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

The effects of static magnetic fieldson lipopolysaccharide and lipoteichoic acid induced excessive immune reaction–in vitro investigation on fibroblast, and osteoblast-like cells

陳俊仰(Chun-Yang Chen)

指導教授 ： 林哲堂教授 ； 李勝揚 教授 ； 黃豪銘 副教授

臺北醫學大學/口腔學院/牙醫學系碩博士班/碩士(2006年)

摘要

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。

關鍵字

靜磁場 ； 纖維母細胞株 ； 似骨母細胞株 ； 脂多醣 ； 脂壁酸

並列摘要

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.

並列關鍵字

static magnetic fields ； fibroblast ； osteoblast ； lipopolysaccharide(LPS) ； lipoteichoic acid(LTA)

參考文獻

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