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    National Tsing Hua University Institutional Repository > 生命科學院  > 分子醫學研究所 > 博碩士論文 >  腺核?酸對細胞移動、緊密連結與細胞毒性之影響

    Please use this identifier to cite or link to this item: http://nthur.lib.nthu.edu.tw/dspace/handle/987654321/85639

    Title: 腺核?酸對細胞移動、緊密連結與細胞毒性之影響
    Authors: 陳郁欣
    Chen, Yu-Hsin
    Description: GH02101080520
    Date: 2014
    Keywords: 腺?酸激?;腺核?酸;綠膿桿菌;緊密連結;細胞毒性;細胞移動
    Adenylate Kinase;Adenine Nucleotides;Pseudomonas aeruginosa;Tight Junction;Cytotoxicity;Cell Migration
    Abstract: 腺核?酸的水解與合成,在細胞代謝反應中具有重要地位。腺?酸激? (Adenylate Kinase)控制不同腺核?酸在細胞內濃度的平衡,普遍存在於真核和原核細胞中。但根據之前的文獻指出,綠膿桿菌 (Pseudomonas aeruginosa) 和鼠疫耶爾森氏菌(Yersinia pestis) 會利用分泌的腺?酸激?當作毒力因子,藉著產生高量的腺核?酸雙磷酸活化巨噬細胞上的嘌呤受器(purinergic receptors),導致巨噬細胞死亡。另外傷口的細胞受到損害時會釋出腺核?酸,當作吸引巨噬細胞、引發免疫反應並調控一連串傷口修復的信號。因此本研究模擬傷口形成後,分別在大腸癌細胞C2BBe1與HCT-8中添加腺核?酸,發現不同的腺核?酸對不同細胞有不同的促進細胞移動的能力。利用綠膿桿菌B136-33上清液與腺核?酸,發現上清液與ATP同時作用造成巨噬細胞RAW 264.7大規模的死亡,而且純化後的腺?酸激?能增加ADP對RAW 264.7的細胞毒殺能力。本研究中發現綠膿桿菌培養後之上清液能夠導致C2BBe1細胞緊密連結蛋白的位移,只加入腺核?酸則能夠讓細胞間緊密連結蛋白相互分離,但綠膿桿菌培養後之上清液與腺核?酸會讓C2BBe1細胞造成劇烈破壞,緊密連結蛋白位移程度卻不像其他組別嚴重。綜合上述實驗結果,證實綠膿桿菌會利用存在細胞外的腺核?酸對上皮細胞與巨噬細胞進行破壞,以達到感染宿主之目的。
    Cells catabolize the nutrient molecules and store the free energy in chemical forms such as ATP. ATP releases its chemical energy through catalysis into 2ADP + Pi. Adenylate kinase, an enzyme catalyzing interconversion of AMP, ADP and ATP, is responsible for maintaining the adenine nucleotide balance in cells. It is found in both eukaryotic and prokaryotic cells. In has been reported previously that adenylate kinase is secreted by Pseudomonas aeruginosa and causes toxicity to macrophage during infection and is likely an important virulence factor. During infection, damaged cells could release adenine nucleotides that may participate bacterial infections, regulate the wound healing, and trigger immune reaction of macrophage to clear the infection. This study found that different adenine nucleotides can enhance the migration ability of C2BBe1 and HCT-8 epithelial cells. Massive cell death of macrophage RAW 264.7 could be observed after treatment with P.aeruginosa isolate B136-33 supernatant supplemented with ATP. Moreover, purified adenylate kinase increases the ADP-mediated RAW 264.7 cell death. My study also showed that P. aeruginosa culture supernatant and adenine nucleotides can alter the structure of tight junction and lead to serve damage of C2BBe1 cell monolayer. Together, our finding confirms previous notions that the adenine nucleotides play an important role in cell physiology and bacterial infections.
    URI: http://nthur.lib.nthu.edu.tw/dspace/handle/987654321/85639
    Source: http://thesis.nthu.edu.tw/cgi-bin/gs/hugsweb.cgi?o=dnthucdr&i=sGH02101080520.id
    Appears in Collections:[分子醫學研究所] 博碩士論文

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