2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38610In Taiwan, there are two million people who have the betel quid chewing habit, which has been suspected to elevate the incidence of oral submucous fibrosis (OSF). OSF, which is regarded as a precancerous condition, is characterized by juxta-epithelial inflammatory reaction followed by fibro-elastic change in the lamina properia and epithelial atrophy. The connective tissue changes are characterized by deposition of dense collagen fibers in the oral mucosa. Most of these studies were focused on the changes in the collagen accumulation in connective tissue layer. To date there has been little research into the roles of type I protease activated receptor (PAR-1) in normal buccal mucosa as well as OSF. In this study, immunolocalization of PAR-1 will be detected in human human normal buccal mucosa and submucous fibrosis specimens. In addition, RT-PCR, Western blot and ELISA assays will be used to investigate the roles of PAR-1 on the fibroblasts cultured from human normal buccal mucosa and submucous fibrosis. Furthermore, the possible regulatory mechanisms will also be evaluated by addition with arecoline as well as pharmacologic agents to realize the possible signal transduction pathway of PAR-1. Furthermore, the blood will be corrected from OSF patients and age-matched normal persons. Real time polymerase chain reaction will perform for evaluation of PAR-1 polymorphism.嚼食檳榔是導致口腔黏膜下纖維化症(Oral submucous fibrosis，簡稱OSF)的主要原 因，臨床上最常發生於頰黏膜，其症狀為黏膜僵硬，主要是由於上皮下方結締組織中含 有過量的膠原蛋白沉積所造成，而目前研究絕大多數都朝此方面做探討，第一型蛋白酵 素活化接受器(type I protease activated receptor; PAR-1)在fibrosis 中扮演著非常重要的角 色，鮮少有研究探討PAR-1 與OSF 間的關聯性，吾人認為PAR-1 在OSF 病程中所扮演 的角色值得進一步探討，本研究擬分三年來探討PAR-1 在正常口腔黏膜與OSF 組織間 的差異性。第一年：收集人類正常頰黏膜及口腔黏膜下纖維化組織病理切片，以免疫組 織化學染色法，了解PAR-1 的表達情形。以組織培養法，培養人類正常頰黏膜及口腔黏 膜下纖維化纖維母細胞，抽取RNA、protein 及condition medium，儲存於-70°C 冰箱， 待收集至足夠數目後以利第二年比較PAR-1 分泌量。第二年：以組織培養法，培養人類 正常頰黏膜及口腔黏膜下纖維化纖維母細胞，抽取RNA、protein 分析比較不同層次 PAR-1 含量，以RT-PCR、western blot、ELISA 等方法比較正常頰黏膜及口腔黏膜下纖 維化纖維母細胞PAR-1 分泌量。從流行病學調查研究顯示，嚼食檳榔是導致OSF 的主 要原因，arecoline 是檳榔植物鹼中含量最多的成分，已被證實與OSF 形成有關有密切的 關連，以組織培養法，培養人類正常頰黏膜造纖維母細胞，以RT-PCR、western blot、 ELISA 等方法比較在arecoline 作用下正常頰黏膜造纖維母細胞是否會增加PAR-1 分泌 量。第三年：另外則針對OSF 及正常人之間，探討其間PAR-1 的差異性是否和PAR-1 DNA polymorphism 有關？抽取OSF 病人及正常人血液中genomic DNA，以real time-polymerase chain reaction 之方法來探討PAR-1 polymorphism 的關聯性，並分析是否 和性別、年齡、嚼食檳榔時間長短等因子具有相關性。Project