports are limited.MicroRNAs, 20–22 nucleotide long non-coding RNA molecules, are known to regulatepost-transcriptional regulation of certain subsets of messenger RNAs thus targeting them fordegradation or translational repression. The importance of miRNA regulation to cellularfunctions is becoming increasingly clear including cellular processes such as apoptosis,differentiation, cell cycle, and immune functions. Therefore, microRNAs are associated withautoimmune diseases, and could potentially serve as diagnostic biomarkers and therapeutictargets. For examples, microRNA-146a Contributing to Abnormal Activation of the Type IInterferon Pathway in Human Lupus, microRNA-155 over-expression in lupus regulatory Tcells, and microRNA-155 required for normal immune function, germinal center response,and generation of Ig class-switched plasma cells Inflammatory implicated the critical role ofmicroRNAs in the pathogenesis of autoimmune diseases. But to date, only a relatively smallnumber of specific miRNAs have been revealed as important regulators of the immunesystem.Although the growing body of evidence suggests the critical role of microRNAs inimmune regulation, there are only limited reports about the relevance with autoimmunedisease. In this study, we want to conduct the following experiments to elucidate themicroRNAs defect in the overlooked Sjogren’s syndrome and the associated pathogenesis.(A)Compare the expression of cytokine profiles in plasma, saliva and supernatant of Treg cellculture between Sjogren’s syndrome, autoimmune thyroiditis and healthy control.(B) Compare the microRNA biomarkers from exosomes of saliva in Sjogren’s syndrome withdifferent disease activity and stages by serology and image study(C) Compare the microRNA biomarkers from exosomes of saliva between Sjogren’s syndrome,autoimmune thyroiditis and healthy control(D)Compare the expression of miRNA-155, miRNA-146a, miRNA-17-5-p, MiRNA 181a,miRNA-150 in plasma and mononuclear cells between Sjogren’s syndrome, autoimmunethyroiditis and healthy control.(E) Compare the biologic function of miRNA-155 by modulating the miRNA-155 expressionof Treg cells and culture supernatant between Sjogren’s syndrome, autoimmune thyroiditisand healthy control(F) Compare the biologic effect of anti-SSA/SSB and anti-TPO/TA on miRNA-155 expressionof Treg cells and culture supernatant from healthy control(G)Compare the biologic function of miRNA-146a by modulating the miRNA-146aexpression of Treg cells and culture supernatant between Sjogren’s syndrome, autoimmunethyroiditis and healthy control(H)Compare the biologic effect of anti-SSA/SSB and anti-TPO/TA on miRNA-146aexpression of Treg cells and culture supernatant from healthy controlWe hope that Sjogren’s syndrome relevant defect of microRNAs could be identified andthe functional and immunologic effects might be further elucidated for the pathogenesis afterthese investigations, which would explore the field of diagnosis and therapeutic use.