非營養性甜味劑對於滋養層細胞球體植入子宮內膜細胞的影響

Taipei Medical University Institutional Repository > 營養學院 > 保健營養學系暨研究所 > 博碩士論文 > Item 987654321/62302

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題名:	非營養性甜味劑對於滋養層細胞球體植入子宮內膜細胞的影響 Effects of non-nutritive sweeteners on trophoblast spheroids implantation in endometrial cells
作者:	陳憬儀 CHEN, JING-YI
貢獻者:	保健營養學系碩士班謝榮鴻
關鍵詞:	非營養性甜味劑;滋養層細胞;子宮內膜細胞;粒線體;活性氧物種;球體附著試驗 non-nutritive sweeteners;trophoblast;endometrial cells;mitochondria;reactive oxygen species;spheroid-adhesion
日期:	2022-07-05
上傳時間:	2022-12-15 15:31:46 (UTC+8)
摘要:	不孕症為全球關注的議題之一，懷孕期間暴露於非營養性甜味劑使活性氧物種的生成增加及降低粒線體功能，影響胎盤及胎兒的發育，進而阻礙著床而無法受孕。本次研究旨在探討阿斯巴甜、蔗糖素及醋磺內酯鉀對於子宮內膜細胞及與滋養層細胞球體附著之影響。分別以0.5、1 及 2 mM 阿斯巴甜、蔗糖素及醋磺內酯鉀培養人類子宮內膜腺癌細胞 24 小時，以螢光染色分析細胞與粒線體氧化壓力、粒線體膜電位及其數量，以海馬生物能量測定儀分析粒線體呼吸耗氧速率以評估粒線體功能，以免疫呈色法分析抗氧化酵素活性，以西方墨點法分析系線體生合成相關蛋白表現量。另外，將滋養層細胞球體與子宮內膜細胞共培養，評估球體附著率作為體外著床模型。結果顯示，蔗糖素會降低滋養層球體附著率，藉由增加其細胞與粒線體中的氧化壓力，降低粒線體SOD活性，抑制了粒線體呼吸功能，增加粒線體膜電位，並代償性增加粒線體數量。阿斯巴甜及醋磺內酯鉀對於球體附著率無顯著影響。醋磺內酯鉀增加粒線體數量，並藉由增加CAT活性減少細胞內的ROS生成。另外，阿斯巴甜透過其代謝途徑生成細胞氧化壓力而提升CAT活性，沒有觀察到對於粒線體之影響。 Exposure of non-nutritive sweeteners during pregnancy increased ROS and reduced mitochondrial function, which affects the development of the placenta and fetus, thereby preventing implantation and pregnancy. The aim of this study was to investigate the effects of aspartame, sucralose and ACEK on endometrial cells and their attachment to trophoblast spheroids. Ishikawa cells were cultured for 24 hours with 0.5, 1 and 2 mM aspartame, sucralose and ACEK. Cellular or mitochondrial oxidative stress, mitochondrial membrane potential and mitochondrial mass were analyzed by fluorescent staining. Mitochondrial respiration oxygen consumption rate was analyzed by seahorse XFe-24 analyzer to evaluate mitochondrial function, antioxidant enzyme activity was analyzed by ELISA kit, and mitochondrial biogenesis related protein expression was analyzed by western blot. JEG-3 spheroids were co-cultured with Ishikawa cells, the spheroid attachment rate was evaluated as an in vitro implantation model. The results showed that sucralose increased ROS in cells and mitochondria, decreased mitochondrial SOD activity, inhibited mitochondrial respiration, increased mitochondrial membrane potential and compensatively increased mitochondrial number, so its treatment decreased spheroids adhesion rate. ACEK increased mitochondrial mass and reduced intracellular ROS production by increasing CAT activity. In addition, aspartame increased CAT activity through its metabolic pathway to generate cellular ROS, and no effect on mitochondria was observed.
描述:	碩士指導教授:謝榮鴻委員:葉添順委員:陳玉華委員:謝榮鴻
資料類型:	thesis
顯示於類別:	[保健營養學系暨研究所] 博碩士論文

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