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研究生:陳俊仰
研究生(外文):Chun-Yang Chen
論文名稱:靜磁場對脂多醣和脂壁酸引發的細胞過度免疫反應之影響-對纖維母細胞、似骨母細胞之體外實驗
論文名稱(外文):The effects of static magnetic fieldson lipopolysaccharide and lipoteichoic acid induced excessive immune reaction–in vitro investigation on fibroblast, and osteoblast-like cells
指導教授:林哲堂林哲堂引用關係李勝揚李勝揚引用關係黃豪銘
指導教授(外文):Che-Tong LinSheng-Yang LeeHaw-Ming Huang
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:牙醫學系
學門:醫藥衛生學門
學類:牙醫學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:110
中文關鍵詞:靜磁場纖維母細胞株似骨母細胞株脂多醣脂壁酸
外文關鍵詞:static magnetic fieldsfibroblastosteoblastlipopolysaccharide(LPS)lipoteichoic acid(LTA)
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近十年來細菌性敗血症和敗血性休克的病例逐年增加,而且死亡率很高。科學界已發現,革蘭氏陰性菌和革蘭氏陽性菌細胞壁成分的LPS和LTA均是引發宿主過度免疫反應的重要毒性物質,而且來自不同菌種的LPS和LTA對不同細胞也會產生不同程度的發炎反應。由於關於此方面的藥物開發,始終沒有醫學臨床上突破的發現,因此尋求物理性非侵入性的治療方法為臨床醫學上的另一思考方向。本研究探討在靜磁場的作用下,是否可廣效的降低來自不同菌種的LPS與LTA對軟硬組織細胞的毒殺作用。本實驗將LPS和LTA加入NIH-3T3、MG63細胞中,尋找內毒素毒殺細胞的能力及 50%的細胞致死濃度。接著加入靜磁場作為物理性的刺激,觀察細胞數量和釋放發炎激素如IL-1、IL-6、TNF-的量。本研究結果顯示,來自不同菌種的LPS和LTA對NIH-3T3與MG63細胞的50%致死濃度均不相同,與文獻的結果相符合。若加入更多的內毒素,細胞的死亡率會達高原期而不再變化。在接續的實驗中,我們發現將NIH-3T3和MG63細胞預先暴露於靜磁場(4000G) 12小時後,可以有效的抑制來自不同菌種的LPS和LTA之細胞毒性。而且靜磁場的預暴露對未加入LPS和LTA的NIH-3T3和MG63細胞沒有毒殺作用。此結果讓我們確定利用靜磁場廣效的抑制LPS或LTA對細胞的過度免疫反應是一個可行的研究方向。
For nearly a decade, cases on sepsis and septic shock have gradually increase over the years. It has been observed that both LPS and LTA compositions found in the cell wall of Gram negative bacteria and Gram positive bacteria were the toxic substances responsible for inducing cytotoxicity. Endotoxin (LPS and LTA) from different bacteria were found to induce different degrees of inflamed responses. However, drugs developed based on the above mechanism showed no significant clinical breakthrough. The continuous search for physiologic non-invasive treatments have been considered as alternative methods. This study aims to investigate the probability of decreasing LPS and LTA cell toxicity on soft and hard tissue when exposed to static magnetic field. To find the toxicity concentrations with 50% cell mortality rate, both LPS and LTA were added into tested cells (NIH-3T3 and MG63 cells) with various concentrations and cell viability were tested by MTT method. To test the inhibitive effects of static magnetic field on the cell count and the amount of inflammatory substances release such as IL-1, IL-6, TNF- etc, NdFeB magnets were used as a physiologic stimulator. The result of this study indicates that 50% cell mortality rate dosage of NIH-3T3 and MG63 cells induced by LPS and LTA from various bacteria were different. This result is consistence to previous investigations. Cell mortality rate reaches a plateau with no further changes when more endotoxins were added. In the latter part of the study, it was observed that when NIH-3T3 and MG63 cells were exposed to a static magnetic field (4000G) for 12 hours, static magnetic field can effectively inhibit the toxic reaction of LPS and LTA with a bacteria-independent manner. On the other hand, NIH-3T3 and MG63 cells exposed to static magnetic field for 12 hours but without the addition of LPS and LTA exhibited no toxic effect. Based on the primary results, we are certain that further research on static magnetic field in inhibiting LPS or LTA induced cytotoxicity is feasible.
目 錄
頁數
中文摘要……………………………………………I
英文摘要…………………………………………………II
目錄……………………………………………………VI

第一章、緒論
第一節 研究動機與重要性……………………1
第二節 研究目的………………………………4
第三節 研究假設…………………………………4
第四節 名詞界定…………………………………5

第二章、文獻回顧與查證
第一節 LPS和LTA與細胞膜結合機制之探討…12
第二節 LPS和引發發炎反應之機制……………13
第三節 內毒素耐受性…………………………16
第四節 來自不同細菌來源的LPS和LTA所引發的
發炎反應……………………………17

第三章、材料與方法
第一節 靜磁鐵之製備及靜磁場之檢測…………19
第二節 實驗之細胞培養方法……………………19
第三節 LPS和LTA對細胞影響之影響………23
1.LPS濃度對細胞生長的影響…………23
2.LTA濃度對細胞生長的影響…………24
第四節 細胞活性的測量
1.以MTT分析法分析靜磁場對LPS毒殺細胞
作用的影響……………………………24
2.以MTT分析法分析靜磁場對LTA毒殺細胞作
用的影響………………………………25
第五節 基因晶片的偵測………………………27

第四章、研究結果……………………………………28

第五章、研究討論...………………………………47

第六章、未來研究目標………………………………53

第六章、參考文獻……………………………………55
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