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研究生:陳凱莉
研究生(外文):Kai-Li Chen
論文名稱:探討突變型鏈球菌侵入人類牙齦纖維母細胞誘導發炎因子之表現
論文名稱(外文):Investigate the inflammatory gene expression on human gingival fibroblasts through Streptococcus mutans invasion
指導教授:陳政男陳政男引用關係
指導教授(外文):Cheng-Nan Chen
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生化科技學系研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
畢業學年度:100
語文別:中文
論文頁數:78
中文關鍵詞:突變型鏈球菌
外文關鍵詞:Streptococcus mutans
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過去的研究指出,有口腔疾病的病患中,蛀牙大概占54.1%,對冷熱敏感者占25%,有牙結石問題者占23%。這些問題絕大多數跟牙菌斑有關,細菌利用食物殘渣中的澱粉及葡萄糖作為營養的來源,分解雙糖成為單糖 (葡萄糖),以致產生某些酸性物質,這些酸性物質又與牙齒表面接觸,這種酸蝕現象溶解牙齒的表面 (琺瑯質),使牙齒表面受損,形成牙洞。若不予及時治療,當牙齒嚴重受損,甚至侵犯至牙髓時,口腔中的細菌會因此透過牙髓腔中的血管進入人體,進而導致發炎,嚴重的話將導致死亡或成植物人。
文獻指出,突變型鏈球菌會誘導前發炎調節蛋白大量的表現,如腫瘤壞死因子-α(Tumor necrotic factor-α,TNF-α),而TNF-α能誘導內皮細胞的發炎相關蛋白Intracellular Cell Adhesion Molecule-1 (ICAM-1) 、 Vascular Cell Adhesion Molecule-1 (VCAM-1) 、環氧化酶-2 (Cyclooxygenase-2,COX-2),介白素(Interleukin,IL 如:IL-1、IL-8)的表現。因此在蛀牙狀態下,牙齦組織通常會產生發炎現象,但對於發炎反應是否受突變型鏈球菌侵入的高低有影響卻仍不清楚。故本研究想了解當突變型鏈球菌在侵入人類牙齦纖維母細胞(Human gingival fibroblast; HGF)後的發炎反應,探討其發炎因子環氧化酶-2 之表現,並進一步探討其發病原因及機制。本研究之檢體取至於21位蛀牙病患之21種突變型鏈球菌菌株(Streptococcus mutans,S.mutans),並以MOI=20的S.mutans侵入HGF細胞後探討其侵入毒性之高低,結果顯示編號三號及五號病患的S.mutans侵入性較低,而編號八號及十一號侵入性較高,因此本研究將進一步探討其四位病患在侵入毒性有明顯差異之下,其COX-2、IL-1、IL-8、VCAM、ICAM的表現是否也有顯著的差異。

Tooth decay account for 54.1% of patients with oral diseases; those affected by sensitive teeth account for 25% and dental calculus account for 23%. Plaque is the main cause factor. Tooth decay starts with plaque - a thin, sticky substance that forms naturally on all teeth. Plaque contains bacteria which react with sugars and starches present in our daily diet to form acid substances that are harmful to teeth. If plaque isn’t regularly removed, it will attach to teeth. The plaque bacteria produce acid substance, which will enrode tooth's protective enamel surface. Eventually, acid erodes the soft layer of the tooth resulting in holes or cavities.
The formation of dental caries is a dynamic process. Demineralization and remineralization have a crucial impact on the hardness and strength of tooth enamel. Healthy tooth dependents upon the balance between demineralization and remineralization. Acid oral environment which is undersaturated with mineral ions, relative to a tooth’s mineral content will cause demineralization. The enamel crystal, which consists of carbonated apatite, is dissolved by organic acids (lactic and acetic) that are produced by the cellular action of plaque bacteria in the presence of dietary carbohydrates. Remineralization allows the subsequent loss of calcium, phosphate, and fluoride ions to be replaced by fluorapatite crystals.
Dental caries are dangerous and can cause extreme illness when it other vital organs are affected. Tooth infection can affect major organs such as heart and brain. Also, it can affects every other organ when the infection is in the blood. In extreme cases, the untreated infection can eventually lead to death. Literature indicates that dental caries induced endotoxemia elevates level of serum pro-inflammatory cytokines such as interleukin-1 beta 、 tumor necrosis factor-alpha. ICAM-1 (Intracellular Cell Adhesion Molecule-1)、VCAM-1 (Vascular Cell Adhesion Molecule-1)、COX-2, Interleukin EX. IL-1、IL-8 expression. Under the decay condition, the gum tissue usually produces inflammation. However, it remains unknown whether tooth decay bacteria invasion of human gingival fibroblasts (HGF) is related to inflammation inducement.
The purpose of the study is to investigate the level of inflammation after tooth decay bacteria invade HGF causing expression of inflammatory factor cyclooxygenase -2 (COX-2). We use dental caries plaque clinical samples which were taken from 21 patients of the bacteria number with MOI = 20 (OD660 = 1 is equivalent to 1 ×109) to invade HGF cell to explore the level of toxicity. The results showed that the number 3 and 5 patients with dental caries plaque invasivity is low, and the number 8 and 11 patients with dental caries plaque invasivity is hight. Therefore, this study will further explore the invasive toxicity discrepancy between these four patients and COX-2、IL-1、IL-8、VCAM、ICAM performance expression.

目錄
中文摘要 ……………………………………………………II
英文摘要 …………………………………………………IV
致謝 …………………………………………………………V
目錄 ………………………………………………………VII
圖目錄 ………………………………………………………IX
結果目錄 …………………………………………………X
第一章 緒論(Introduction)
1.1研究動機…………………………………………………… 1
1.2研究目的…………………………………………………… 3
第二章 文獻探討
2.1口腔與微生物 ………………………………………… 4
2.2齲齒 ……………………………………………………… 5
2.3 Streptococcus mutans突變型鏈球菌……………………… 7
2.4 lactobacilli乳酸桿菌 …………………………………… 10
2.5慢性發炎 ………………………………………………… 12
2.6發炎因子 ………………………………………………… 13
2 .7 Nuclear Factor-Kappa B ……………………………… 14
2.8 Interleukin-1………………………………………………… 18
2.9Interleukin-8 …………………………………………… 20
2.10 黏附分子(cellular adhesion molecules) ……………… 22
2.11細胞間黏附因子-1(ICAM-1)……………………… 23
2.12血管細胞黏附因子-1(VCAM-1) …………………… 24
2.13 Cyclooxygenase-2 COX-2…………………………… 25
第三章 研究方法及步驟 (Materials and Methods)
3.1實驗材料 …………………………………………………26
3.2實驗方法 …………………………………………………32
第四章 實驗結果(Results) ………………………………44
第五章 討論(Discussiom) ………………………………50
參考文獻 …………………………………………………… 52
附錄…………………………………………………………59

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